Protective Effect Of Astragaloside ? On Human Mesangial Cells And Its Mechanism Under High Glucose Conditions

Objectives To observe the damage of human glomerular mesangial cells and preventive and therapeutic effects of Astragaloside ? under high glucose conditions in vitro.To study the protective effect of astragaloside ? on human glomerular mesangial cells injury and its possible mechanism,observe the regulatory effect of Nrf2 signaling pathway.To explore the possible pathogenesis of diabetic nephropathy,to provide a new theory,a new target and a theoretical basis for new drugs.Methods1.HMCs were randomly divided into the following two groups:(1)normal contorl(NG)group,(2)high glucose(HG)group,the proliferation of each group was detected at 12 h,24 h,36 h,48 h and 72 h by MTT assay.2.HMCs were randomly divided into the following six groups as the time group:(1)NG group,(2)HG 6 h group,(3)HG 12 h group,(4)HG 24 h group(5)HG 36 h group,(6)HG 48 h group.The m RNA expression and protein levels of Nrf2,HO-1,i NOS,ICAM-1 were determined by q PCR and Western Blotting,respectively.The supernatant were collected to measure the amount of H2O2,MDA and the activity of T-SOD and GSH-Px.3.HMCs were randomly divided into the following six groups:(1)NG group,(2)HG group,(3)HG+ AS-? 10 ?mol/L group,(4)HG+ AS-? 25 ?mol/L group,(5)HG+ AS-? 50 ?mol/L group,(6)HG+ AS-? 100 ?mol/L group,all groups were treated for 48 h.The effect of AS-? in HMCs proliferation were detected by MTT assay,the m RNA expression and protein levels of Nrf2,HO-1,i NOS,ICAM-1 were determined by q PCR and Western Blotting,respectively.The supernatant were collected to measure the amount of H2O2,MDA and the activity of T-SOD and GSH-Px.Results1.Compared with normal group,MTT results showed that HMCs were significantly proliferation in high glucose conditions.Compared with high glucose group(HG),AS-? significantly inhibited the proliferation of HMCs in dose-dependent.2.Western blot and q PCR results showed that Nrf2 m RNA and protein expression levels were increased at different time points,HO-1 m RNA and protein expression levels increased at 12 h and 24 h,then at 36 h and 48 h decreased to normal expression level compared with control group.Compared with high glucose group,AS-? increased Nrf2 and HO-1 expression and suppressed the overexpression of i NOS and ICAM-1 in HMCs under high glucose conditions.3.The results showed that the amount of H2O2 and MDA were increased and the activity of T-SOD and GSH-Px were reduced in supernatant at different time points.Compared with HG group,after 48 h of AS-IV treatment,the above phenomenon was completely reversed.Conclusion AS-?could significantly inhibit HMCs excessive proliferation in high glucose conditions.The mechanism maybe related to the activating of Nrf2 pathway,upregulating Nrf2,HO-1 m RNA and protein expression,downregulating m RNA and protein expression of i NOS,ICAM-1,reducing the amount of H2O2 and MDA,and increasing the activity of T-SOD and GSH-Px in supernatant.