Bioscience Reports
Portland Press Ltd.
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Anti-apoptosis mechanism of triptolide based on network pharmacology in focal segmental glomerulosclerosis rats
Volume: 40, Issue: 4
DOI 10.1042/BSR20192920
Abstract

Triptolide (TPL), the active component of Tripterygium wilfordii, exhibits anti-cancer and antioxidant functions. We aimed to explore the anti-apoptosis mechanism of TPL based on network pharmacology and in vivo and in vitro research validation using a rat model of focal segmental glomerulosclerosis (FSGS). The chemical structures and pharmacological activities of the compounds reported in T. wilfordii were determined and used to perform the network pharmacology analysis. The Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) was then used to identify the network targets for 16 compounds from Tripterygium wilfordii. Our results showed that 47 overlapping genes obtained from the GeneCards and OMIM databases were involved in the occurrence and development of FSGS and used to construct the protein–protein interaction (PPI) network using the STRING database. Hub genes were identified via the MCODE plug-in of the Cytoscape software. IL4 was the target gene of TPL in FSGS and was mainly enriched in the cell apoptosis term and p53 signaling pathway, according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. TPL inhibited FSGS-induced cell apoptosis in rats and regulated IL4, nephrin, podocin, and p53 protein levels via using CCK8, TUNEL, and Western blot assays. The effects of IL4 overexpression, including inhibition of cell viability and promotion of apoptosis, were reversed by TPL. TPL treatment increased the expression of nephrin and podocin and decreased p53 expression in rat podocytes. In conclusion, TPL inhibited podocyte apoptosis by targeting IL4 to alleviate kidney injury in FSGS rats.

Keywords
Li, Jiang, Song, Yang, and Pan: Anti-apoptosis mechanism of triptolide based on network pharmacology in focal segmental glomerulosclerosis rats

Introduction

Focal segmental glomerulosclerosis (FSGS) is a clinical pathological syndrome, and its typical pathological feature is sclerosing lesions in the focal glomeruli and in the glomerular segment. The clinical manifestations of FSGS patients are massive proteinuria, hematuria, hypertension, and progressive decrease in renal function. The condition of 3.6% of patients with end-stage renal disease developed from FSGS [1,2]. Currently, the main clinical therapies for FSGS are immunologic drugs, glucocorticoids, and blockers of the renin–angiotensin system; however, their therapeutic effects are not satisfactory. [3]. Triptolide (TPL) is the most active and effective diterpene lactone epoxide compound isolated from Tripterygium. [4]. TPL has anti-inflammatory, anti-tumor, and immunologic effects on many diseases [4]. TPL inhibits the secretion of many cytokines, adhesion molecules, and chemokines and affects the functions of various cells, including dendritic cells and renal tubular epithelial cells [5,6]. TPL has been reported to alleviate the progression of glomerulosclerosis and the excretion rate of urinary albumin to inhibit the progression of diabetic neuropathy [6]. However, the effects and mechanisms of TPL in FSGS are still unclear. We explored the mechanism of FSGS-mediated podocyte pathogenesis based on the FSGS rat model. The present study has great significance for the diagnosis, prevention, and treatment of FSGS.

In the past, research on Chinese herbal extracts focused on a particular aspect and on finding the biological characteristics explaining the pharmacological effect with respect to this aspect [7]; however, this approach is usually one-sided. It is important to explore the relation between the acquired proof and the research results. With the development of bioinformatics and network pharmacology, proposal of a theory and proving this theory through experiments has become the main method to explore the mechanism of Chinese herb compounds [8].

Network pharmacology is based on high-throughput omics data analysis, virtual computer computing and network database retrieval, and it combines systems biology with multidirectional pharmacology [9]. The mechanism of drug action was researched via the construction and analysis of biological networks. The systematic and holistic nature of network pharmacology is consistent with the characteristics of Chinese herbs, which exhibit multi-components, multi-targets, and systematic regulation. It has been widely used to explore the pharmacological basis of Chinese medicine and the drug mechanism and to interpret drug compatibility [10,11]. Network pharmacology has been recognized by many Chinese medicine researchers [12]. The multi-component and multi-target network research mode breaks the traditional research mode of a single ingredient and a single target, providing a new method for comprehensive analysis of the mechanism of the compounds [13]. In the early stage, a total of 47 target genes and the corresponding 16 active constituents of Tripterygium were used to construct the ingredient-target network. The present study mainly explores the mechanism of TPL in FSGS through bioinformatics and functional experiments.

Materials and methods

Construction of the potential compound database for tripterygium

Using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (http://lsp.nwu.edu.cn/tcmsp.php, TCMSP), each candidate’s drug ability was analyzed according to its oral bioavailability (OB) and drug-likeness (DL) indices recommended by the TCMSP. OB refers to the degree and rate of drug absorption into the circulatory system, which is an important indicator for objectively evaluating the intrinsic quality of drugs. The higher the OB of the compound, the more likely it is to be developed for clinical application. DL is the sum of the pharmacokinetic properties and safety, which arises from the interactions among the physicochemical properties and structural factors, including solubility, permeability, and stability. It can be used to optimize compounds, analyze the results of drug activity, predict in vivo pharmacokinetics, direct structure modifications, etc. As the TCMSP recommends, molecules with OB ≥ 30% and DL ≥ 0.18 were considered to exhibit relatively better pharmacological properties and were screened out as candidate compounds for further analysis.

Construction of the disease–target–compound network

To comprehensively understand the molecular mechanisms, disease–compound–target networks were constructed using the Cytoscape visualization software 3.7.1. All target genes related to FSGC were obtained from the GeneCards database (https://www.genecards.org/). All the candidate compounds of Tripterygium were retrieved from the TCMSP to obtain the associated targets. Next, disease, compounds, and targets were inputted into the software, and a disease–compound–target interaction network was constructed. In the process of constructing the network, the layout algorithm (attribute circle layout) was applied. We can set the geometric position of every node and visually display the network topology using color, graphics, and symbols, making reasonable arrangements for every node and creating a clear visual effect. Degree and betweenness centrality are two important parameters of the topology structure, which were used to evaluate the essentiality of each target and compound.

PPI network construction and module analysis

Search Tool for the Retrieval of Interacting Genes (STRING, https://string-db.org) is an online tool and used to construct the PPI network with confidence network edges and a medium confidence of 0.400 as the product criteria. Cytoscape 7.1.0 was used to perform the visualization of PPI network. The Molecular Complex Detection (MCODE) plug-in was used to screen the significant modules in the PPI network with a degree cut-off = 2, node score cut-off = 0.2, k-core = 2, and maximum depth = 100. The corresponding proteins in the central nodes and highly degree were potential core proteins encoded by key candidate genes that have important physiological regulatory functions.

GO terms and KEGG pathway enrichment analysis

The Database for Annotation, Visualization, and Integrated Discovery (DAVID) database was used to perform GO enrichment analysis and KEGG pathway enrichment analysis. The GO terms were classified into three categories: biological process (BP), cellular component (CC); and molecular function (MF). P<0.01 was considered to indicate a statistically significant difference.

Animals

A total of 40 Sprague Dawley (SD) rats (male, weighing 160–180 g) were provided by Yison Bio co.LTD (Shanghai, China). Animals were housed individually in polycarbonate cages with wood chip bedding and were maintained in an air-conditioned animal room (temperature: 24°C, relative humidity: 55 ± 5%) on a 12-h light/dark cycle. Each animal experiment was carried out following the local Care for Laboratory Animals guidelines formulated by the Animal Experimental Center. The Ethics Committee had approved the studies using laboratory animals at the Guangxing Affiliated Hospital of Zhejiang Chinese Medical University.

FSGS model establishment

All rats were randomly divided into a sham operation group (Sham), model group (FSGS), a group administered 80 mg/(kg·d) of Tripterygium by gavage (TPL(80)+FSGS), and a group administered 160 mg/(kg · d) of Tripterygium by gavage (TPL(160)+FSGS) (n=10 rats/group). One day before the operation, the TPL (80 or 160)+FSGS groups were administered TPL (Purifa Technology Development Co. Ltd., Chengdu, Sichuan, China) 80 or 160 mg/(kg d) by gavage; the Sham and FSGS groups were given isovolumic normal saline till the end of the experiment. The animals were intraperitoneally anesthetized with pentobarbital sodium (60 mg/kg body weight) and then placed on a homeothermic pad to maintain a core body temperature of 37°C to establish the FSGS model. The rats were first subjected to unilateral nephrectomy (left side) on day 1 and then injected in the caudal vein with adriamycin 5 mg/kg (on day 7) and adriamycin 3 mg/kg (on day 28) dissolved in 0.9% saline at a dilution of 2 mg/ml. Meanwhile, the kidneys of the control rats were exposed without dissecting the kidney tissue, followed by layer-by-layer suturing. These rats were then injected with saline on days 7 and 28 through the tail vein after the sham operation. Eight weeks post-surgery, blood samples were obtained from the tail veins, and the animals were killed. Following adequate anesthesia with pentobarbital sodium (180 mg/kg body weight), the organs were removed, frozen, or fixed in 4% paraformaldehyde. The serum and whole kidneys were harvested for biochemical, histological, and molecular analyses. The urinary protein levels of the rats were quantified before the end of the experiment. Animals with >100 mg/24 h urinary protein indicated successful establishment of the model, and they were included in subsequent experiments.

Histological analyses

The kidney tissues were fixed with 4% paraformaldehyde and embedded in paraffin. For histological analysis of lesions, 3 μm thick tissue sections were deparaffinized and stained with hematoxylin and eosin (HE) and periodic acid–Schiff (PAS). To calculate the degree of focal glomerular sclerosis, 40–60 glomeruli from each stained specimen were examined. The degree of sclerosis in each glomerulus was subjectively graded on a scale of 0–4 as follows: Grade 0, no change; Grade 1, sclerotic area less than or equal to 25% of the glomerulus or the presence of distinct adhesion between the capillary tuft and Bowman’s capsule; Grade 2, sclerosis of 25–50% of the total glomerular area; Grade 3, sclerosis of 50–75% of the total glomerular area; and Grade 4, sclerosis of more than 75% of the glomerulus. The glomerular sclerosis index (GSI) was calculated using the following formula:

GSI=(1×N1)+(2×N2)+(3×N3)+(4×N4)/(N0+N1+N2+N3+N4)
where N is the number of glomeruli for each grade of sclerosis.

Terminal dUTP nick-end labeling (TUNEL) staining

An apoptosis detection kit (Promega, Madison, WI) was used to detect apoptosis according to a previously described method [14]. In brief, renal sections were subjected to TUNEL staining in accordance with the manufacturer’s instructions. Later, IF microscopy was used to analyze the samples using a Zeiss Axiovert 200 M fluorescent microscope equipped with an AxioCamMR3 camera. Six fields (magnification 400×) were randomly selected from every section from 10 different rats, and cells with positive TUNEL staining were analyzed.

Glucose treatment and cell culture

Rat glomerular podocytes were provided by Yubo Bio-Technique Co. Ltd (Shanghai, China), which were then cultivated according to a previously described method. Rat podocytes were cultivated in RPMI 1640 (Sigma-Aldrich, U.S.A.) containing streptomycin (100 μg/ml), penicillin (100 U/ml) (Solarbio, Beijing, China), and 10% fetal bovine serum (FBS, Gibco, NY, Grand Island). Subsequently, the cells were cultivated in a 5% CO2 incubator (Heraeus, Japan) at 33°C with interferon-γ (IFN-γ, 40 units/ml, Sigma, St Louis, MO, U.S.A.). Later, to induce differentiation, the podocytes were maintained at 37°C for 2 weeks in the absence of interferon. Podocytes (3 × 105 cells/ml) were plated into 6-well plates in the presence of complete medium. After 24 h of standing, the podocytes were subjected to 24 and 48 h of TPL treatment at different concentrations (0, 5, 10, 20, 40, and 80 µmol/ml) before they were collected for subsequent analysis.

Transient transfection of plasmid DNA or siRNA

The previously described human IL4 plasmid DNA at full length [15] was utilized to increase IL4 expression in cells via using transient transfection. pcDNA3.1-Myc/His EV plasmid (Life technologies) and On-Target Plus scramble RNA (Dharmacon) were used as transient transfection controls. Sequences for IL4 overexpression was ACAUUACUGCCUGAAGGGUGAAUUAACGC.

Counting Kit-8 (CCK-8) assay

Cells were grown into the 96-well plates at the density of 1 × 105 cells/well, followed by 24 and 48 h of culture. Afterwards, cell viability was detected via using the CCK-8 kit (Dojindo Molecular Technologies, Gaithersburg, MD, U.S.A.). Then, cells in each group were cultivated for additional 24 and 48 h, respectively. Next, the CCK8 solution (10 μl) was added into cell at 37°C for 4 h. The absorbance was determined at 450 nm for obtaining the cell growth curve by the iMark microplate absorbance reader (Bio-Rad Laboratories, Inc., Hercules, CA, U.S.A.). Each experiment was carried out in triplicate.

Apoptosis detected by flow cytometry using Annexin V-FITC/PI staining

Cell apoptosis was examined using the Annexin V-FITC/PI kit. Briefly, the cells were subjected to 0.25% trypsin digestion (Thermo Fisher Scientific, Waltham, MA, U.S.A.), followed by two washes with cold PBS; resuspension with 5 μl of PI, 5 μl of annexin V-FITC, and 500 μl of binding buffer; and incubation under 15 min of ambient temperature in the dark. Typically, Annexin V-FITC can bind to phosphatidylserine located on the outer apoptotic cell membrane, whereas PI can penetrate and stain cells with impaired membranes before binding to and labeling DNA. Data were collected using a flow cytometer (BD FACSCalibur; BD Biosciences, Franklin Lakes, NJ, U.S.A.) and analyzed by FlowJo. Clumped cells were excluded from the FSC-H/FSC-A dot plot for selecting the single cells. Cells in the annexin V-FITC-/PI+, annexin V-FITC+/PI+, and annexin V-FITC+/PI− quadrants were regarded as apoptotic cells.

Western blotting

Cells were subjected to lysis within the RIPA buffer (Beyotime, Shanghai, China) to collect the lysates in tubes, followed by 20 min of centrifugation at 4°C at 12,000 g. Later, all supernatants were extracted, and protein content was measured by the BCA Protein Quantitative Kit (Beyotime, Shanghai, China). Then, Western blotting had been carried out in accordance with the instruction. Afterwards, 20 μg protein was subjected to 10% SDS-PAGE for separation, followed by transfer onto the PVDF membranes (Millipore, Billerica, MA, U.S.A.). Later, the membranes were blocked using 5% skimmed milk for 1 h, followed by overnight incubation with anti-IL-4 (dilution 1:8000, Abcam, Cambridge, MA, U.S.A., ab69811), nephrin (Abcam, ab227806; diluted at 1:1200), podocin (Abcam, ab50339; diluted at 1:1000), phosph (p)-Stat6 (BioVision, U.S.A., 3476-100; diluted at 1:1000), and GAPDH (Abcam, Cambridge, MA, U.S.A., ab181602; dilution 1:1000) rabbit anti-human antibodies, at 4°C, separately. Afterwards, cells were subjected to 1 h incubation with HRP-labeled secondary antibody (goat anti-rabbit antibody, Abcam, Cambridge, MA, U.S.A., ab116282; dilution 1:2000), prior to ECL detection. The Immobilon Western Chemiluminescent kit (WBKLS0100; Millipore, U.S.A.) was used to reveal the reactive bands using Roche Cobas e601 automated chemiluminescence image analysis system (Roche, U.S.A.).

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay

The GAPDH and IL-4 mRNA expression was detected through RT-qPCR. Total cellular RNA was isolated by TRIzol (Invitrogen, Carlsbad, CA, U.S.A.) in accordance with manufacturer protocols. Afterwards, cDNA was synthesized by reverse transcription of RNA according to the reactions below: RNase-free dH2O, total RNA (500 ng), and 5×PrimeScript RT Master Mix (2 μl) were added until the final volume became 10 μl. The Prism 7500 (ABI, Foster City, CA, U.S.A.) was employed for real-time PCR following the standard protocol of SYBR green assay. Primers used in the present study were shown below: IL-4-F: 5′-GATCACAAAGTACTGGTCCTGG-3′. Notably, GAPDH served as a normal control, with the primers of 5′-CACCCTGTTGCTGTAGCCAAA-3′ (reverse) and 5′-TGACTTCAACAGCGACACCCA-3′ (forward). Later, qPCR was carried out in triplicate using 7500 Real-Time PCR ABI system (ABI, U.S.A.) at a format of the 96-well plate. The reaction volume of 20 μl was prepared for PCR, which included forward primer (0.8 μl, 10 μM), RNase-free dH2O (7.4 μl),reverse primer (0.8 μl, 10 μM), 2×FastStart Universal SYBR Green Master (10 μl, ROX; Invitrogen, Guangzhou, China), and cDNA (1 μl). Besides, the PCR conditions were as follows, 10 min at 95°C, followed by 15 s at 95°C for 40 cycles, and 1 min at 60°C. The sequence detection software (1.6.3, Applied Biosystems, ABI, U.S.A.) was used for data analysis. Relative GAPDH or IL-4 mRNA level was measured and standardized according to 2−ΔΔCt method based on GAPDH.

Statistical analyses

SPSS 15.0 (http://spss.en.softonic.com/) was employed for all statistical analyses. Differences between two groups were analyzed through independent sample t-test, whereas those among several groups were examined by one-way analysis of variance (ANOVA). Rate was compared by chi-square test. The statistically significant level was set as P<0.01 or P<0.05.

Results

Identification of active compounds in Tripterygium wilfordii

Using TCMSP databases (http://lsp.nwsuaf.edu.cn/tcmsp.php), 144 compounds of Tripterygium were retrieved. According to the criteria of DL ≥ 0.18 and OB ≥ 30%, a total of 51 chemical ingredients were selected (Table 1). TPL was verified as an active ingredient of T. wilfordii.

Table 1
Information for 51 chemical ingredients of tripterygium
Mol IDMolecule nameOB (%)DL
MOL000211Mairin55.380.78
MOL000296Hederagenin36.910.75
MOL000358Beta-sitosterol36.910.75
MOL000422Kaempferol41.880.24
MOL000449Stigmasterol43.830.76
MOL00205840957-99-157.20.62
MOL003182(+)-Medioresinol di-O-beta-D-glucopyranoside_qt60.690.62
MOL00318481827-74-945.420.53
MOL003185(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a
-dimethyl-4,9,10,10a-tetrahydro-3H-phenanthren-2-one
48.840.38
MOL003187Triptolide51.290.68
MOL003188Tripchlorolide78.720.72
MOL003189WILFORLIDE A35.660.72
MOL003192Triptonide67.660.7
MOL003196Tryptophenolide48.50.44
MOL0031985 alpha-Benzoyl-4 alpha-hydroxy-1 beta,8 alpha-dinicotinoyl-dihydro-agarofuran35.260.72
MOL0031995,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin61.850.54
MOL003206Canin77.410.33
MOL003208Celafurine72.940.44
MOL003209Celallocinnine83.470.59
MOL003210Celapanine30.180.82
MOL003211Celaxanthin47.370.58
MOL003217Isoxanthohumol56.810.39
MOL003222Salazinic acid36.340.76
MOL003224Tripdiotolnide56.40.67
MOL003225Hypodiolide A76.130.49
MOL003229Triptinin B34.730.32
MOL003231Triptoditerpenic acid B40.020.36
MOL003232Triptofordin B139.550.84
MOL003233Triptofordin B2107.710.76
MOL003234Triptofordin C230.160.76
MOL003235Triptofordin D1320.75
MOL003236Triptofordin D230.380.69
MOL003238Triptofordin F133.910.6
MOL003239Triptofordin F233.620.67
MOL003241Triptofordin F431.370.67
MOL003242Triptofordinine A230.780.47
MOL003244Triptonide68.450.68
MOL003245Triptonoditerpenic acid42.560.39
MOL003248Triptonoterpene48.570.28
MOL00326621-Hydroxy-30-norhopan-22-one34.110.77
MOL003267Wilformine46.320.2
MOL003278Salaspermic acid32.190.63
MOL00327999694-86-775.230.66
MOL003280TRIPTONOLIDE49.510.49
MOL003283(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol66.510.39
MOL004443Zhebeiresinol58.720.19
MOL005828Nobiletin61.670.52
MOL007415[(2S)-2-[[(2S)-2-(benzoylamino)-3-phenylpropanoyl]amino]-3-phenylpropyl] acetate58.020.52
MOL007535(5S,8S,9S,10R,13R,14S,17R)-17-[(1R,4R)-4-ethyl-1,5-dimethylhexyl]
-10,13-dimethyl-2,4,5,7,8,9,11,12,14,15,16,
17-dodecahydro-1H-cyclopenta
[a]phenanthrene-3,6-dione
33.120.79
MOL0093863,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran52.110.54
MOL011169Peroxyergosterol44.390.82

Construction of the disease–target–compound network and PPI network

The TCMSP and GeneCards databases were used to predict the potential targets for each compound in FSGS. As a result, 123 target genes from the GeneCards database were verified to be involved in FSGS (Table 2), and 695 target genes of Tripterygium from the TCMSP database were verified (Table 3). After importing data into Cytoscape, a disease–compound–target network was constructed (Figure 1A). In addition, 47 overlapping target genes from two databases (TCMSP and GeneCards) were used to construct the PPI network. IL4 obtained from the most significant module of the PPI network was verified as a key by using the MCODE plug-in of Cytoscape software, and it was found to be involved in FSGS (Figure 1B–D).

Disease–Compound–Target network and PPI network
Figure 1
(A) The triptergium-target network of FSGS. (B) 47 overlapped target genes were from two databases (TCMSP and GeneCards). (C) 47 overlapped target genes were used to constructed PPI network and hub genes in PPI network. (D) Module and key gene were analysis and screened by using the MCODE plug-in Cytoscape software.Disease–Compound–Target network and PPI network
Table 2
Information for target genes of FSGS from GeneCards database
Gene symbolDescription
INF2Inverted Formin, FH2 And WH2 Domain Containing
TRPC6Transient Receptor Potential Cation Channel Subfamily C Member 6
CD2APCD2 Associated Protein
ACTN4Actinin Alpha 4
NPHS1NPHS1, Nephrin
NPHS2NPHS2, Podocin
PAX2Paired Box 2
WT1Wilms Tumor 1
CRB2Crumbs 2, Cell Polarity Complex Component
MYO1EMyosin IE
APOL1Apolipoprotein L1
ANLNAnillin Actin Binding Protein
PLCE1Phospholipase C Epsilon 1
PTPROProtein Tyrosine Phosphatase, Receptor Type O
NUP107Nucleoporin 107
ARHGAP24Rho GTPase Activating Protein 24
SMARCAL1SWI/SNF Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A Like 1
COQ6Coenzyme Q6, Monooxygenase
LAMB2Laminin Subunit Beta 2
COL4A3Collagen Type IV Alpha 3 Chain
NUP93Nucleoporin 93
COQ8BCoenzyme Q8B
SYNPOSynaptopodin
TGFB1Transforming Growth Factor Beta 1
CLCN5Chloride Voltage-Gated Channel 5
MYH9Myosin Heavy Chain 9
LOC107985291Uncharacterized LOC107985291
COL4A4Collagen Type IV Alpha 4 Chain
SGPL1Sphingosine-1-Phosphate Lyase 1
NUP205Nucleoporin 205
PLAURPlasminogen Activator, Urokinase Receptor
ACEAngiotensin I Converting Enzyme
COL4A5Collagen Type IV Alpha 5 Chain
CDKN1CCyclin Dependent Kinase Inhibitor 1C
KIRREL2Kirre Like Nephrin Family Adhesion Molecule 2
CDKN1ACyclin Dependent Kinase Inhibitor 1A
EMP2Epithelial Membrane Protein 2
CDKN1BCyclin Dependent Kinase Inhibitor 1B
ILKIntegrin Linked Kinase
CD80CD80 Molecule
SLC37A4Solute Carrier Family 37 Member 4
CTNNB1Catenin Beta 1
ITGA3Integrin Subunit Alpha 3
SCARB2Scavenger Receptor Class B Member 2
AGRNAgrin
ALG1ALG1, Chitobiosyldiphosphodolichol Beta-Mannosyltransferase
PLEKHH2Pleckstrin Homology, MyTH4 And FERM Domain Containing H2
MPV17Mitochondrial Inner Membrane Protein MPV17
ALBAlbumin
WDR73WD Repeat Domain 73
RENRenin
NARS2Asparaginyl-TRNA Synthetase 2, Mitochondrial
SEC61A1Sec61 Translocon Alpha 1 Subunit
ZNF592Zinc Finger Protein 592
ITGB4Integrin Subunit Beta 4
IL1BInterleukin 1 Beta
OSGEPO-Sialoglycoprotein Endopeptidase
TP53RKTP53 Regulating Kinase
TPRKBTP53RK Binding Protein
LAGE3L Antigen Family Member 3
G6PCGlucose-6-Phosphatase Catalytic Subunit
VPS33AVPS33A, CORVET/HOPS Core Subunit
LPLLipoprotein Lipase
ICAM1Intercellular Adhesion Molecule 1
CDH17Cadherin 17
MAGI2Membrane Associated Guanylate Kinase, WW And PDZ Domain Containing 2
ARHGDIARho GDP Dissociation Inhibitor Alpha
DNAI1Dynein Axonemal Intermediate Chain 1
ANKFY1Ankyrin Repeat And FYVE Domain Containing 1
BSNDBarttin CLCNK Type Accessory Beta Subunit
MAXMYC Associated Factor X
FAHFumarylacetoacetate Hydrolase
VHLVon Hippel–Lindau Tumor Suppressor
LYZLysozyme
AFPAlpha Fetoprotein
RETRet Proto-Oncogene
MDH2Malate Dehydrogenase 2
SDHBSuccinate Dehydrogenase Complex Iron Sulfur Subunit B
SDHASuccinate Dehydrogenase Complex Flavoprotein Subunit A
MUC1Mucin 1, Cell Surface Associated
FHFumarate Hydratase
KIF1BKinesin Family Member 1B
SDHCSuccinate Dehydrogenase Complex Subunit C
SDHDSuccinate Dehydrogenase Complex Subunit D
COQ2Coenzyme Q2, Polyprenyltransferase
PLECPlectin
SDHAF2Succinate Dehydrogenase Complex Assembly Factor 2
TMEM127Transmembrane Protein 127
ELP1Elongator Complex Protein 1
ACHEAcetylcholinesterase (Cartwright Blood Group)
TJP1Tight Junction Protein 1
KIRREL1Kirre Like Nephrin Family Adhesion Molecule 1
NEDENephropathy, Progressive, With Deafness
PDGFAPlatelet Derived Growth Factor Subunit A
CD40LGCD40 Ligand
ENTPD5Ectonucleoside Triphosphate Diphosphohydrolase 5
GAPVD1GTPase Activating Protein And VPS9 Domains 1
NUMBLNUMB Like, Endocytic Adaptor Protein
KANK2KN Motif And Ankyrin Repeat Domains 2
CD79ACD79a Molecule
BRAFB-Raf Proto-Oncogene, Serine/Threonine Kinase
NDNNecdin, MAGE Family Member
AXDND1Axonemal Dynein Light Chain Domain Containing 1
PLCE1-AS1PLCE1 Antisense RNA 1
LMX1BLIM Homeobox Transcription Factor 1 Beta
WT1-ASWT1 Antisense RNA
PODXLPodocalyxin Like
INSInsulin
VIMVimentin
NAGLUN-Acetyl-Alpha-Glucosaminidase
ACTBActin Beta
NR5A1Nuclear Receptor Subfamily 5 Group A Member 1
LOC105369403Uncharacterized LOC105369403
ITGB1Integrin Subunit Beta 1
UTRNUtrophin
ALG13ALG13, UDP-N-Acetylglucosaminyltransferase Subunit
CLDN1Claudin 1
CCN2Cellular Communication Network Factor 2
OCRLOCRL, Inositol Polyphosphate-5-Phosphatase
CMIPC-Maf Inducing Protein
ACTL7BActin Like 7B
NXF5Nuclear RNA Export Factor 5
CUBNCubilin
LCATLecithin-Cholesterol Acyltransferase
AGTR1Angiotensin II Receptor Type 1
LRP2LDL Receptor Related Protein 2
TNFTumor Necrosis Factor
BAXBCL2 Associated X, Apoptosis Regulator
TLR4Toll Like Receptor 4
ITGB3Integrin Subunit Beta 3
DAG1Dystroglycan 1
CCL2C-C Motif Chemokine Ligand 2
NGFNerve Growth Factor
CYCSCytochrome C, Somatic
VTNVitronectin
SMAD3SMAD Family Member 3
NOTCH1Notch 1
WNT1Wnt Family Member 1
CCNA2Cyclin A2
NTRK2Neurotrophic Receptor Tyrosine Kinase 2
DBHDopamine Beta-Hydroxylase
SMARCA4SWI/SNF Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A, Member 4
SMARCA2SWI/SNF Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A, Member 2
IDSIduronate 2-Sulfatase
PYGLGlycogen Phosphorylase L
GUSBGlucuronidase Beta
GALK1Galactokinase 1
APRTAdenine Phosphoribosyltransferase
ALADAminolevulinate Dehydratase
PAX6Paired Box 6
SOX9SRY-Box 9
CLCN7Chloride Voltage-Gated Channel 7
ALDOBAldolase, Fructose-Bisphosphate B
HYAL1Hyaluronidase 1
PHKA2Phosphorylase Kinase Regulatory Subunit Alpha 2
HEXAHexosaminidase Subunit Alpha
HPD4-Hydroxyphenylpyruvate Dioxygenase
ARSBArylsulfatase B
APOC2Apolipoprotein C2
GALNSGalactosamine (N-Acetyl)-6-Sulfatase
CLCNKBChloride Voltage-Gated Channel Kb
AMHAnti-Mullerian Hormone
GNSGlucosamine (N-Acetyl)-6-Sulfatase
SGSHN-Sulfoglucosamine Sulfohydrolase
FGF9Fibroblast Growth Factor 9
CLCN4Chloride Voltage-Gated Channel 4
IDUAIduronidase, Alpha-L-
TIA1TIA1 Cytotoxic Granule Associated RNA Binding Protein
INPP5BInositol Polyphosphate-5-Phosphatase B
CLCNKAChloride Voltage-Gated Channel Ka
CLDN16Claudin 16
G6PC3Glucose-6-Phosphatase Catalytic Subunit 3
BAZ1ABromodomain Adjacent To Zinc Finger Domain 1A
GSTZ1Glutathione S-Transferase Zeta 1
CIAO1Cytosolic Iron-Sulfur Assembly Component 1
ELP3Elongator Acetyltransferase Complex Subunit 3
SMARCA1SWI/SNF Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A, Member 1
GABREGamma-Aminobutyric Acid Type A Receptor Epsilon Subunit
USP19Ubiquitin Specific Peptidase 19
ELP2Elongator Acetyltransferase Complex Subunit 2
TIMM8BTranslocase Of Inner Mitochondrial Membrane 8 Homolog B
CPSF7Cleavage And Polyadenylation Specific Factor 7
ASTN1Astrotactin 1
LHX9LIM Homeobox 9
SRYSex Determining Region Y
ZNF274Zinc Finger Protein 274
ARSHArylsulfatase Family Member H
MFRPMembrane Frizzled-Related Protein
TECPR2Tectonin Beta-Propeller Repeat Containing 2
YIPF3Yip1 Domain Family Member 3
ZFYZinc Finger Protein Y-Linked
FAM47EFamily With Sequence Similarity 47 Member E
LOC100506321Uncharacterized LOC100506321
LAMB1Laminin Subunit Beta 1
PCNAProliferating Cell Nuclear Antigen
MT-ND2Mitochondrially Encoded NADH:Ubiquinone Oxidoreductase Core Subunit 2
MT-CO1Mitochondrially Encoded Cytochrome C Oxidase I
MT-CO2Mitochondrially Encoded Cytochrome C Oxidase II
CTSLCathepsin L
SERPINE1Serpin Family E Member 1
EZREzrin
IFI27Interferon Alpha Inducible Protein 27
AKT1AKT Serine/Threonine Kinase 1
CCND1Cyclin D1
BMP6Bone Morphogenetic Protein 6
LMNALamin A/C
CR1Complement C3b/C4b Receptor 1 (Knops Blood Group)
SMAD2SMAD Family Member 2
AGTAngiotensinogen
CLUClusterin
CCNB1Cyclin B1
PPARGPeroxisome Proliferator Activated Receptor Gamma
TIMP2TIMP Metallopeptidase Inhibitor 2
IL6Interleukin 6
EDN1Endothelin 1
DNM1Dynamin 1
CDH2Cadherin 2
JAG1Jagged 1
MMEMembrane Metalloendopeptidase
CAMK2BCalcium/Calmodulin Dependent Protein Kinase II Beta
FYNFYN Proto-Oncogene, Src Family Tyrosine Kinase
LRP5LDL Receptor Related Protein 5
PTK2Protein Tyrosine Kinase 2
LRP6LDL Receptor Related Protein 6
VCLVinculin
ITGAVIntegrin Subunit Alpha V
KRT8Keratin 8
PLCG1Phospholipase C Gamma 1
DKK1Dickkopf WNT Signaling Pathway Inhibitor 1
CD151CD151 Molecule (Raph Blood Group)
NCK1NCK Adaptor Protein 1
YWHAQTyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein Theta
IRF6Interferon Regulatory Factor 6
PARVAParvin Alpha
KIRREL3Kirre Like Nephrin Family Adhesion Molecule 3
MKI67Marker Of Proliferation Ki-67
LAMA5Laminin Subunit Alpha 5
TLN1Talin 1
LIMS1LIM Zinc Finger Domain Containing 1
FAT1FAT Atypical Cadherin 1
MIR4758MicroRNA 4758
MIR6852MicroRNA 6852
IL2Interleukin 2
PON1Paraoxonase 1
FN1Fibronectin 1
IL2RAInterleukin 2 Receptor Subunit Alpha
IL10Interleukin 10
NOS2Nitric Oxide Synthase 2
CABIN1Calcineurin Binding Protein 1
FGF2Fibroblast Growth Factor 2
LCN2Lipocalin 2
LAMC1Laminin Subunit Gamma 1
CDK2Cyclin Dependent Kinase 2
APOEApolipoprotein E
PLA2G7Phospholipase A2 Group VII
HIF1AHypoxia Inducible Factor 1 Subunit Alpha
PAFAH1B1Platelet Activating Factor Acetylhydrolase 1b Regulatory Subunit 1
F2RCoagulation Factor II Thrombin Receptor
GNA12G Protein Subunit Alpha 12
TTRTransthyretin
MMP14Matrix Metallopeptidase 14
ACTN1Actinin Alpha 1
ATP7AATPase Copper Transporting Alpha
IGFBP3Insulin Like Growth Factor Binding Protein 3
ATP6AP2ATPase H+ Transporting Accessory Protein 2
GNEGlucosamine (UDP-N-Acetyl)-2-Epimerase/N-Acetylmannosamine Kinase
S100A4S100 Calcium Binding Protein A4
ENPEPGlutamyl Aminopeptidase
ZMPSTE24Zinc Metallopeptidase STE24
AMBPAlpha-1-Microglobulin/Bikunin Precursor
NPNTNephronectin
CDK4Cyclin Dependent Kinase 4
PLAUPlasminogen Activator, Urokinase
RARARetinoic Acid Receptor Alpha
MTHFRMethylenetetrahydrofolate Reductase
VLDLRVery Low Density Lipoprotein Receptor
CYP11B2Cytochrome P450 Family 11 Subfamily B Member 2
EYA1EYA Transcriptional Coactivator And Phosphatase 1
GPX3Glutathione Peroxidase 3
LTBP1Latent Transforming Growth Factor Beta Binding Protein 1
IGFBP1Insulin Like Growth Factor Binding Protein 1
PTPRUProtein Tyrosine Phosphatase, Receptor Type U
MAGI1Membrane Associated Guanylate Kinase, WW And PDZ Domain Containing 1
RAP1GAPRAP1 GTPase Activating Protein
NPHP4Nephrocystin 4
PDGFBPlatelet Derived Growth Factor Subunit B
SLC12A1Solute Carrier Family 12 Member 1
FBXW7F-Box And WD Repeat Domain Containing 7
FABP1Fatty Acid Binding Protein 1
THBDThrombomodulin
CLCF1Cardiotrophin Like Cytokine Factor 1
CHKACholine Kinase Alpha
IFNA2Interferon Alpha 2
ECT2Epithelial Cell Transforming 2
COG2Component Of Oligomeric Golgi Complex 2
PDSS2Decaprenyl Diphosphate Synthase Subunit 2
FMN1Formin 1
SDK1Sidekick Cell Adhesion Molecule 1
MIR186MicroRNA 186
MIR193AMicroRNA 193a
MTORMechanistic Target Of Rapamycin Kinase
HMGCR3-Hydroxy-3-Methylglutaryl-CoA Reductase
MMP2Matrix Metallopeptidase 2
TGFBR1Transforming Growth Factor Beta Receptor 1
A2MAlpha-2-Macroglobulin
TFAMTranscription Factor A, Mitochondrial
NRF1Nuclear Respiratory Factor 1
IGFBP2Insulin Like Growth Factor Binding Protein 2
SMAD1SMAD Family Member 1
IGF1RInsulin Like Growth Factor 1 Receptor
IGF1Insulin Like Growth Factor 1
IRS1Insulin Receptor Substrate 1
SRCSRC Proto-Oncogene, Non-Receptor Tyrosine Kinase
SLC2A1Solute Carrier Family 2 Member 1
APOC1Apolipoprotein C1
GAPDHGlyceraldehyde-3-Phosphate Dehydrogenase
GIPRGastric Inhibitory Polypeptide Receptor
F2RL3F2R Like Thrombin Or Trypsin Receptor 3
DGKQDiacylglycerol Kinase Theta
VEGFAVascular Endothelial Growth Factor A
TIMP1TIMP Metallopeptidase Inhibitor 1
RHOARas Homolog Family Member A
MIFMacrophage Migration Inhibitory Factor
IL4Interleukin 4
MAPK14Mitogen-Activated Protein Kinase 14
DDIT3DNA Damage Inducible Transcript 3
RBP4Retinol Binding Protein 4
SP1Sp1 Transcription Factor
FOSFos Proto-Oncogene, AP-1 Transcription Factor Subunit
LDLRLow Density Lipoprotein Receptor
TNFSF11TNF Superfamily Member 11
SOD1Superoxide Dismutase 1
TTC21BTetratricopeptide Repeat Domain 21B
RAC1Rac Family Small GTPase 1
ANGPTL4Angiopoietin Like 4
SMAD7SMAD Family Member 7
MAPK1Mitogen-Activated Protein Kinase 1
MPOMyeloperoxidase
ACE2Angiotensin I Converting Enzyme 2
MYCMYC Proto-Oncogene, BHLH Transcription Factor
ABCB1ATP Binding Cassette Subfamily B Member 1
HGFHepatocyte Growth Factor
B2MBeta-2-Microglobulin
MAPK3Mitogen-Activated Protein Kinase 3
ENGEndoglin
PPARAPeroxisome Proliferator Activated Receptor Alpha
BCL2BCL2, Apoptosis Regulator
HMOX1Heme Oxygenase 1
CCL5C-C Motif Chemokine Ligand 5
IL15Interleukin 15
HPXHemopexin
ESR1Estrogen Receptor 1
EGFEpidermal Growth Factor
CASP3Caspase 3
NR3C1Nuclear Receptor Subfamily 3 Group C Member 1
NRP1Neuropilin 1
TNFRSF11ATNF Receptor Superfamily Member 11a
CD2CD2 Molecule
GREM1Gremlin 1, DAN Family BMP Antagonist
MIR30AMicroRNA 30a
CXCR4C-X-C Motif Chemokine Receptor 4
JAK3Janus Kinase 3
TLR3Toll Like Receptor 3
FTH1Ferritin Heavy Chain 1
NOTCH2Notch 2
SIRT1Sirtuin 1
EPAS1Endothelial PAS Domain Protein 1
GGT1Gamma-Glutamyltransferase 1
ABCA1ATP Binding Cassette Subfamily A Member 1
CASP9Caspase 9
NFATC1Nuclear Factor Of Activated T Cells 1
YAP1Yes Associated Protein 1
GFERGrowth Factor, Augmenter Of Liver Regeneration
CEBPACCAAT Enhancer Binding Protein Alpha
LIPCLipase C, Hepatic Type
HSP90B1Heat Shock Protein 90 Beta Family Member 1
SMAD6SMAD Family Member 6
ATF3Activating Transcription Factor 3
PROM1Prominin 1
AGTR2Angiotensin II Receptor Type 2
LGALS1Galectin 1
NRP2Neuropilin 2
SP3Sp3 Transcription Factor
DDNDendrin
CD24CD24 Molecule
MIR30DMicroRNA 30d
METMET Proto-Oncogene, Receptor Tyrosine Kinase
PRKCDProtein Kinase C Delta
CTSDCathepsin D
CASP8Caspase 8
FASFas Cell Surface Death Receptor
TFTransferrin
ALOX5Arachidonate 5-Lipoxygenase
KRT18Keratin 18
RELARELA Proto-Oncogene, NF-KB Subunit
BDNFBrain Derived Neurotrophic Factor
CTLA4Cytotoxic T-Lymphocyte Associated Protein 4
LTA4HLeukotriene A4 Hydrolase
NLRP3NLR Family Pyrin Domain Containing 3
HSPA5Heat Shock Protein Family A (Hsp70) Member 5
HSPG2Heparan Sulfate Proteoglycan 2
CXCL12C-X-C Motif Chemokine Ligand 12
SPP1Secreted Phosphoprotein 1
TRPV5Transient Receptor Potential Cation Channel Subfamily V Member 5
COL4A6Collagen Type IV Alpha 6 Chain
PDGFDPlatelet Derived Growth Factor D
IL13Interleukin 13
IL9Interleukin 9
HBEGFHeparin Binding EGF Like Growth Factor
LTC4SLeukotriene C4 Synthase
TRAF1TNF Receptor Associated Factor 1
WWC1WW And C2 Domain Containing 1
VASPVasodilator Stimulated Phosphoprotein
EPOErythropoietin
HHIPHedgehog Interacting Protein
GNPTABN-Acetylglucosamine-1-Phosphate Transferase Subunits Alpha And Beta
ADAM19ADAM Metallopeptidase Domain 19
CAPZA1Capping Actin Protein Of Muscle Z-Line Subunit Alpha 1
ATL1Atlastin GTPase 1
PFN2Profilin 2
PDLIM1PDZ And LIM Domain 1
STK16Serine/Threonine Kinase 16
IL7Interleukin 7
TRPC1Transient Receptor Potential Cation Channel Subfamily C Member 1
SNX9Sorting Nexin 9
UBDUbiquitin D
EPB41L5Erythrocyte Membrane Protein Band 4.1 Like 5
PDLIM2PDZ And LIM Domain 2
ETV7ETS Variant 7
ACTL7AActin Like 7A
MIR10AMicroRNA 10a
MIR135A1MicroRNA 135a-1
MIR135BMicroRNA 135b
MIR217MicroRNA 217
MIR378AMicroRNA 378a
MIR135A2MicroRNA 135a-2
MT-TL1Mitochondrially Encoded TRNA Leucine 1 (UUA/G)
HNP1Hypertensive Nephropathy
AGERAdvanced Glycosylation End-Product Specific Receptor
GLAGalactosidase Alpha
CXCL8C-X-C Motif Chemokine Ligand 8
AKR1B1Aldo-Keto Reductase Family 1 Member B
JUNJun Proto-Oncogene, AP-1 Transcription Factor Subunit
NOS3Nitric Oxide Synthase 3
COL4A2Collagen Type IV Alpha 2 Chain
KNG1Kininogen 1
MMP9Matrix Metallopeptidase 9
TGFBR2Transforming Growth Factor Beta Receptor 2
DESDesmin
PRKCBProtein Kinase C Beta
DCNDecorin
VCAM1Vascular Cell Adhesion Molecule 1
HRASHRas Proto-Oncogene, GTPase
CASP1Caspase 1
IFNGR1Interferon Gamma Receptor 1
NR1H2Nuclear Receptor Subfamily 1 Group H Member 2
CFBComplement Factor B
ANTXR2ANTXR Cell Adhesion Molecule 2
MSR1Macrophage Scavenger Receptor 1
CASP4Caspase 4
HLA-DRB1Major Histocompatibility Complex, Class II, DR Beta 1
IL12AInterleukin 12A
COX5ACytochrome C Oxidase Subunit 5A
HPHaptoglobin
PRTN3Proteinase 3
OLR1Oxidized Low Density Lipoprotein Receptor 1
HLA-DQB1Major Histocompatibility Complex, Class II, DQ Beta 1
EREGEpiregulin
DIAPH2Diaphanous Related Formin 2
AZGP1Alpha-2-Glycoprotein 1, Zinc-Binding
AREGAmphiregulin
PTAFRPlatelet Activating Factor Receptor
TLE4Transducin Like Enhancer Of Split 4
IL12BInterleukin 12B
BPIBactericidal Permeability Increasing Protein
SCGB1A1Secretoglobin Family 1A Member 1
IFNA1Interferon Alpha 1
SEMA4CSemaphorin 4C
ADCK2AarF Domain Containing Kinase 2
MIR196A2MicroRNA 196a-2
MIR490MicroRNA 490
SMAD4SMAD Family Member 4
EDNRAEndothelin Receptor Type A
PLATPlasminogen Activator, Tissue Type
E2F1E2F Transcription Factor 1
ITIH4Inter-Alpha-Trypsin Inhibitor Heavy Chain Family Member 4
MIR21MicroRNA 21
MMP1Matrix Metallopeptidase 1
CATCatalase
MAPK10Mitogen-Activated Protein Kinase 10
PARP1Poly(ADP-Ribose) Polymerase 1
RB1RB Transcriptional Corepressor 1
ESR2Estrogen Receptor 2
CD36CD36 Molecule
GDNFGlial Cell Derived Neurotrophic Factor
LEPLeptin
NPPANatriuretic Peptide A
MBL2Mannose Binding Lectin 2
CST3Cystatin C
SEMA3ASemaphorin 3A
THBS1Thrombospondin 1
UMODUromodulin
SERPINB7Serpin Family B Member 7
AKT2AKT Serine/Threonine Kinase 2
NFKB1Nuclear Factor Kappa B Subunit 1
STAT3Signal Transducer And Activator Of Transcription 3
CDC42Cell Division Cycle 42
CYP3A4Cytochrome P450 Family 3 Subfamily A Member 4
NFKBIANFKB Inhibitor Alpha
MAPK8Mitogen-Activated Protein Kinase 8
CD40CD40 Molecule
C3Complement C3
PLGPlasminogen
MMP7Matrix Metallopeptidase 7
PTK2BProtein Tyrosine Kinase 2 Beta
DDX58DExD/H-Box Helicase 58
COL4A1Collagen Type IV Alpha 1 Chain
PIK3CGPhosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Gamma
MYH7Myosin Heavy Chain 7
IGF2Insulin Like Growth Factor 2
ECE1Endothelin Converting Enzyme 1
C5Complement C5
COL1A2Collagen Type I Alpha 2 Chain
PROS1Protein S
MYH6Myosin Heavy Chain 6
TNCTenascin C
VCANVersican
GFPT1Glutamine–Fructose-6-Phosphate Transaminase 1
EDN3Endothelin 3
CCR1C-C Motif Chemokine Receptor 1
ADIPOQAdiponectin, C1Q And Collagen Domain Containing
TRIOTrio Rho Guanine Nucleotide Exchange Factor
FABP4Fatty Acid Binding Protein 4
CCR2C-C Motif Chemokine Receptor 2
CSF1Colony Stimulating Factor 1
BMP7Bone Morphogenetic Protein 7
S100A8S100 Calcium Binding Protein A8
MLXIPLMLX Interacting Protein Like
TNFRSF12ATNF Receptor Superfamily Member 12A
PLTPPhospholipid Transfer Protein
PDPNPodoplanin
NID1Nidogen 1
FMODFibromodulin
NESNestin
SLC25A17Solute Carrier Family 25 Member 17
TNFSF12TNF Superfamily Member 12
USF2Upstream Transcription Factor 2, C-Fos Interacting
ZFYVE9Zinc Finger FYVE-Type Containing 9
PITRM1Pitrilysin Metallopeptidase 1
SMPDL3BSphingomyelin Phosphodiesterase Acid Like 3B
CCN1Cellular Communication Network Factor 1
PDGFRAPlatelet Derived Growth Factor Receptor Alpha
EGFREpidermal Growth Factor Receptor
PDGFRBPlatelet Derived Growth Factor Receptor Beta
JAK2Janus Kinase 2
KDRKinase Insert Domain Receptor
MAP2K1Mitogen-Activated Protein Kinase Kinase 1
MAP2K2Mitogen-Activated Protein Kinase Kinase 2
INSRInsulin Receptor
ARAndrogen Receptor
AKT3AKT Serine/Threonine Kinase 3
PIK3CAPhosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha
PTENPhosphatase And Tensin Homolog
STAT1Signal Transducer And Activator Of Transcription 1
CDK5Cyclin Dependent Kinase 5
ADKAdenosine Kinase
MMP3Matrix Metallopeptidase 3
UCHL1Ubiquitin C-Terminal Hydrolase L1
CD4CD4 Molecule
CDKN2ACyclin Dependent Kinase Inhibitor 2A
CTSBCathepsin B
CASP7Caspase 7
ACVRL1Activin A Receptor Like Type 1
ADAM17ADAM Metallopeptidase Domain 17
PPIBPeptidylprolyl Isomerase B
HSPB1Heat Shock Protein Family B (Small) Member 1
YWHAETyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein Epsilon
GATA3GATA Binding Protein 3
JAK1Janus Kinase 1
HDAC3Histone Deacetylase 3
NTRK1Neurotrophic Receptor Tyrosine Kinase 1
NOS1Nitric Oxide Synthase 1
CCNE1Cyclin E1
ACACAAcetyl-CoA Carboxylase Alpha
CAV1Caveolin 1
PRKCZProtein Kinase C Zeta
SGK1Serum/Glucocorticoid Regulated Kinase 1
NR3C2Nuclear Receptor Subfamily 3 Group C Member 2
SLC5A1Solute Carrier Family 5 Member 1
TFRCTransferrin Receptor
TGFB2Transforming Growth Factor Beta 2
VWFVon Willebrand Factor
FOXO1Forkhead Box O1
CYP3A5Cytochrome P450 Family 3 Subfamily A Member 5
FASLGFas Ligand
CCR5C-C Motif Chemokine Receptor 5 (Gene/Pseudogene)
CES1Carboxylesterase 1
CNR1Cannabinoid Receptor 1
PPARDPeroxisome Proliferator Activated Receptor Delta
SELPSelectin P
MMP10Matrix Metallopeptidase 10
NR1H3Nuclear Receptor Subfamily 1 Group H Member 3
MS4A1Membrane Spanning 4-Domains A1
SPHK1Sphingosine Kinase 1
LRP1LDL Receptor Related Protein 1
HDAC9Histone Deacetylase 9
TGFATransforming Growth Factor Alpha
TRAF3TNF Receptor Associated Factor 3
TRAF6TNF Receptor Associated Factor 6
TSHRThyroid Stimulating Hormone Receptor
ADORA2BAdenosine A2b Receptor
BIDBH3 Interacting Domain Death Agonist
CA8Carbonic Anhydrase 8
CDK5R1Cyclin Dependent Kinase 5 Regulatory Subunit 1
CFHComplement Factor H
CDK1Cyclin Dependent Kinase 1
ANXA5Annexin A5
ANGAngiogenin
ITGB6Integrin Subunit Beta 6
LNPEPLeucyl And Cystinyl Aminopeptidase
SERPINH1Serpin Family H Member 1
S100BS100 Calcium Binding Protein B
TBX3T-Box 3
PXNPaxillin
TGIF1TGFB Induced Factor Homeobox 1
TNFSF13BTNF Superfamily Member 13b
ZYXZyxin
GH1Growth Hormone 1
CX3CR1C-X3-C Motif Chemokine Receptor 1
LGALS3Galectin 3
HIPK2Homeodomain Interacting Protein Kinase 2
STMN1Stathmin 1
HPSEHeparanase
EGR1Early Growth Response 1
CD34CD34 Molecule
EEF1A1Eukaryotic Translation Elongation Factor 1 Alpha 1
CTNSCystinosin, Lysosomal Cystine Transporter
BDKRB2Bradykinin Receptor B2
HDAC7Histone Deacetylase 7
IQGAP1IQ Motif Containing GTPase Activating Protein 1
SALL1Spalt Like Transcription Factor 1
MPZMyelin Protein Zero
MEFVMEFV, Pyrin Innate Immunity Regulator
TATTyrosine Aminotransferase
SPI1Spi-1 Proto-Oncogene
RAB3ARAB3A, Member RAS Oncogene Family
USF1Upstream Transcription Factor 1
FOXO3Forkhead Box O3
DDAH2Dimethylarginine Dimethylaminohydrolase 2
FCGR3BFc Fragment Of IgG Receptor IIIb
IL17AInterleukin 17A
P2RX4Purinergic Receptor P2X 4
PLXNA1Plexin A1
TGThyroglobulin
TNFRSF6BTNF Receptor Superfamily Member 6b
CSRP3Cysteine And Glycine Rich Protein 3
ACTC1Actin, Alpha, Cardiac Muscle 1
C4AComplement C4A (Rodgers Blood Group)
TAGLNTransgelin
ID1Inhibitor Of DNA Binding 1, HLH Protein
CCL4C-C Motif Chemokine Ligand 4
FCARFc Fragment Of IgA Receptor
CAMPCathelicidin Antimicrobial Peptide
COL8A2Collagen Type VIII Alpha 2 Chain
ST3GAL4ST3 Beta-Galactoside Alpha-2,3-Sialyltransferase 4
IL1RL1Interleukin 1 Receptor Like 1
WASLWiskott-Aldrich Syndrome Like
CHIAChitinase, Acidic
BPHLBiphenyl Hydrolase Like
KLF15Kruppel Like Factor 15
PLA2R1Phospholipase A2 Receptor 1
RHODRas Homolog Family Member D
SCAPSREBF Chaperone
PDLIM5PDZ And LIM Domain 5
RPH3ARabphilin 3A
HIST1H1BHistone Cluster 1 H1 Family Member B
CCL3C-C Motif Chemokine Ligand 3
COL8A1Collagen Type VIII Alpha 1 Chain
PECAM1Platelet And Endothelial Cell Adhesion Molecule 1
P3H1Prolyl 3-Hydroxylase 1
NUP133Nucleoporin 133
SNF8SNF8, ESCRT-II Complex Subunit
TSLPThymic Stromal Lymphopoietin
ACTN3Actinin Alpha 3 (Gene/Pseudogene)
LECT2Leukocyte Cell Derived Chemotaxin 2
WDR19WD Repeat Domain 19
CSN1S1Casein Alpha S1
GOLIM4Golgi Integral Membrane Protein 4
MPV17LMPV17 Mitochondrial Inner Membrane Protein Like
WTIPWT1 Interacting Protein
HIST2H3CHistone Cluster 2 H3 Family Member C
KRBOX4KRAB Box Domain Containing 4
MIR216AMicroRNA 216a
MBL3PMannose-Binding Lectin Family Member 3, Pseudogene
Table 3
Information for target genes of tripterygium from TCMSP database
Mol IdGene NameMol name
MOL000296PGRhederagenin
MOL000296NCOA2hederagenin
MOL000296CHRM3hederagenin
MOL000296CHRM1hederagenin
MOL000296CHRM2hederagenin
MOL000296ADRA1Bhederagenin
MOL000296GABRA1hederagenin
MOL000296GRIA2hederagenin
MOL000296ADH1Bhederagenin
MOL000296ADH1Chederagenin
MOL000296LYZhederagenin
MOL000296PTGS1hederagenin
MOL000296SCN5Ahederagenin
MOL000296PTGS2hederagenin
MOL000296RXRAhederagenin
MOL000296SLC6A2hederagenin
MOL003182KCNH2(+)-Medioresinol di-O-beta-D-glucopyranoside_qt
MOL003182SCN5A(+)-Medioresinol di-O-beta-D-glucopyranoside_qt
MOL003182PTGS2(+)-Medioresinol di-O-beta-D-glucopyranoside_qt
MOL003182F7(+)-Medioresinol di-O-beta-D-glucopyranoside_qt
MOL003184PTGS181827-74-9
MOL003184CHRM381827-74-9
MOL003184KCNH281827-74-9
MOL003184CHRM181827-74-9
MOL003184SCN5A81827-74-9
MOL003184CHRM581827-74-9
MOL003184PTGS281827-74-9
MOL003184CHRM481827-74-9
MOL003184OPRD181827-74-9
MOL003184PGR81827-74-9
MOL003184CHRM281827-74-9
MOL003184ADRA1B81827-74-9
MOL003184ADRB281827-74-9
MOL003184OPRM181827-74-9
MOL003184NCOA281827-74-9
MOL003184NCOA181827-74-9
MOL003185CHRM3(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a
-dimethyl-4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185CHRM1(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl
-4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185PTGS2(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl
-4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185OPRD1(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl
-4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185ADRA1A(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185ADRA1B(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185ADRA1D(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185OPRM1(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185NR3C1(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185NCOA2(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185NCOA1(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185SCN5A(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185CHRM2(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003185ADRB2(1R,4aR,10aS)-5-hydroxy-1-(hydroxymethyl)-7-isopropyl-8-methoxy-1,4a-dimethyl-
4,9,10,10a-tetrahydro-3H-phenanthren-2-one
MOL003187RELAtriptolide
MOL003187STAT3triptolide
MOL003187VEGFAtriptolide
MOL003187BCL2triptolide
MOL003187FOStriptolide
MOL003187CDKN1Atriptolide
MOL003187PLAUtriptolide
MOL003187TNFSF15triptolide
MOL003187JUNtriptolide
MOL003187CASP3triptolide
MOL003187TP63triptolide
MOL003187MAPK8triptolide
MOL003187PTGS2triptolide
MOL003187STAT1triptolide
MOL003187CXCL8triptolide
MOL003187MCL1triptolide
MOL003187IL2triptolide
MOL003187IFNGtriptolide
MOL003187IL4triptolide
MOL003187CD80triptolide
MOL003187CD86triptolide
MOL003187CXCR4triptolide
MOL003187BIRC3triptolide
MOL003187CD274triptolide
MOL003187IL23Atriptolide
MOL003187CCR7triptolide
MOL003187CD1Atriptolide
MOL003187CD40triptolide
MOL003187CD14triptolide
MOL003187C3triptolide
MOL003187VTCN1triptolide
MOL003196CHRM3Tryptophenolide
MOL003196KCNH2Tryptophenolide
MOL003196CHRM1Tryptophenolide
MOL003196SCN5ATryptophenolide
MOL003196CHRM5Tryptophenolide
MOL003196PTGS2Tryptophenolide
MOL003196RXRATryptophenolide
MOL003196OPRD1Tryptophenolide
MOL003196ADRA1ATryptophenolide
MOL003196PGRTryptophenolide
MOL003196CHRM2Tryptophenolide
MOL003196ADRA1BTryptophenolide
MOL003196ADRB2Tryptophenolide
MOL003196ADRA1DTryptophenolide
MOL003196OPRM1Tryptophenolide
MOL003196NCOA2Tryptophenolide
MOL003196NCOA1Tryptophenolide
MOL003199NOS25,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199PTGS15,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199KCNH25,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199ESR15,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199AR5,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199SCN5A5,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199PPARG5,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199PTGS25,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199F75,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199KDR5,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199PYGM5,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003199PRSS15,8-Dihydroxy-7-(4-hydroxy-5-methyl-coumarin-3)-coumarin
MOL003209KCNH2Celallocinnine
MOL003209SCN5ACelallocinnine
MOL003217NOS2Isoxanthohumol
MOL003217KCNH2Isoxanthohumol
MOL003217ESR1Isoxanthohumol
MOL003217SCN5AIsoxanthohumol
MOL003217PTGS2Isoxanthohumol
MOL003217KDRIsoxanthohumol
MOL003217ADRA1BIsoxanthohumol
MOL003217ADRB2Isoxanthohumol
MOL003217NCOA2Isoxanthohumol
MOL003217NCOA1Isoxanthohumol
MOL003217PTGS1Isoxanthohumol
MOL003217PPARDIsoxanthohumol
MOL003224NR3C2Tripdiotolnide
MOL003225NR3C2Hypodiolide A
MOL003225NR3C1Hypodiolide A
MOL003229CHRM3Triptinin B
MOL003229KCNH2Triptinin B
MOL003229CHRM1Triptinin B
MOL003229SCN5ATriptinin B
MOL003229CHRM5Triptinin B
MOL003229PTGS2Triptinin B
MOL003229RXRATriptinin B
MOL003229ADRA1ATriptinin B
MOL003229PGRTriptinin B
MOL003229CHRM2Triptinin B
MOL003229ADRA1BTriptinin B
MOL003229ADRB2Triptinin B
MOL003229ADRA1DTriptinin B
MOL003229OPRM1Triptinin B
MOL003229NR3C1Triptinin B
MOL003229RXRBTriptinin B
MOL003229NCOA2Triptinin B
MOL003229NCOA1Triptinin B
MOL003231PTGS1Triptoditerpenic acid B
MOL003231CHRM3Triptoditerpenic acid B
MOL003231KCNH2Triptoditerpenic acid B
MOL003231CHRM1Triptoditerpenic acid B
MOL003231SCN5ATriptoditerpenic acid B
MOL003231CHRM5Triptoditerpenic acid B
MOL003231PTGS2Triptoditerpenic acid B
MOL003231CHRM4Triptoditerpenic acid B
MOL003231RXRATriptoditerpenic acid B
MOL003231OPRD1Triptoditerpenic acid B
MOL003231ADRA1ATriptoditerpenic acid B
MOL003231PGRTriptoditerpenic acid B
MOL003231CHRM2Triptoditerpenic acid B
MOL003231ADRA1BTriptoditerpenic acid B
MOL003231ADRB2Triptoditerpenic acid B
MOL003231ADRA1DTriptoditerpenic acid B
MOL003231OPRM1Triptoditerpenic acid B
MOL003231NR3C1Triptoditerpenic acid B
MOL003231RXRBTriptoditerpenic acid B
MOL003231NCOA2Triptoditerpenic acid B
MOL003231NCOA1Triptoditerpenic acid B
MOL003245CHRM3Triptonoditerpenic acid
MOL003245KCNH2Triptonoditerpenic acid
MOL003245CHRM1Triptonoditerpenic acid
MOL003245SCN5ATriptonoditerpenic acid
MOL003245PTGS2Triptonoditerpenic acid
MOL003245OPRD1Triptonoditerpenic acid
MOL003245ADRA1BTriptonoditerpenic acid
MOL003245ADRB2Triptonoditerpenic acid
MOL003245NCOA2Triptonoditerpenic acid
MOL003245NCOA1Triptonoditerpenic acid
MOL003248PTGS1Triptonoterpene
MOL003248CHRM3Triptonoterpene
MOL003248CHRM1Triptonoterpene
MOL003248SCN5ATriptonoterpene
MOL003248PTGS2Triptonoterpene
MOL003248RXRATriptonoterpene
MOL003248ACHETriptonoterpene
MOL003248ADRA1ATriptonoterpene
MOL003248PGRTriptonoterpene
MOL003248CHRM2Triptonoterpene
MOL003248ADRA1BTriptonoterpene
MOL003248ADRB2Triptonoterpene
MOL003248ADRA1DTriptonoterpene
MOL003248OPRM1Triptonoterpene
MOL003248NR3C1Triptonoterpene
MOL003248NCOA2Triptonoterpene
MOL003248NCOA1Triptonoterpene
MOL003266PGR21-Hydroxy-30-norhopan-22-one
MOL003280CHRM3TRIPTONOLIDE
MOL003280CHRM1TRIPTONOLIDE
MOL003280SCN5ATRIPTONOLIDE
MOL003280CHRM5TRIPTONOLIDE
MOL003280PTGS2TRIPTONOLIDE
MOL003280OPRD1TRIPTONOLIDE
MOL003280ADRA1ATRIPTONOLIDE
MOL003280PGRTRIPTONOLIDE
MOL003280CHRM2TRIPTONOLIDE
MOL003280ADRB2TRIPTONOLIDE
MOL003280OPRM1TRIPTONOLIDE
MOL003280NCOA2TRIPTONOLIDE
MOL003280NCOA1TRIPTONOLIDE
MOL000358PGRbeta-sitosterol
MOL000358NCOA2beta-sitosterol
MOL000358PTGS1beta-sitosterol
MOL000358PTGS2beta-sitosterol
MOL000358KCNH2beta-sitosterol
MOL000358CHRM3beta-sitosterol
MOL000358CHRM1beta-sitosterol
MOL000358SCN5Abeta-sitosterol
MOL000358CHRM4beta-sitosterol
MOL000358ADRA1Abeta-sitosterol
MOL000358CHRM2beta-sitosterol
MOL000358ADRA1Bbeta-sitosterol
MOL000358ADRB2beta-sitosterol
MOL000358CHRNA2beta-sitosterol
MOL000358SLC6A4beta-sitosterol
MOL000358OPRM1beta-sitosterol
MOL000358GABRA1beta-sitosterol
MOL000358BCL2beta-sitosterol
MOL000358BAXbeta-sitosterol
MOL000358CASP9beta-sitosterol
MOL000358JUNbeta-sitosterol
MOL000358CASP3beta-sitosterol
MOL000358CASP8beta-sitosterol
MOL000358PRKCAbeta-sitosterol
MOL000358PON1beta-sitosterol
MOL000358MAP2beta-sitosterol
MOL000211PGRMairin
MOL000422NOS2kaempferol
MOL000422PTGS1kaempferol
MOL000422ARkaempferol
MOL000422PPARGkaempferol
MOL000422PTGS2kaempferol
MOL000422NCOA2kaempferol
MOL000422PRSS1kaempferol
MOL000422PGRkaempferol
MOL000422CHRM1kaempferol
MOL000422ACHEkaempferol
MOL000422SLC6A2kaempferol
MOL000422CHRM2kaempferol
MOL000422ADRA1Bkaempferol
MOL000422GABRA1kaempferol
MOL000422F7kaempferol
MOL000422RELAkaempferol
MOL000422IKBKBkaempferol
MOL000422AKT1kaempferol
MOL000422BCL2kaempferol
MOL000422BAXkaempferol
MOL000422TNFSF15kaempferol
MOL000422JUNkaempferol
MOL000422AHSA1kaempferol
MOL000422CASP3kaempferol
MOL000422MAPK8kaempferol
MOL000422MMP1kaempferol
MOL000422STAT1kaempferol
MOL000422PPARGkaempferol
MOL000422HMOX1kaempferol
MOL000422CYP3A4kaempferol
MOL000422CYP1A2kaempferol
MOL000422CYP1A1kaempferol
MOL000422ICAM1kaempferol
MOL000422SELEkaempferol
MOL000422VCAM1kaempferol
MOL000422NR1I2kaempferol
MOL000422CYP1B1kaempferol
MOL000422ALOX5kaempferol
MOL000422HAS2kaempferol
MOL000422GSTP1kaempferol
MOL000422AHRkaempferol
MOL000422PSMD3kaempferol
MOL000422SLC2A4kaempferol
MOL000422NR1I3kaempferol
MOL000422INSRkaempferol
MOL000422DIO1kaempferol
MOL000422PPP3CAkaempferol
MOL000422GSTM1kaempferol
MOL000422GSTM2kaempferol
MOL000422AKR1C3kaempferol
MOL000422SLPIkaempferol
MOL000449PGRStigmasterol
MOL000449NR3C2Stigmasterol
MOL000449NCOA2Stigmasterol
MOL000449ADH1CStigmasterol
MOL000449RXRAStigmasterol
MOL000449NCOA1Stigmasterol
MOL000449PTGS1Stigmasterol
MOL000449PTGS2Stigmasterol
MOL000449ADRA2AStigmasterol
MOL000449SLC6A2Stigmasterol
MOL000449SLC6A3Stigmasterol
MOL000449ADRB2Stigmasterol
MOL000449AKR1B1Stigmasterol
MOL000449PLAUStigmasterol
MOL000449LTA4HStigmasterol
MOL000449MAOBStigmasterol
MOL000449MAOAStigmasterol
MOL000449CTRB1Stigmasterol
MOL000449CHRM3Stigmasterol
MOL000449CHRM1Stigmasterol
MOL000449ADRB1Stigmasterol
MOL000449SCN5AStigmasterol
MOL000449ADRA1AStigmasterol
MOL000449CHRM2Stigmasterol
MOL000449ADRA1BStigmasterol
MOL000449GABRA1Stigmasterol
MOL002058KCNH240957-99-1
MOL002058SCN5A40957-99-1
MOL002058PTGS240957-99-1
MOL002058PTGS140957-99-1
MOL002058NCOA240957-99-1
MOL002058F740957-99-1
MOL003283ESR1(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283AR(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283PPARG(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283PTGS2(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283F7(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283ADRB2(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283ESR2(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283MAPK14(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283GSK3B(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283CHEK1(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283NCOA2(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283SCN5A(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283CCNA2(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL003283PTGS1(2R,3R,4S)-4-(4-hydroxy-3-methoxy-phenyl)-7-methoxy-2,3-dimethylol-tetralin-6-ol
MOL004443PTGS1Zhebeiresinol
MOL004443SCN5AZhebeiresinol
MOL004443PTGS2Zhebeiresinol
MOL004443RXRAZhebeiresinol
MOL004443ADRB2Zhebeiresinol
MOL004443GABRA1Zhebeiresinol
MOL005828NOS2nobiletin
MOL005828PTGS1nobiletin
MOL005828KCNH2nobiletin
MOL005828ESR1nobiletin
MOL005828ARnobiletin
MOL005828PPARGnobiletin
MOL005828PTGS2nobiletin
MOL005828F7nobiletin
MOL005828ESR2nobiletin
MOL005828CHEK1nobiletin
MOL005828PRSS1nobiletin
MOL005828NCOA2nobiletin
MOL005828GSK3Bnobiletin
MOL005828SCN5Anobiletin
MOL005828BCL2nobiletin
MOL005828BAXnobiletin
MOL005828CASP9nobiletin
MOL005828MMP9nobiletin
MOL005828JUNnobiletin
MOL005828TP63nobiletin
MOL005828MAPK8nobiletin
MOL005828TIMP1nobiletin
MOL005828PPARGnobiletin
MOL005828CREB1nobiletin
MOL005828PLA2G4Anobiletin
MOL005828CD163nobiletin
MOL005828EPHB2nobiletin
MOL007415KCNH2[(2S)-2-[[(2S)-2-(benzoylamino)-3-phenylpropanoyl]amino]-3-phenylpropyl] acetate
MOL007415PTGS2[(2S)-2-[[(2S)-2-(benzoylamino)-3-phenylpropanoyl]amino]-3-phenylpropyl] acetate
MOL007415PRSS1[(2S)-2-[[(2S)-2-(benzoylamino)-3-phenylpropanoyl]amino]-3-phenylpropyl] acetate
MOL007535PGR(5S,8S,9S,10R,13R,14S,17R)-17-[(1R,4R)-4-ethyl-1,5-dimethylhexyl]-10,13-dimethyl-2,4,5,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene-3,6-dione
MOL009386KCNH23,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran
MOL009386ESR13,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran
MOL009386PTGS23,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran
MOL009386ADRB23,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran
MOL009386CCNA23,3′-bis-(3,4-dihydro-4-hydroxy-6-methoxy)-2H-1-benzopyran

Enrichment analysis of GO and KEGG

Forty-seven overlapping targets were screened for further investigation using the DAVID (htTPLs://david.ncifcrf.gov/) online tool. GO annotation results showed that the top 30 biological processes (BP) included cell apoptosis, proliferation of cells, positive signal transduction regulation, extracellular stimulus response, and cell death (Figure 2A). The results of KEGG enrichment analysis showed that the 47 overlapping targets were markedly enriched within 32 pathways, including the p53 signal transduction pathway, apoptosis, and the JAK-STAT signal transduction pathway (Figure 2B). IL4 was mainly enriched in BP terms, including programmed cell death regulation, endogenous stimulus response, apoptosis regulation, positive cell proliferation regulation, and positive nitrogen compound metabolic process regulation. Based on the KEGG enrichment results, IL4 participated in the T-cell receptor signal transduction pathway, allograft rejection, intestinal IgA production immunologic network, the JAK-STAT signal transduction pathway, autoimmune thyroid disease, the Fc epsilon RI signal transduction pathway, and the interaction between cytokines and cytokine receptors.

Analysis of GO and KEGG Enrichment
Figure 2
(A) 47 overlapped genes were analysis by GO annotation, which showed that the top 30 biological processes (BP). (B) 47 overlapped genes was analysis by KEGG, which enriched in 32 pathways.Analysis of GO and KEGG Enrichment

TPL alleviated kidney injury by inhibiting cell apoptosis in FSGS rats, and IL4 was up-regulated in kidney tissues of FSGS rats

FSGS rat models were established using external jugular vein cannulation; subsequently, the levels of BUN, 24-h urine protein, Scr, ALB, and TC were determined. Our results showed that the BUN, 24-h urine protein, TC, and Scr levels in FSGS animals were evidently decreased, while the ALB levels were significantly increased after the FSGS rats were administered TPL gavage (at 80 or 160 μg/(kg·d)) (Figure 3A–E). HE staining results showed that TPL significantly decreased the glomerulosclerosis index (GSI) in FSGS rats (Figure 3F,G). The apoptosis level was determined by TUNEL assay in the kidney tissues of FSGS rats. We found that FSGS promoted apoptosis in kidney tissues. However, TPL treatment suppressed the apoptosis of cells within the renal tissues of FSGS rats (Figure 4A,B). Therefore, we further detected the protein levels of IL4, nephrin, and podocin and the phosphorylation level of Stat6 using Western blotting. According to our results, TPL treatment decreased IL4 protein levels and stat6 activation, and increased the protein levels of nephrin and podocin in FSGS rats (Figure 4C–G).

TPL alleviated kidney injure in FSGS rats
Figure 3
FSGS rat models were establised by using external jugular vein cannulation, (A) 24 h urine protein, (B) BUN, (C) Scr, (D) TC and (E) ALB levels were detected; (F) TPL could significantly decrease glomerulosclerosis index (GSI) of FSGS rats; (G) The pathomorphology of kidney in FSGS rats was showed by HE staining. Data are presented as the mean ± standard deviation. aP<0.05 versus Sham group, bP<0.05 versus FSGS group, and cP<0.05 versus TPL(80)+FSGS group.TPL alleviated kidney injure in FSGS rats
TPL reduced cell apoptosis in FSGS rats
Figure 4
FSGS rat models were established by using external jugular vein cannulation. (A and B) Apoptosis level was determined by TUNEL assay in kidney tissues of FSGS rats. (C) The protein levels of IL4, nephrin and podocin, and phosphorylation level of Stat6 by Western blotting analysis. (D–G) Histogram showed the statistical value. GAPDH was used as a load control. Data are presented as the mean ± standard deviation. aP<0.05 versus Sham group, bP<0.05 versus FSGS group, and cP<0.05 versus TPL(80)+FSGS group.TPL reduced cell apoptosis in FSGS rats

TPL reversed the function of IL4 overexpression, promoting cell apoptosis

According to the results, 0–80 μmol/ml TPL had no influence on cell viability and apoptosis (Figure 5A–C). Western blotting results showed that 0–80 μmol/ml TPL minimally affected IL4, nephrin, and podocin expression and stat6 activation (Figure 5D). However, IL4 overexpression inhibited the viability and promoted apoptosis of podocytes. TPL inhibited IL4 overexpression-mediated cell apoptosis (Figure 6A–C). Furthermore, TPL decreased IL4 protein levels, increased nephrin and podocin protein levels, and inhibited the phosphorylation of Stat6 in podocytes (Figure 6D).

TPL has no influence on the cell viability and apoptosis
Figure 5
(A) 0–80 μmol/ml of TPL little effected the viability of podocytes by CCK8 assay. (B and C) 0–80 μmol/ml of TPL little affected the apoptosis level of podocytes by flow cytometry assay. (D) The protein levels of IL4, nephrin and podocin, and phosphorylation level of Stat6 by Western blotting analysis. GAPDH was used as a load control. Data are presented as the mean ± standard deviation. aP<0.05 versus Sham group, bP<0.05 versus FSGS group, and cP<0.05 versus TPL(80)+FSGS group.TPL has no influence on the cell viability and apoptosis
TPL reversed the function of IL4 overexpression promoting cell apoptosis
Figure 6
(A) IL4 protein and mRNA levels were detected by Western blot and RT-PCR assays. (B) The viability of podocytes by CCK8 assay in cell with IL4. (C) The apoptosis level of podocytes by flow cytometry assay in cell with IL4. (D) The protein levels of nephrin and podocin, and phosphorylation level of Stat6 by Western blotting analysis. GAPDH was used as a load control. Data are presented as the mean ± standard deviation. aP<0.05 versus vector group, and bP<0.05 versus IL4 group.TPL reversed the function of IL4 overexpression promoting cell apoptosis

Discussion

The occurrence of FSGS is related to a variety of mechanisms. Podocyte injury is the central link of FSGS [16,17]. Glomerular sclerosis is the final pathological change in FSGS caused by the excessive accumulation of the glomerular extracellular matrix (ECM). Podocytes are an important part of the glomerulus and are the final barriers that block the filtration of plasma macromolecules. Apoptosis, fusion, and shedding of podocytes induced the occurrence and development of FSGS. TPL has been reported to have a protective effect on kidney damage [18]. Therefore, we constructed the Disease–Target–Compound network in the present study through the TCM network pharmacology to confirm the relationship between TPL and FSGS. It was further confirmed by constructing a PPT network that IL4 was a target gene of TPL and FSGS. According to KEGG and GO enrichment analyses, IL4 was closely related to apoptosis and was enriched in the JAK-STAT signal pathway. Thus, we proposed two hypotheses: (1) TPL can protect against FSGS kidney injury by inhibiting apoptosis; (2) The protective effect of TPL on FSGS-induced kidney damage may be achieved by targeting IL4.

IL-4 is an anti-inflammatory factor that belongs to the interleukin family [19]. It has been reported that IL4 can inhibit apoptosis of liver cancer cells, and blockage of the IL4/IL4R/STAT6 axis can promote apoptosis of Hodgkin lymphoma cells [20]. However, IL4 may also be involved in the disease as a pro-inflammatory factor [21]. The expression level of IL4 is high in kidney tissue with acute kidney injury [22]. Therefore, the effect of IL4 on FSGS should be more extensively investigated. IL4 activates stat6 by acting on the JAK-STAT signal pathway. Our results demonstrated that IL4 expression and the phosphorylation level of stat6 were up-regulated in kidney tissues of FSGS rats. This suggests that the IL/STAT6 signaling pathway is aberrantly activated in FSGS. TPL reduced apoptosis in the kidney tissue of FSGS rats while significantly inhibiting the expression of IL4 and the activation of stat6.

Nephrin and podocin are podocyte proteins that have been widely used to identify kidney injury [23,24]. It has been reported that podocin and nephrin levels were down-regulated in a kidney injury model to promote podocyte apoptosis, thereby aggravating kidney damage. Podocin and nephrin expression levels were remarkably down-regulated in the kidney tissue of FSGS rats. Similarly, TPL could upgrade the podocin and nephrin expression levels. This indicated that TPL attenuated glomerular sclerosis in FSGS rats by reducing podocyte apoptosis. To further investigate the mechanism of action of TPL on renal protection in FSGS rats, we carried out a study at the cellular level.

First, we need to investigate if 0–80 µmol/ml of TPL is toxic to podocyte. The functional experiment proved that TPL at low concentrations did not affect cell activity; cell apoptosis; the expression of IL4, nephrin, and podocin; and the activation of stat6, which excluded the threat of TPL for cells. The results showed that a high expression of IL4 inhibited cell viability, promoted apoptosis, increased phosphorylation of stat3, and inhibited the expression of nephrin and podocin. This suggested that a high expression of IL4 promoted apoptosis and aggravated glomerular sclerosis. TPL can reverse IL4-mediated podocyte apoptosis and reduce glomerular sclerosis.

In conclusion, up-regulation of IL4 in kidney tissue of FSGS rats activated stat6 and promoted podocyte apoptosis to aggravate glomerular sclerosis. TPL can alleviate glomerular sclerosis in FSGS rats by inhibiting the activation of the IL4/stat6 signaling pathway and podocyte apoptosis. This finding can offer a theoretical foundation for the application of TPL in treating FSGS.

Competing Interests

The authors declare that there are no competing interests associated with the manuscript.

Funding

The research was funded by National Natural Science Foundation of China [grant number 81673913].

Author Contribution

Yayu Li and Xue Jiang wrote the main manuscript and analyzed the data. Yayu Li, Xue Jiang and Litao Song performed the experiments. Yayu Li, Mengdie Yang and Jing Pan designed the study. All authors read and approved the final manuscript.

Data Availability

The datasets used and/or analyzed during the present study are available from the corresponding author on reasonable request.

Abbreviations

FSGS

focal segmental glomerulosclerosis

GO

Gene Ontology

KEGG

Kyoto Encyclopedia of Genes and Genomes

PPI

protein–protein interaction

TCMSP

Traditional Chinese Medicine Systems Pharmacology Database

TPL

triptolide

References

1. 

Zhuo L., Huang L., Yang Z., Li G. and Wang L. (2019) . A comprehensive analysis of NPHS1 gene mutations in patients with sporadic focal segmental glomerulosclerosis. BMC Med. Genet.20, , pp.111, doi: 10.1186/s12881-019-0845-4

2. 

Snoek R., Nguyen T.Q., van der Zwaag B., van Zuilen A.D., Kruis H.M.E., van Gils-Verrij L.A.et al. (2019) . Importance of Genetic Diagnostics in Adult-Onset Focal Segmental Glomerulosclerosis. Nephron142, , pp.351–358, doi: 10.1159/000499937

3. 

Louis M., Cottenet J., Salmon-Rousseau A., Blot M., Bonnot P.H., Rebibou J.M.et al. (2019) . Prevalence and incidence of kidney diseases leading to hospital admission in people living with HIV in France: an observational nationwide study. BMJ Open9, , pp.e029211, doi: 10.1136/bmjopen-2019-029211

4. 

Feng Y., Zheng C., Zhang Y., Xing C., Cai W., Li R.et al. (2019) . Triptolide Inhibits Preformed Fibril-Induced Microglial Activation by Targeting the MicroRNA155-5p/SHIP1 Pathway. Oxidative Med. Cell. Longev.2019, , pp.6527638, doi: 10.1155/2019/6527638

5. 

Zhang G., Chen J., Liu Y., Yang R., Guo H. and Sun Z. (2013) . Triptolide-conditioned dendritic cells induce allospecific T-cell regulation and prolong renal graft survival. J. Invest. Surg.: Off. J. Acad. Surg. Res.26, , pp.191–199, doi: 10.3109/08941939.2012.737408

6. 

Xue M., Cheng Y., Han F., Chang Y., Yang Y., Li X.et al. (2018) . Triptolide Attenuates Renal Tubular Epithelial-mesenchymal Transition Via the MiR-188-5p-mediated PI3K/AKT Pathway in Diabetic Kidney Disease. Int. J. of Biol. Sci.14, , pp.1545–1557, doi: 10.7150/ijbs.24032

7. 

Deng B., Deng C. and Cheng Z. (2017) . Chinese Herbal Extractions for Relieving Radiation Induced Lung Injury: A Systematic Review and Meta-Analysis. Evidence-based Complement. Altern. Med.: eCAM2017, , pp.2141645, doi: 10.1155/2017/2141645

8. 

Wang Y.Y., Bai H., Zhang R.Z., Yan H., Ning K. and Zhao X.M. (2017) . Predicting new indications of compounds with a network pharmacology approach: Liuwei Dihuang Wan as a case study. Oncotarget8, , pp.93957–93968

9. 

Chen G., Huang C., Liu Y., Chen T., Huang R., Liang M.et al. (2018) . A Network Pharmacology Approach to Uncover the Potential Mechanism of Yinchensini Decoction. Evidence-based Complement. Altern. Med.: eCAM2018, , pp.2178610, doi: 10.1155/2018/2178610

10. 

Yin S.H., Xu P., Wang B., Lu Y., Wu Q.Y., Zhou M.L.et al. (2019) . Duration of dual antiplatelet therapy after percutaneous coronary intervention with drug-eluting stent: systematic review and network meta-analysis. BMJ365, , pp.l2222

11. 

Zhang W.N., Yang L., He S.S., Qin X.M. and Li A.P. (2019) . Metabolomics coupled with integrative pharmacology reveal the protective effect of FangjiHuangqi Decoction against adriamycin-induced rat nephropathy model. J. Pharm. Biomed. Anal.174, , pp.525–533, doi: 10.1016/j.jpba.2019.05.023

12. 

Li S. and Zhang B. (2013) . Traditional Chinese medicine network pharmacology: theory, methodology and application. Chin. J. Nat. Med.11, , pp.110–120, doi: 10.1016/S1875-5364(13)60037-0

13. 

Yuan H., Ma Q., Cui H., Liu G., Zhao X., Li W.et al. (2017) . How Can Synergism of Traditional Medicines Benefit from Network Pharmacology?Molecules22, , pp.E1135, doi: 10.3390/molecules22071135

14. 

Zhu J.B., Xu S., Li J., Song J., Luo B., Song Y.P.et al. (2018) . Farnesoid X receptor agonist obeticholic acid inhibits renal inflammation and oxidative stress during lipopolysaccharide-induced acute kidney injury. Eur. J. Pharmacol.838, , pp.60–68, doi: 10.1016/j.ejphar.2018.09.009

15. 

Cheung B.B., Bell J., Raif A., Bohlken A., Yan J., Roediger B.et al. (2006) . The estrogen-responsive B box protein is a novel regulator of the retinoid signal. J. Biol. Chem.281, , pp.18246–18256, doi: 10.1074/jbc.M600879200

16. 

da Silva C.A., Araujo L.S., Dos Reis Monteiro M.L.G., de Morais Pereira L.H., da Silva M.V., Castellano L.R.C.et al. (2019) . Evaluation of the Diagnostic Potential of uPAR as a Biomarker in Renal Biopsies of Patients with FSGS. Dis. Markers2019, , pp.1070495, doi: 10.1155/2019/1070495

17. 

Niculovic K.M., Blume L., Wedekind H., Kats E., Albers I., Groos S.et al. (2019) . Podocyte-Specific Sialylation-Deficient Mice Serve as a Model for Human FSGS. J. Am. Soc. Nephrol.30, , pp.1021–1035, doi: 10.1681/ASN.2018090951

18. 

Lan H., Chen W., He G. and Yang S. (2015) . miR-140-5p inhibits ovarian cancer growth partially by repression of PDGFRA. Biomed. Pharmacother.75, , pp.117–122, doi: 10.1016/j.biopha.2015.07.035

19. 

Klouche K., Amigues L., Morena M., Brunot V., Dupuy A.M., Jaussent A.et al. (2017) . On-line hemodiafiltration did not induce an overproduction of oxidative stress and inflammatory cytokines in intensive care unit-acute kidney injury. BMC Nephrol.18, , pp.371, doi: 10.1186/s12882-017-0785-1

20. 

Mainou-Fowler T., Proctor S.J. and Taylor P.R. (2004) . Interleukin 4 production by peripheral blood lymphocytes in patients with classical Hodgkin lymphoma. Leuk. Res.28, , pp.159–166, doi: 10.1016/S0145-2126(03)00216-9

21. 

Motedayyen H., Fathi F., Fasihi-Ramandi M. and Ali Taheri R. (2018) . The effect of lipopolysaccharide on anti-inflammatory and pro-inflammatory cytokines production of human amniotic epithelial cells. Reprod. Biol.18, , pp.404–409, doi: 10.1016/j.repbio.2018.09.005

22. 

Lu X.M., Ma L., Jin Y.N. and Yu Y.Q. (2015) . Lumican overexpression exacerbates lipopolysaccharide-induced renal injury in mice. Mole. Med. Rep.12, , pp.4089–4094, doi: 10.3892/mmr.2015.3940

23. 

Zhan H., Jin J., Liang S., Zhao L., Gong J. and He Q. (2019) . Tripterygium glycoside protects diabetic kidney disease mouse serum-induced podocyte injury by upregulating autophagy and downregulating beta-arrestin-1. Histol. Histopathol., pp.18097

24. 

Yu S.M., Nissaisorakarn P., Husain I. and Jim B. (2018) . Proteinuric Kidney Diseases: A Podocyte's Slit Diaphragm and Cytoskeleton Approach. Front. Med.5, , pp.221, doi: 10.3389/fmed.2018.00221

https://www.researchpad.co/tools/openurl?pubtype=article&doi=10.1042/BSR20192920&title=Anti-apoptosis mechanism of triptolide based on network pharmacology in focal segmental glomerulosclerosis rats&author=Yayu Li,Xue Jiang,Litao Song,Mengdie Yang,Jing Pan,&keyword=apoptosis,FSGS,IL4,network pharmacology,triptolide,&subject=Cell Death & Injury,Gastrointestinal, Renal & Hepatic Systems,Molecular Interactions,Research Articles,