ABSTRACT
Objective To observe the effect of fasudil on cytoskeleton reconstruction in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),as well as to study the protective mechanism of fasudil in the pathological changes of podocytes.Methods Conditionally immortalized mouse podocytes were treated with Ang Ⅱ (10-7 mol/L).The podocytes were pre-incubated for 30 min or 60 min with various concentrations of fasudil (10-8,10-7,10-5 mol/L),then Ang Ⅱ (10-7 mol/L) were added and furtherincubated for 24 hours.The cytoskeleton distribution of podocyte as indicated by F-actin and synaptopodin was observed by fluorescence microscopy and Western blotting.At the same time,the activity of Rho signal pathway that mediates actin filament polymerization was analyzed by measuring the extent of Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1) and myosin phosphatase-1 (MYPT1).Results Compared to the control group,AngⅡ disrupted the podocyte actin cytoskeleton and significantly decreased the expression of synaptopodin (P ＜ 0.05),while fasudil stabilized the actin filaments,and improved the synaptopodin expression (P ＜ 0.05).The expression enhancement of ROCK-1 and MYPT1 by Ang Ⅱ were reduced significantly by fasudil (P＜0.05).Conclusion The cytoskeleton reconstruction of podocytes can be induced by Ang Ⅱ and inhibited by fasudil,which suggests that the protective effect of fasudil may be partially contributed to the Rho/ROCK signaling pathway.