芪蒡糖肾方调控Nrf2/Gpx4通路改善糖尿病肾病肾纤维化的机制研究OA

Objective:To investigate the protective effect of Qibang Tangshen Formula(QBTSF)on diabetic nephropathy mice and high glucose-induced fibrosis of human renal cortical proximal tubular epithelial cells HK2.Methods:(1)In vivo experi-ments:C57BL/6 J male mice were selected to establish an animal model of diabetic nephropathy by using high-sugar and high-fat feed+streptozocin injection,and after successful modeling,the mice were divided into a normal group,a model group,a dapa-gliflozin group,a QBTSF low-dose group,a QBTSF middle-dose group,and a QBTSF high-dose group.The drug was adminis-tered continuously for 12 weeks,and the levels of 24 h urinary protein(24 hpro),serum creatinine(Scr),fasting blood glucose(FBG),and ferrous ion(Fe2+)in renal tissues were measured in each group.Masson staining was used to observe the degree of renal histopathological damage.Western blot was used to detect the expression levels of Nrf2/Gpx4 signaling pathway and down-stream fibrosis-related proteins.(2)In vitro experiments:CCK-8 was used to detect cell viability and screen the optimal interven-tion concentration of QBTSF.Western blot was used to detect the expression level of Col3a1 and determine the modeling condi-tions.The HK2 cells were divided into a normal group,a high glucose group,a QBTSF group,and a dapagliflozin group.The in-tracellular concentrations of malondialdehyde(MDA),glutathione(GSH),Fe2+,and reactive oxygen species(ROS)were mea-sured,and the expression levels of Nrf2/Gpx4 signaling pathway and fibrosis-related proteins were analyzed.Transmission elec-tron microscopy was used to observe the ultrastructural changes of HK2 cells.Results:(1)In vivo experiment:Compared to the model group,after the intervention of QBTSF,24 hpro,FBG,Scr,and Fe2+levels in renal tissues were decreased(P＜0.01).In addition,QBTSF could promote the expression of Nrf2,Gpx4,E-Cadherin,and Slc7a11 proteins,as well as reduce the ex-pression of α-SMA、Col3a1,and Vimentin proteins in renal tissues(P＜0.05).(2)In vitro experiments:Compared to the high glucose group,QBTSF could significantly reduce intracellular MDA,Fe2+,and ROS concentrations and improve mitochondrial damage while increasing intracellular GSH content(P＜0.01).In addition,QBTSF promoted the expression of intracellular Nrf2,Gpx4,Slc7a11,and E-Cadherin proteins,while inhibiting the expression of α-SMA,Col3a1,and Vimentin proteins(P＜0.01).Conclusion:QBTSF helps to improve renal fibrosis in diabetic nephropathy,which may be associated with the activation of Nrf2/Gpx4 signaling pathway.