Effect of Sirt1 on high glucose-induced exosome release from podocytes
Objective To investigate the effect of nicotinamide adenine dinucleotide-dependent deacetylase 1(Sirt1)on high glucose-induced exosome release from podocytes.Methods Immortalized mouse podocytes MPC5 were divided into six groups:normal glucose group(5.5 mmol/L glucose,group A),high mannitol group(5.5 mmol/L glucose+24.5 mmol/L manni-tol,group B),high glucose group(30.0 mmol/L glucose,group C),high glucose+Sirt1-overexpressed lentivirus transfection group(Sirt1-overexpressed lentivirus transfection+30.0 mmol/L glucose,group D),high glucose+negative lentivirus transfection group(negative lentivirus transfection+30.0 mmol/L glucose,group E),and high glucose+exosome secretion inhibitor group(GW4869+30.0 mmol/L glucose,group F).Western blot was used to analyze the expression levels of Nephrin,Podocin,Sirt1,CD63,CD81,and Alix in each group.Real-time quantitative polymerase chain reaction was used to analyze the expression level of Sirt1 mRNA in D and E group.The morphology of podocyte exosomes was observed by a transmission electron microscope.The particle size and concentration of exosomes were determined by nanoparticle tracking analysis.Results The results of RT-qPCR showed that the relative expression of Sirt1 mRNA was significantly increased in group D compared with that in group E(t= 14.580,P＜0.01).The results of nanoparticle tracking analysis and Western blot showed that the relative expression of Sirt1,Nephrin,and Podocin proteins in podocytes among groups A to C was significantly different(F=49.84-106.40,P＜0.01).Com-pared with group A,group C had significantly increased secretion of podocyte exosomes(t=14.550,P＜0.01)and significantly re-duced expression of Sirt1,Nephrin,and Podocin(t=7.446-15.110,P＜0.01).Compared with group E,group D had significantly reduced release of podocyte exosomes(t=74.610,P＜0.01)and significantly increased relative expression of Sirt1,Nephrin,and Podocin(t=4.657-32.860,P＜0.05).Compared with group C,group F had significantly reduced release of podocyte exosomes(t=16.300,P＜0.05)and significantly increased relative expression of Nephrin and Podocin(t=3.790,8.151,P＜0.01),but showed no significant change in the expression level of Sirt1(P＞0.05).Con-clusion Loss of Sirt1 in high glucose-treated podocytes promotes exosome secretion and podocyte injury.
国家科技期刊平台
NSTL
万方数据
维普
