| Popis: |
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 mannitol, 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,P0.05). Conclusion Loss of Sirt1 in high glucose-treated podocytes promotes exosome secretion and podocyte injury. |
