Effects of inhibition of rhodopsin kinase of 4-aminopyridines compounds on epithelial-mesenchymal transition of rat peritoneal mesothelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Studies have confirmed that RhoA/ROCK signaling pathway plays an important role in the process of epithelial-mesenchymal transition. OBJECTIVE: To investigate the effect of Y-27632 (inhibition of rhodopsin kinase of 4-aminopyridines compounds) on the epithelial-mesenchymal transition of rat peritoneal mesothelial cells (RPMCs) induced by transforming growth factor ?1 (TGF-?1). DESIGN,TIME AND SETTING: An animal observational experiment was performed at the Laboratory of Nephrology in the First Affiliated Hospital of Sun Yat-sen University from March 2007 to March 2008. MATERIALS A total of ten healthy male Sprague Dawley rats were provided by the Animal Experimental Center of Sun Yat-sen University. Y-27632 was the product of Calbiochem Company (German). TGF-?1 was the product of R&D Company (USA). METHODS: Primary RPMCs were cultured in vitro. After synchronization for 24 hours, RPMCs were randomly divided into 4 groups control group (RPMCs cultured in the DMEM/F12 without serum), TGF-?1 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L TGF-?1), Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632), TGF-?1 +Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632. 2 hours later, 10 ?g/L TGF-?1 were added). The cells were collected 48 hours after culture. MAIN OUTCOME MEASURES: The mRNA and protein expression of E-cadherin and ?-Smooth muscle actin (?-SMA) were measured by RT-PCR. The protein expression level of E-cadherin, ?-SMA and vimentin was measured by Western blotting. RESULTS: Compared to control group, the expression of E-cadherin significantly decreased in the TGF-?1 group, and the expression of ?-SMA and vimentin significantly increased (P