Study On The Effect Of Qianyang Yuyin Granules On Hypertensive Renal Damage By Influencing The Level Of Podocyte Autophagy

AIMThe purpose of this study was to investigate the role of renal podocyte autophagy in hypertensive renal damage,to study the therapeutic effect of Qianyang Yuyin Granules(QYYY G)on early renal damage of hypertension,and to confirmed the relationship between the autophagy level of renal podocytes and QYYY G through in vitro and in vivo experiments.This research result can make QYYY G be promoted better and provide new ideas for the treatment of hypertension with traditional Chinese medicine.MethodsExperiment 1:In this study,spontaneous hypertension rats(SHR)were used as the research object,and QYYY G were used for intervention.During the experiment,the blood pressure of the rats was observed,and the urine and blood were collected to detect the metabolic indicators related to renal damage.HE staining was used to understand the pathological structure of glomerulus and renal tubules in rats.The morphology,organelle changes and number of autophagosomes of glomerular podocytes were observed by transmission electron microscopy.This experiment can observe the renal damage caused by long-term hypertension and explore the improvement effect of QYYY G on the damage of glomerular podocyte cells in hypertensive rats.Experiment 2:Medicated serum of SD rats was collected by the intervention of QYYY G and valsartan.Podocytes were cultured in vitro.This experiment determined the Ang II concentration which can induce the damage of podocytes.The cells were interfered with the medicated serum of QYYY G and valsartan.Western Blot were used to investigate the autophagy level of podocytes.Experiment 3:Immunofluorescence technique was used to observe the effect of QYYY G medicated serum on TRPC 6 and p-mTOR protein.Intracellular Ca2+level was detected by calcium imaging.Western Blot were used to investigate the changes of p-AMPK/AMPK in podocytes.To understand the potential target of the medicated serum of QYYY G on podocytes autophagy,and lay a foundation for exploring the mechanism of improving hypertensive renal damage with QYYY G.ResultsExperiment 1:The blood pressure and related metabolic indexes of rats in SHR group were significantly higher than those in WKY group.The glomeruli were obviously atrophied,the renal tubules were dilated,more fibroid-like tissues were generated in the renal cortex,the autophagosomes in podocytes were increased,and the foot processes were extensively fused.The medicated serum of QYYY G can significantly reduce the blood pressure level of SHR rats,and have a better improvement effect on the pathological damage of glomerular and podocytes which caused by hypertension.The efficacy is dose-related.Experiment 2:Ang ? have obvious inhibitory effect on podocytes proliferation.It can significantly improve the protein expression of LC3 ?/LC3 ? and Beclin in podocytes,and inhibit the protein expression of P62.The medicated serum of QYYY G may protect podocytes from damage caused by Ang ?,reduce LC3 ?/LC3 ? and Beclin protein expression,increase the P62 protein expression.It can significantly improve the autophagy of podocytes induced by Ang ?.Experiment 3:Ang ? can significantly improve the protein expression of TRPC 6 and increase intracellular Ca2+concentration.At the same time,Ang ? affects the expression of autophagy related proteins in podocytes.The medicated serum of QYYY G can inhibit the TRPC 6 protein expression,reduce the concentration of intracellular Ca2+ and inhibit podocytes autophagy.Conclusion(1)QYYY G has a better improvement effect on renal pathological damage which caused by hypertension(2)Elevated autophagy of renal podocytes induced by hypertension may be involved in the formation of hypertensive renal damage(3)QYYY G can may influence the intracellular Ca2+level through TRPC 6 protein in podocytes,and then regulating autophagy...