Effects Of H2s On High Concentrated Glucose-induced Autophagy Of Glomerular Mesangial Cell And Possible Molecular Mechanisms

Objective: To observe effects of H₂S on high concentrated glucose-induced autophagy of glomerular mesangial cell and its possible molecular mechanisms.Methods: Glomerular mesangial cells from mice were treated by sodium hydrosulfide?Na HS,a donor of H₂S?with various concentrations for 48 h.Using MTT test and Brdu test respectively detected the cell viability and cell proliferation to determine given H₂ S with suitable concentration.Experimental grouping was as follow:group of control?conventional culture medium?,group of high concentrated glucose solution,group of H₂S?conventional culture medium?,group of high concentrated glucose solution and H₂ S,group of high concentrated glucose solution and Rapamycin?autophagy accelerant?,group of high concentrated glucose solution,Rapamycin and H₂ S.Expressive levels of associated protein related to autophagy,such as Microtubule-associated protein 1 light chain 3?MAP1-LC3,LC3?,Beclin-1,Sequestosome-1?SQSTM1/p62?,and phosphorylated protein such as AMPK and m TOR in each group were examined with Western blot.Autophagosome change in each group were also determined with immunofluorescence and transmission electron microscope.Changes of reactive oxygen species?ROS?level were investigated with ROS kit.Results: Brdu and MTT results showed that given H₂ S with 300 ?M concentration in experiment did not influence on cell viability or on cell proliferation,which was the optimal concentration.Western blot told that after high concentratedglucose treated cell,expressive levels of LC3 and Beclin-1 were increased but those of P62 were decreased compared with control group.Cells pretreatment with H₂ S could remarkably diminish autophagy high levels induced by high concentrated glucose.Cells pretreatment with Rapamycin could obviously inhibit effect of H₂ S on autophagy high levels induced by high concentrated glucose.Results from immunofluorescence test and transmission electron microscope were correspondent to those of Western blot.Moreover,phosphorylated levels of autophagy pathway protein named AMPK did not significantly change in each group but those of another protein named m TOR showed clearly increase after cell treated by H₂ S.In addition,expressive levels of ROS in group of high concentrated glucose solution were significantly increased compared with group of control and cell treated by H₂ S could greatly decline expressive levels of ROS in group of high concentrated glucose solution and H₂ S and cell treated by Rapamycin could significantly elevate ROS expressive levels in group of high concentrated glucose solution,Rapamycin and H₂ S.Conclusion: High concentrated glucose could result in high expressive levels of autophagy and ROS in glomerular mesangial cell.H₂ S could reduce high responses of autophagy and ROS induced by high concentrated glucose and its mechanisms may be involved in activating m TOR signaling,lowering ROS levels and then displaying effects in glomerular mesangial cell.