Effects of rapamycin on proliferation,apoptosis and IL-6 expression of human glomerular podocytes induced by high glucose
Objective To observe the effects of rapamycin on proliferation,apoptosis,and IL-6 expression of hu-man glomerular podocytes induced by high glucose,and to explore the related mechanisms based on phosphatidylinositol-3-kinase(PI3K)/serine-threonine kinase(AKT)signaling pathway.Methods The human glomerular podocyte HGPC cell line was cultured in vitro and divided into the control group,high glucose group,and experimental group,respective-ly.Cells in the control group were not intervened,while cells in the high glucose group and the experimental group were added with 30 mmol/L D-glucose,and the experimental group were added with 2.5,5,and 10 µmol/L rapamycin on the basis of the high glucose group.The cell viability of each group was detected to screen the optimal concentration of rapamy-cin.HGPC cells were divided into the control group,high glucose group,experimental group,inhibitor group,rapamycin + inhibitor group,and rapamycin + agonist group,respectively.Cells in the control group were not added with any drugs,cells in the high glucose group were added with 30 mmol/L D-glucose,cells in the inhibitor group were added with 30 mmol/L D-glucose +10 µmol/L LY294002,a PI3K/AKT signaling pathway inhibitor,cells in the experimental group,ra-pamycin + inhibitor group and rapamycin + agonist group were added with 30 mmol/L D-glucose and 10 µmol/L rapamy-cin,cells in the rapamycin + inhibitor group were added with 10 µmol/L LY294002,and cells in the rapamycin + agonist group were added with 10 µmol/L PI3K/AKT signaling pathway agonist SC79.Each group was cultured for 24 h after ad-ministration.Cell proliferation was detected by 5-ethynyl-2'-deoxyuridine assay,and apoptosis was detected by Hoechst 33258 staining.Interleukin-6(IL-6)in the supernatant of cell culture medium was detected by ELISA.Western blotting was used to detect Cyclin D1,Caspase-3 and PI3K/AKT pathway-related proteins in cells.Results The cell prolifera-tion rate and Cyclin D1 expression in the high glucose group were lower than those in the control group,and the cell prolif-eration rate and Cyclin D1 expression in the experimental group and inhibitor group were higher than those in the high glu-cose group;compared with the experimental group,the cell proliferation rate and Cyclin D1 expression increased in the ra-pamycin + inhibitor group,and decreased in the rapamycin + agonist group(all P<0.05).The apoptosis rate,Caspase-3 expression and p-PI3K/PI3K,p-AKT/AKT and IL-6 levels in the high glucose group were higher than those in the control group,and the apoptosis rate,Caspase-3 expression and p-PI3K/PI3K,p-AKT/AKT and IL-6 levels in the experimental group and inhibitor group were lower than those in the high glucose group.Compared with the experimental group,the apoptosis rate,Caspase-3 expression and the levels of p-PI3K/PI3K,p-AKT/AKT and IL-6 in the rapamycin + inhibitor group decreased,and the apoptosis rate,Caspase-3 expression and the levels of p-PI3K/PI3K,p-AKT/AKT and IL-6 in-creased in the rapamycin + agonist group(all P<0.05).Conclusion Rapamycin can promote the proliferation of human glomerular podocytes induced by high glucose,reduce apoptosis and inflammatory injury,and the mechanism may be relat-ed to the inhibition of PI3K/AKT pathway.
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