Identification and verification of key genes related to oxidative stress in type 2 diabetes and screening of candidate drugs from Traditional Chinese Medicine

doi:10.19852/j.cnki.jtcm.2026.02.008

Abstract

Abstract:

OBJECTIVE: To identify oxidative stress (OS)-related genes involved in type 2 diabetes mellitus (T2DM) and screen potential Traditional Chinese Medicine (TCM) candidates for therapeutic use.

METHODS: Gene expression data from the GSE23343 dataset were obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between healthy and T2DM patients were identified. Weighted gene co-expression network analysis (WGCNA) was performed to select modules highly correlated with clinical traits, and core genes within these modules were identified. OS-related genes were retrieved from the GeneCards database, and the overlapping DEGs, WGCNA genes, and OS-related genes were considered as hub genes in OS-related T2DM. These hub genes were validated in GSE15653 dataset. Potential TCMs were identified by mapping the hub genes to the Coremine Medical database. In vivovalidation was performed using a T2DM rat model established by a high-fat diet and streptozotocin injection. The effects of Wedelolactone were evaluated by assessing gene expression viareal-time quantitative reverse transcription PCR (qRT-PCR) and Western blot, alongside metabolic and liver function parameters, including fasting blood glucose, glycated serum protein, insulin resistance, and lipid profiles.

RESULTS: A total of 394 DEGs (136 up-regulated and 258 down-regulated DEGs.) were identified in the GSE23343 cohort. WGCNA results showed that the turquoise module (cor = -0.56, P = 0.02) and the brown module (cor = 0.66, P = 0.004) were the most correlated with T2DM. Six hub genes [interleukin 33 (IL33), S100 calcium binding protein A8 (S100A8), Golgi membrane protein 1 (GOLM1), small nuclear ribonucleoprotein U1 subunit 70 (SNRNP70), hepatocyte growth factor activator (HGFAC), and oxidative stress induced growth inhibitor 1 (OSGIN1)] were identified, with IL33, S100A8, and GOLM1 being up-regulated, and SNRNP70, HGFAC, and GOLM1 being down-regulated. These genes distinguished T2DM from healthy controls with AUC values greater than 0.8. Experimental verification using a T2DM rat model confirmed the expression patterns of the hub genes. In vivo data demonstrated that Wedelolactone significantly reduced oxidative stress, inflammation hepatic lipid accumulation, and improved metabolic parameters such as fasting blood glucose, glycated serum protein, and insulin resistance.

CONCLUSION: These findings highlight the critical roles of IL33, S100A8, GOLM1, SNRNP70, HGFAC, and OSGIN1 as biomarkers in OS-related T2DM and suggest that Wedelolactone may be a promising TCM-based therapeutic candidate for T2DM.

Key words: diabetes mellitus, type 2, oxidative stress, gene expression profiling, computational biology, medicine, Chinese traditional, Wedelolactone

HU Jingnan, LIAO Man, XI Zhongwen, SONG Jing, WANG Yining, HE Tao. Identification and verification of key genes related to oxidative stress in type 2 diabetes and screening of candidate drugs from Traditional Chinese Medicine[J]. Journal of Traditional Chinese Medicine, 2026, 46(2): 350-359.

Figures/Tables 8

Figure 1 GO and KEGG enrichment analyses of genes in the turquoise and brown modules A: GO enrichment analysis of turquoise module genes; B: KEGG pathway enrichment analysis of turquoise module genes; C: GO enrichment analysis of brown module genes; D: KEGG pathway enrichment analysis of brown module genes. GO: gene ontology; KEGG: kyoto encyclopedia of genes and genomes.

Figure 2 Identification, expression, and diagnostic evaluation of T2DM-related hub genes. A: venn diagram showing the intersection of DEGs, WGCNA module genes, and OS-related genes; B: boxplots illustrating the expression levels of the six hub genes in liver tissues from healthy individuals (ND) and T2DM patients. The differences between groups were compared through the Wilcoxon test; C: ROC curves evaluating the diagnostic performance of the six hub genes; C1: ROC curve of GOLM1; C2: ROC curve of HGFAC; C3: ROC curve of IL33; C4: ROC curve of OSG1N1; C5: ROC curve of S100AB; C6: ROC curve of SNRNP70. DEGs: differentially expressed genes; OSGs: oxidative stress genes; ND: non-diabetes; T2DM: type 2 diabetes mellitus; ROC: receiver operating characteristic. aP < 0.01, bP < 0.05, compared to the ND group.

Table 1 Relative mRNA expression levels of six key genes in the liver tissues of control and T2DM rats ($ \bar{x} \pm s$)

Item	Control group (n = 10)	T2DM group (n = 10)
IL33	1.01±0.17	2.02±0.10^a
S100A8	1.01±0.16	2.50±0.09^a
GOLM1	1.01±0.13	1.48±0.07^a
SNRNP70	1.01±0.14	0.2 3± 0.03^a
HGFAC	1.01±0.13	0.35±0.05^a
OSGIN1	1.01±0.16	0.94±0.08^a

Table 2 Effect of Wedelolactone on glucose metabolism and insulin resistance in T2DM rats ($ \bar{x} \pm s$)

Item	Control group (n = 10)	T2DM group (n = 10)	T2DM+WED group (n = 10)
FBG	106.770±3.562	162.590±7.020^a	125.370±4.364^b
FSI	0.238±0.037	1.423±0.115^a	0.540±0.080^b
GSP	2.502±0.204	8.640±0.210^a	4.045±0.183^b
HOMA-IR	0.063±0.011	0.570±0.039^a	0.167±0.023^b

Table 3 Effect of Wedelolactone on liver function parameters in T2DM rats ($ \bar{x} \pm s$)

Item	Control group (n = 10)	T2DM group (n = 10)	T2DM+WED group (n = 10)
AST	29.570±8.080	130.650±13.689^a	68.879±5.036^b
ALT	58.617±6.476	185.643±13.449^a	96.032±6.992^b
ALP	0.106±0.007	0.207±0.007^a	0.140±0.006^b
LDH	167.580±32.556	367.489±26.839^a	172.511±15.211^b

Table 4 Effects of Wedelolactone on serum lipid profiles in T2DM rats ($ \bar{x} \pm s$)

Item	Control group (n = 10)	T2DM group (n = 10)	T2DM + WED group (n = 10)
TG	1.19±0.14	2.76±0.18^a	1.85±0.11^b
TC	542.66±80.28	1705.43±131.27^a	986.21±88.39^b
LDL-C	28.96±0.96	45.74±0.81^a	33.17±0.53^b
HDL-C	1.17±0.08	0.52±0.07^a	0.96±0.06^b

Table 5 Effect of Wedelolactone on oxidative stress markers in T2DM rats ($ \bar{x} \pm s$)

Item	Control group (n = 10)	T2DM group (n = 10)	T2DM+WED group (n = 10)
GSH	8.88±0.56	4.94±0.35^a	7.35±0.47^b
T-SOD	1.13±0.20	0.36±0.10^a	0.78±0.15^b
MDA	2.99±0.48	5.41±1.25^a	2.47±0.50^b

Figure 3 Wedelolactone modulates S100A8 expression in liver tissues of T2DM rats A: relative mRNA expression of S100A8; B: representative protein band of S100A8; C: quantification of S100A8 protein expression. Control group: rats were fed a standard diet and received daily oral gavage of physiological saline from weeks 8 to 12; T2DM group: modeled rats continued on the HFD and received daily oral gavage of physiological saline during weeks 8-12; T2DM + WED group: modeled rats received daily oral gavage of Wedelolactone (50 mg/kg) from weeks 8 to 12, while remaining on the HFD. T2DM: type 2 diabetes mellitus; HFD: high-fat diet; WED: Wedelolactone. Differences in means between groups were examined using one-way analysis of variance. Data were present as mean ± standard deviation (n = 10). aP < 0.01 compared to the Control group; bP < 0.01 compared to the T2DM group.

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