Effects Of Jiangyatongluo Recipe On Angiotensin ?-induced Apoptosis Of Rat Renal Tubular Epithelial Cells

ObjectiveThis study is based on correlation between hypertensive nephrosclerosis and renal local activation of RAS,correlation between renal interstitial fibrosis and renal tubular epithelial cells apoptosis.Study the mechanism of renal function protection of JiangyaTongluo Formula in the treatment of hypertensive nephrosclerosis through the observation of its effect on rat renal tubular epithelial cells(NRK-52E)apoptosis induced by Angiotensin ?.Providing theoretical basis for clinical prevention and treatment of hypertensive nephrosclerosis.Methods42 SD rats were randomly divided into 3 groups:Blank Group(n=18),Valsartan Group(n=12),JiangyaTongluo Formula Group(n=12),administered by gavage for 7 days,blood was drawn from abdominal aorta and medicated serum was isolated for preparation.Rat renal tubular epithelial cells(NRK-52E)were cultured in vitro then intervened with Angiotensin ?in different concentration as 0(Control Group),10-9mol/L,10-8mol/L,10-7mol/L,10-6mol/L,10-5mol/L for 24h.Flow Cytometry instrument was implicated to detect apoptosis rate.Intervened NRK-52E with Angiotensin ? with the concentration of 10-6mol/L for 24h to induce apoptosis,then cells were cultured in different medicated medium:the Normal Control Group and the Model Group DMEM containing 10%blank rat serum;the Valsartan Group DMEM containing 10%Valsartan medicated serum;the JiangyaTongluo Formula Group DMEM containing 10%JiangyaTongluo medicated serum;the Adrenomedullin Group DMEM containing 10%blank rat serum and Adrenomedullin(10-5mmol/L);the Adrenomedullin antagonist Group DMEM containing 10%blank rat serum and Adrenomedullin antagonist(10-5mmol/L).Cell viability was determined by Cell Counting Kit-8(CCK-8)after 24h,48h and 72h.Take the cell viability value of the Normal Control Group as 100%,the relative cell viability value of each experimental group was obtained.Results1.The apoptosis rate of each concentration group was significantly higher than that of the Control Group(P<0.05,P<0.01),and the apoptosis rate increased with the increase of the concentration of Angiotensin ?.2.Compared with the Normal Control Group,the relative cell viability of each group was decreased,which of the Model Group was the lowest.Relative cell viability values after medicated medium intervention for 24h:the Valsartan Group and the Adrenomedullin Group were significantly higher than those of the Model Group and the Adrenomedullin antagonist Group(P<0.05,P<0.01);the JiangyaTongluo Formula Group was higher than those of the Model Group and the Adrenomedullin antagonist Group but the differences were not significant(P>0.05);the Adrenomedullin Group was significantly higher than that of the JiangyaTongluo Formula Group(P<0.05)but had no significant difference when compared with the Valsartan Group(P>0.05);the Valsartan Group was higher than that of the JiangyaTongluo Formula Group but the difference was not significant(P>0.05).Relative cell viability values after medicated medium intervention for 48h and 72h:the Valsartan Group,the JiangyaTongluo Formula Group and the Adrenomedullin Group were significantly higher than those of the Model Group and the Adrenomedullin antagonist Group(P<0.01);the Valsartan Group and the JiangyaTongluo Formula Group were lower than those of the Adrenomedullin Group but there were no significant differences between the three groups(P>0.05).The relative cell viability value of the Adrenomedullin antagonist Group were lower than those of the Model Group after 24h and 48h,and slightly higher than that of the Model Group after 72h,but the differences were not significant(P>0.05).Conclusion1.Angiotensin ? can induce the apoptosis of rat renal tubular epithelial cells in a concentration dependent manner.2.JiangyaTongluo Formula is able to suppress rat renal tubular epithelial cells apoptosis induced by Angiotensin ? and its effect is basically the equivalent of the positive control drug Valsartan and Adrenomedullin.3.JiangyaTongluo Formula may have the effect of reducing the apoptosis of renal tubular epithelial cells caused by renal local activation of RAS,delaying the process of renal fibrosis,and thus play a role in the clinical treatment of hypertensive nephrosclerosis.