Effect Of Huihuigansongyin On Glomerular Mesangial Cells And Endothelial Cells Induced By High Glucose And Its Autophagy Mechanism

Objective To verify the early protective effect of huihuigansongyin on glomerular mesangial cells and glomerular endothelial cells under the condition of high glucose,and to explore the molecular mechanism of huihuigansongyin to improve glomerular damage by regulating the related autophagy signal pathway(timp3-stat1-foxo1 signal pathway)in mesangial cells and the related autophagy signal pathway(TGF-? 1-bambi)in endothelial cells The pathogenesis and treatment target provide new ideas.Methods immortalized mouse glomerular mesangial cells(SV40 MES 13)and mouse glomerular endothelial cells(mgecs)were selected.30 mmol / l high glucose solution was used as the high glucose model group.The two kinds of cells were divided into 5 groups.Blank serum and huihuigansongyin containing serum were used for intervention.CCK-8 method was used to detect the SV40 MES 13 induced by different containing serum,MGECs 2.Flow cytometry was used to detect the cell cycle and intracellular ROS of SV40 MES 13 and MGECs 2 cells,MDC fluorescence staining was used to detect the autophagy rate of SV40 MES 13 and MGECs cells,Western blot was used respectively;WB)technique was used to detect the expression of TIMP3,STAT1,FoxO1,TGF-?1,Bambi,LC3 and p62 in mesangial cells and endothelial cells.Result1.The results of HGY on the function of glomerular mesangial cells in high glucose stateThe results of CCK-8 showed that HGY had a certain inhibitory effect on the proliferation of mesangial cells compared with the model group at 48 hours after intervention,and the inhibition effect of high dose group was the most obvious(P < 0.01).The cell cycle showed that the high dose HGY group could cause G1 /S arrest of mesangial cells compared with the model group(P < 0.01).Flow cytometry showed that ROS level of HGY high dose group was lower than that of model group(P < 0.01).The incidence of autophagy was observed by MDC staining: the fluorescence intensity and quantity of HGY high dose group were higher than that of mod group(P < 0.01).2.HGY effects on the function of glomerular endothelial cells in high glucose conditionAt 48 hours after the intervention of HGY group,the OD value of high dose group was significantly lower than that of model group,which had a certain inhibitory effect on the proliferation of glomerular endothelial cells(P < 0.01).Cell cycle showed that the proportion of cells in G1 and S phase in HGY high dose group was higher than that in model group(P < 0.01).Flow cytometry showed that ROS level of HGY high dose group was lower than that of model group(P < 0.01).The incidence of autophagy was observed by MDC staining: the fluorescence intensity and quantity of HGY high dose group were higher than that of mod group(P < 0.01).3.The results of molecular mechanism of HGY regulating autophagy in mesangial cells under high glucose conditionWestern blot showed that compared with the model group,the expression of LC3 in HGY group was significantly higher(P < 0.01),and the expression of p62 was lower(P < 0.01);compared with the model group,the expression of TIMP3,STAT1 and FoxO1 protein decreased(P < 0.05).4.Research results on the molecular mechanism of HGY regulating autophagy in endothelial cells under high glucose conditionWestern blot showed that the expression of LC3 in HGY was higher than that in model group,and the expression of p62 in HGY was higher than that in HGY and tended to blank group(P < 0.01).Compared with the model group,the expression of TGF-?1 in HGY group decreased(P < 0.05),and the expression of Bambi protein increased(P < 0.05).Conclusion1.HGY has protective effect on the early stage of mesangial cells and endothelial cells in high glucose state2.HGY can down regulate the expression of TIMP3,STAT1 and FoxO1 protein,activate the TIMP3-STAT1-FoxO1 pathway,promote the autophagy of mesangial cells and improve the process of glomerular damage.3.HGY can up regulate the expression of BAMBI down regulate TGF-?1,activate TGF-?1-BAMBI pathway,promote autophagy and reduce glomerular damage.