人参皂苷Rh1通过激活Nrf2/SLC7A11/GPX4通路减轻高糖诱导的肾小管上皮细胞损伤OA

AIM:To investigate the effect and underlying mechanism of ginsenoside Rh1(G-Rh1)against high glucose(HG)-induced injury in the human renal tubular epithelial cell line HK2 cells.METHODS:HK2 cells were exposed to 25 mmol/L glucose to establish a cellular injury model.Cells were assigned to the following groups:model(HG)group,nuclear factor erythroid 2-related factor 2(Nrf2)agonist NK252-treated(HG+NK252)group,Nrf2 inhibi-tor ML385-treated(HG+ML385)group,G-Rh1 intervention(HG+G-Rh1)group,and inhibitor+intervention(HG+ML385+G-Rh1)group.HK2 cells cultured in DMEM containing 5.5 mmol/L glucose served as the normal control(NC)group.Cell viability was assessed using the CCK-8 assay.Levels of reactive oxygen species(ROS),malondialdehyde(MDA),lactate dehydrogenase(LDH)activity,and ferrous iron(Fe2+)content were measured using commercial assay kits.The mRNA and protein expression levels of Nrf2,solute carrier family 7 member 11(SLC7A11),and glutathione peroxidase 4(GPX4)were analyzed by RT-qPCR and Western blot,respectively.RESULTS:Compared with the NC group,HK-2 cells cultured under high-glucose conditions for 48 h exhibited significantly reduced viability(P<0.01),accompanied by elevated intracellular ROS and MDA levels,increased LDH activity and Fe2+content(P<0.01),as well as downregulated mRNA and protein expression of Nrf2,SLC7A11,and GPX4(P<0.01).Relative to the HG group,the HG+G-Rh1 group showed markedly improved cell viability(P<0.01),the HG+G-Rh1 and HG+NK252 groups showed markedly reduced ROS,MDA,LDH,and Fe2+levels(P<0.01,P<0.05),and upregulated expression of Nrf2,SLC7A11,and GPX4 at both mRNA and protein levels(P<0.01).In contrast,the HG+ML385 group exhibited further in-creases in ROS,MDA,LDH,and Fe2+(P<0.01,P<0.05),decreased SLC7A11 protein expression,and increased GPX4 protein expression(P<0.01).Compared with the HG+G-Rh1 group,the HG+ML385+G-Rh1 group showed exacer-bated oxidative stress and ferroptosis-related indices,along with suppressed Nrf2,SLC7A11,and GPX4 expression at both transcriptional and translational levels(P<0.01).CONCLUSION:Ginsenoside Rh1 attenuates high glucose-in-duced oxidative damage and ferroptosis in HK-2 cells,likely through upregulation of Nrf2 and subsequent activation of the Nrf2/SLC7A11/GPX4 signaling pathway.