Experimental Study On The Effect Of Jiangya Tongluo Prescription On Regulating PAkt Expression And Inhibiting NRK-52E Apoptosis Induced By Ischemia And Hypoxi

ObjectiveIn this study,we established the apoptosis of renal tubular epithelial cells(NRK-52E)model to imitate the pathological process of hypertensive renal injury which was caused by hypoxia-ischemia.And observed the effect of JiangYaTongLuo Formula on the apoptosis of NRK-52E from molecular biology perspective,investigated the acting mechanism of JiangYaTongLuo Formula on the treatment of hypertensive renal damage,to provide experimental basis for the clinical application of JiangYaTongLuo Formula.Methods42 SD rats were randomly divided into 3 groups:Blank Control Group(n=18),Traditional Chinese Medicine Group(n=14),and Western Medicine Group(n=10).They were respectively administered by gavage the same volume of normal saline,JiangYaTongLuo Formula,and valsartan for seven days,then took blood from abdominal aorta for the preparation ofdrug-containing serum.(1)Normally cultured NRK-52E was modeled for ischemia and hypoxia,and detected the cell viability values of the normal and model groups respectively with CCK8 kit after 8h,10h,and 12h,finally got 12h as the best modeling time.(2)The cells were randomly divided into 4groups:normal group,model group,valsartan group,and JiangYaTongLuo Formula group.Then established the apoptosis of NRK-52E after 12h of hypoxia-ischemia,added corresponding drug-containing serum to intervene the cells,then detected the cell viability values with CCK8 after 18h,24h,and 48h,finally chose 24h as the best intervention time.(3)The cells were randomly divided into the above four groups,intervened them with the best model time and dosing time.Then detected the expression of PAkt in the 4 groups by immunohistochemistry.Results(1)Compared with the normal group,the cell viability of the model group at 8h,10h,12h after ischemia and hypoxia were decreased significantly(both P<0.01),calculated cell survival rate based on the cell viability value,found the cell survival rate decreased with the prolongation of the modeling time.(2)Intervened NRK-52E with drug-containing serum,the cell viability values of the normal group after 18h,24h,and 48h of dosing were significantly higher than those of the other three groups(P<0.01).After 18h of dosing,the cell viability values of valsartan group(P<0.05)and JiangYaTongLuo Formula group(P>0.05)were all lower than those of the model group.After 24h of dosing,the cell viability values of the valsartan group and the JiangYaTongLuo Formula group were both significantly higher than the model group(both P<0.05),but there was no significant difference between the valsartan group and the JiangYaTongLuo Formula group(P>0.05);After 48h of dosing,the cell viability values of the valsartan group and JiangYaTongLuo Formula group were both higher than that of the model group,but neither of them had significant difference(both P>0.05).(3)Immunohistochemistry:PAkt was highly expressed in the normal group and low in the model group.There was a significant difference between the normal group and each group(P<0.05).The expression of PAkt in the valsartan group was lower than that in the model group,but there was no significant difference between the two groups(P>0.05).The expression of PAkt was significantly higher in the JiangYaTongLuo Formula group than in the model group(P<0.01).Conclusion(1)Ischemia and hypoxia can promote the apoptosis of NRK-52E to establish hypertensive renal damage model,which in a time-dependent manner.(2)JiangYaTongLuo Formula can inhibit the apoptosis of NRK-52E which induced by ischemia and hypoxia,its mechanism of action is achieved by up-regulating the expression of PAkt,which is of great significance in delaying the progression of hypertensive renal damage.The study provide experimental evidence for the clinical ues of JiangYaTongLuo Formula treatment of hypertensive renal damage.