Effect of astragaloside Ⅳ on renal cell damage induced by deoxysphinine
Objective To establish a model of renal cell injury induced by deoxysphinine(doxSA)and explore the effect of astragaloside Ⅳ(AST)on doxSA induced renal cell injury by regulating the nuclear factor erythroid-2 related factor 2(Nrf2)pathway.Methods HK-2 cells were cultured and their activity was detected using cell counting kit-8(CCK-8).They were randomly divided into control group(Cont),doxSA group,doxSA+AST group,and AST group based on whether doxSA and AST were added.After intervention,CellTiter-Glo® 2.0 was used to measure ATP content in kidney epithelial cells;immunofluorescence was used to detect mitochondrial activity and morphology;polymerase chain reaction(PCR)was performed to detect the mRNA levels of heme oxygenase 1(HO-1)and NAD(P)H dehydrogenase quinone 1(NQO1);and oxidative stress was measured in kidney epithelial cells by transfection.Analysis of variance(ANOVA)was used to assess the statistical significance of the differences between the groups.Results At a concentration of 62.5 μg ·mL-1,doxSA significantly injured kidney cells,and the subsequent experiments used 62.5 μg·mL-1 of doxSA to treat HK-2 cells to establish the doxSA model.The ATP content(percentage)of the Cont,doxSA,doxSA+AST,and AST groups was(100.00±2.21)％,(44.06±3.61)％,(64.51±4.20)％,and(100.00±14.21)％respectively.There was a statistically significant difference in ATP content among the four groups(F=38.718,P＜0.001).The detection of mitochondrial function and morphology showed that the toxic effect of doxSA on cells was mainly due to mitochondrial damage,characterized by decreased mitochondrial activity and reduced number.AST intervention could restore the activi-ty and morphology of the damaged mitochondria and protect them.The results of the measurement of oxidative stress in cells showed that compared to the Cont group,the heat value was higher in the doxSA group,indicating significant oxida-tive stress;compared to the doxSA group,the heat value decreased in the doxSA+AST group,suggesting that AST in-tervention significantly reduced the level of cellular oxidative stress.The expression levels of Nrf2 downstream genes HO-1 and NQO1 mRNA in kidney epithelial cells of the doxSA group were 0.47±0.03 and 0.34±0.01,which were reduced by more than 50％compared to the normal Cont group's 0.98±0.07 and 0.96±0.12(P＜0.001).After AST interven-tion,the expression values of HO-1 and NQO1 mRNA in cells were 1.37±0.16 and 1.72±0.12,which increased com-pared to the doxSA group's 0.47±0.03 and 0.34±0.01,with a statistically significant difference(P=0.019),indicating that the AST intervention activated the Nrf2 pathway in cells.Conclusions AST significantly increases the expression and mitochondrial activity of HO-1 and NQO1,exerting antioxidant effects.AST activates the antioxidant pathway mediated by the Nrf2 pathway,exerting a renal protective effect.
astragaloside Ⅳkidneyoxidative stressnuclear factor erythroid-2 related factor 2
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