Objective: To explore the effect of transforming growth factor beta1 (TGF-beta1) 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-beta1, 10 mug/L). RPMCs were stimulated by 10 mug/L TGF-beta1 for different time. The mRNA and protein expression levels of E-cadherin, alpha-smooth muscle actin (alpha-SMA) and collagen I were 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-beta1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of alpha-SMA and CollagenI. TGF-beta1 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-beta1,and the mechanism may be related to the activation of RhoA associated signal pathway.