Effect of hesperidin on high glucose induced autophagy in glomerular mesangial cells by regulating the mTOR/HIF-1α pathway
[Objective]To investigate the effect of hesperidin on autophagy in glomerular mesangial cells induced by high glucose by regulating the mammalian target of rapamycin(mTOR)/hypoxia-inducible factor-1α(HIF-1α)pathway.[Methods]SV40 MES-13 cells with good growth were randomly separated into control group(without treatment),mannitol group(30 mmol/L mannitol treatment),high glucose group(30 mmol/L glucose),different doses hesperidin groups(30 mmol/L,60 mmol/L hesperidin treatment),and 60 mmol/L hesperidin+insulin-like growth factor-1(IGF-1)group(60 mmol/L hesperidin,100 ng/mL IGF-1 treatment).Methylthiazolyldiphenyl-tetrazolium bromide(MTT)and 5-ethynyl-2'-deoxyuridine(EdU)were used to detect cell proliferation.Enzyme linked immunosorbent assay(ELISA)was applied to detect levels of tumor necrosis factor-α(TNF-α)and interleukin(IL)-1 β.Flow cytometry was applied to detect the level of cell apoptosis.Real-time quantitative PCR(qRT-PCR)was applied to detect the mRNA expression levels of microtubule-associated protein 1 light chain 3(LC3),p62,and autophagy related gene 5(ATG5).Western blot was applied to detect the expression levels of p-PI3K,p-Akt,p-mTOR,PI3K,Akt,mTOR,and HIF-1α related proteins.[Results]Compared with the control group,there was no statistically obvious difference in absorbance value,EdU positivity rate,TNF-α,IL-1β levels,apoptosis rate,LC3,p62,ATG5 mRNA expression,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,and HIF-1α expression in the mannitol group(P＞0.05).After high glucose induction,the absorbance value,EdU positivity rate,TNF-α,IL-1β levels,p62 mRNA expression,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,HIF-1α expression were obviously increased compared to the control group and mannitol group,while apoptosis rate,LC3,and ATG5 mRNA expression were obviously decreased(P＜0.05).After treatment with different doses of hesperidin,the absorbance value,EdU positive rate,TNF-α,IL-1β levels,p62 mRNA expression,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,and HIF-1α expression were obviously reduced compared to the high glucose group.The apoptosis rate,LC3,and ATG5 mRNA expression were obviously increased,and there was a difference between groups(P＜0.05).After treatment with 60 mmol/L hesperidin combined with IGF-1,the absorbance value,EdU positive rate,TNF-α,IL-1β levels,p62 mRNA expression,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,and HIF-1α expression were obviously increased compared to 60 mmol/L hesperidin alone treatment.The apoptosis rate,LC3,and ATG5 mRNA expression were obviously reduced(P＜0.05).[Conclusion]Hesperidin promotes high glucose induced autophagy in mesangial cells and inhibits their proliferation by regulating the mTOR/HIF-1α pathway.
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