丹瓜护脉口服液通过HIF-1α调控铜死亡保护2型糖尿病血管内皮细胞的机制研究OA

Objective To investigate the mechanism of Dangua Humai Oral Liquid(DGHM)in improving copper death(cuproptosis)of vascular endothelial cells in type 2 diabetes mellitus(T2DM)rats through hypoxia-inducible factor-1α(HIF-1α).Methods Network pharmacology was used to screen the active components,potential targets of DGHM,and its intersection targets with T2DM and oxidative stress.Protein-protein interaction network construction and GO and KEGG enrichment analyses were performed.A T2DM rat model was induced by a high-fat and high-sucrose diet combined with streptozotocin injection.The rats were randomly divided into normal group,model group,DGHM group,HIF-1α inhibitor(PX-478)group,DGHM combined with PX-478 group,and Metformin control group.After 14 weeks of intervention,glycolipid metabolism indicators including FBG,2 hBG,HbA1c,TC,TG,HDL-C and LDL-C were assessed by biochemical methods and ELISA;the morphological structure of the thoracic aorta was observed by HE staining;HIF-1α protein expression in thoracic aortic tissue was detected by immunohistochemistry;vascular endothelial function and oxidative stress markers including endothelin-1(ET-1),vascular endothelial growth factor(VEGF),Caspase-3 and nitric oxide synthase(NOS)were measured by ELISA;copper deposition was detected by rubeanic acid staining.Results Network pharmacology identified 224 intersection targets with T2DM and oxidative stress,with HIF-1α being one of the core targets.Animal experiments showed that DGHM could significantly reduce FBG,2 hBG,HbA1c,TC,TG,LDL-C,ET-1 and Caspase-3 levels,increase HDL-C,VEGF,NOS activity,and HIF-1α protein expression of model rats(P<0.05,P<0.01,P<0.001),and alleviate structural damage and copper deposition in the thoracic aortic endothelium.After inhibition of HIF-1α,glucose and lipid metabolism disorders,oxidative stress,abnormal copper ion aggregation,and vascular endothelial damage worsened.The combination of DGHM and PX-478 could improve to some extent the glucose and lipid metabolism disorders,oxidative stress,and copper overload caused by PX-478.Conclusion DGHM may exert a protective effect on T2DM vascular endothelium by upregulating HIF-1α expression,reducing abnormal accumulation of copper ions,inhibiting copper death process,alleviating oxidative stress caused by it.