The Effect Of The Complement C3a On The Podocyte Epithelial-Mesenchymal Transition And Its Mechanism In Adriamycin Nephropathy In Mice

Objective: To investigate the involvement of complement complement 3a(C3a)induce podocyte epithelial-mesenchymal transition and its mechanism with adriamycin nephropathy in mice.Methods: A total of 30 male BALB/c mice were randomized into control group,ariamycin nephrpathy group(ADR group),ariamycin+3mg/kg C3 a receptor antagonist(SB290157),ariamycin+10mg/kg SB290157,ariamycin+30mg/kg SB290157.On days 7,14 and 21 after the intervene,24-urine was collected from the mice to analyze the total urine proteins.The renal tissues were obtained on 21 days to observe the podocyte using electron microscopy;the deposition of C3 on the podocyte were examined by double immunohistochemistry;the expression of nephrin,podocin,?-SMA,FSP1,ILK,snail as well as podocyte number was measured by immunohistochemistry,quantification of nephrin,?-SMA,snail protein was carried out by western blot.Results: Compared with control group,the diffuse effacement of podocyte foot process was observed by electron microscopy in ADR group,the urine protein increased progressively in ADR group(P<0.05),the deposition of C3 on podocyte was examined by double immunohistochemistry significantly increase in ADR group(P<0.05),?-SMA,FSP-1,ILK,Snail,?-actinin-4 using immunohistochemistry increased progressively and nephrin,podocin significantly reduce in ADR group(P<0.05),the ?-SMA,snail proteins expression were increased and the nephrin podocin exprsssion reduced using western blot in ADR group(P<0.05).WT1 showed no significant reduction in each group(P>0.05).10mg/kg SB290157 can relieve the injure of podocyte foot process and significantly reduce 24-h urinary protein,the deposition of C3 and the expression of ?-SMA,FSP-1,ILK,Snail and ?-actinin-4 of podocyte and increase the expression of nephrin and podocin(P<0.05).But the diffuse effacement of podocyte foot process was observed by electron microscopy on the 3,30mg/kg SB290157 group and there was no statistical significance in the deposition of C3 and the expression of C3,?-SMA,FSP-1,ILK,Snail,?-actinin-4,nephrin,podocin compared with those in ADR group(P>0.05).Conclusion: 1.Podocyte undego epithelial-mesenchymal transition in adriamycin nephropathy in mice,which establish the animal model of epithelial-mesenchymal transition.2.There is the deposition of C3 on the podocyte in adriamycin nephropathy in mice,which indicate the important role of complement system on adriamycin induce the injury of podocyte.3.The C3 a receptor antagonist can suppress the process of podocyte epithelial-mesenchymal transition in adriamycin nephropathy in mice,which indicate complement C3 a can induce podocyte epithelial-mesenchymal transition in vivo.4.The expression of ILK and snail were increased in adriamycin nephropathy in mice and the C3 a receptor antagonist can reduce its expression,which indicate the important role of the ILK signing pathway in the process of complement C3 a induce podocyte epithelial-mesenchymal transition.5.The expression of ?-actinin-4 were increased in adriamycin nephropathy in mice and the C3 a receptor antagonist can reduce its expression,which indicate the important role of the cytoskeletal protein in the process of complement C3 a induce podocyte epithelial-mesenchymal transition.