Research Of Mechanism Of TRPC6 Podocytes Under Acute Hypertension Condition

Background:There are three innate cells in glomerulus:mesangial cells,endothelial cells,podocytes.Above all,podocytes are terminally differentiated cells,which have steady phenotype.Podocytes injures is a significant link of chronic renal disease.The internal flow of Ca2+ activate downstream signaling pathway is an activate factor of podocytes injures.TRPC6 is an important channel which regulate the internal flow of Ca2+.The TRPC6 channel functions activate in the disease state and therefore Ca2+internal flow.Mitochondria are the main energy processing plants in the body,providing more than 90 percent of the energy of the body,producing a large amount of oxygen free radicals,inducing apoptosis,and related to various cardiovascular diseases.At present,we suspect that in the induction of acute hypertension,TRPC6 regulates intracellular Ca2+,which leads to mitochondrial Ca2+ overload,which leads to mitochondrial disintegration and apoptosis.We proposed to establish acute hypertension podocyte damage model in mice,using genetic interference technology,specific protein/channel inhibitors,understand TRPC6 regulate Ca2 + sertoli cell damage caused by internal flow,the mechanism of action;The network regulation mechanism of TRPC6 was clarified through the detection of Ca2+-dependent signal pathway.In vitro and in vivo intervention,exploring target blocking TRPC6 channel to reduce the possibility of podocytes injury may provide a new theoretical basis for clinical treatment.Objectives:This research adopts the "In vivo cryotechnique" to deal with the mice kidney tissue samples.It aims to keep the TRPC6 of the mice glomerular podocyte in situ by immunohistochemistry and immunofluorescence,Real Time PCR and likely technology in abnormal hemodynamics,especially in cases of acute hypertension.It helps to discover dynamic change of the distribution of the TRPC6 on the podocytes.Methods:1.Experimental animal:10 Kunming mouse,25-35 g weight,male,bought from charles river.2.Experimental grouping:(1)Normal hemodynamic status group under IVCT condition(2)Acute hypertension group under IVCT condition(Ligate the abdominal aorta under the left renal artery of the mouse in 15 minutes.)Results:1.The structure of glomerular under various hemodynamic conditions:the capilla-ry loops are smooth and plump in normaltensive condition,however,in the acute hypertensive condition,the Bowman?s space and luminal spaces of the proximal or the distal tubules became more widely opened in the renal cortices.2.The distribution changes of TRPC6 in glomerulars:TRPC6 staining along the glomerular capillary loops(GCL)continually and evenly in normotensive condition than in acute hypertension.,while in acute hypertensive condition,it stained intermittently and decreased obviously.At the same time,TRPC6 distributed in cytoplasm and nuclei of the podocytes in normal condition.The nucleoplasm of TRPC6 appeared marginated under acute hypertension.Conclusion:1.Abnomal hemodynamic conditions can reduce the expression of TRPC6.2.Abnomal hemodynamic conditions can cause TRPC6 increase in the cytoplasm and transfer to the cell nucleus.