Effect of TGF-β1 on epithelial-mesenchymal transition of rat peritoneal mesothelial cells and its mechanism / 中南大学学报(医学版)

ABSTRACT

Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epithelial-mesenchymal transition in rat peritoneal mesothelial cells(RPMCs) and its mechanism.Methods Primary peritoneal mesothelial cells of SP rats were cultured in vitro. After synchronization for 24 h, RPMCs were randomly divided into 2 groups Group A (control), Group B (TGF-β1, 10 μg/L). RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-cadherin, α-smooth muscle actin (α-SMA) and collagenⅠwere measured by RT-PCR and Western blot, respectively. The protein expression level of total RhoA was measured by Western blot. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, and assessed by Western blot. Results TGF-β1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of α-SMA and CollagenⅠ. TGF-β1 stimulation elicited a robust increase in RhoA activity in a time-dependent manner. RhoA activity peaked at 1 h.Conclusion RPMCs can be transdifferentiated into myofibroblast under the effect of TGF-(β1,)and the mechanism may be related to the activation of RhoA associated signal pathway.