Study on Mechanism of 3-indolepropionic Acid Inhibiting EMT and Fibrosis in Peritoneal Mesothelial Cells
Objective To evaluate the effects of 3-indoleacetic acid(IPA)on epithelial-mesenchymal transition(EMT)and fi-brosis in mice peritoneal mesoepithelial cells stimulated by lipopolysaccharide.Methods Mouse peritoneal mesothelial cells be-fore and after LPS treatment were interfered with IPA at different concentrations(0.1,1.0 and 10 μmol/L),and the cell prolif-eration activity was examined by CCK-8 method to determine the optimal dose of IPA.Mice peritoneal mesothelium cells were divided into Control group,LPS group,LPS+IPA group,LPS+LY364947(TGF-β1 receptor inhibitor)group,LPS+IPA+LY364947 group and LPS+IPA+SRI-011381(TGF-β1 pathway activator)group.Cell proliferation viability was evaluated by CCK-8 assay and cell invasion was detected by Transwell assay.EMT-related factor(E-cadherin)and TGF-β1/Smad3 pathway-related factor(Smad3,p-Smad3)protein expression in cell supernatant was evaluated by Western blotting,and mesenchymal phe-notype α-smooth muscle actin(α-SMA)protein content was detected by ELISA.Results The optimal dose of IPA was 1.0μmol/L.Compared with Control group,cell proliferation and E-cadherin expression level were decreased(all P＜0.01).Invasivi-ty,α-SMA expression and p-Smad3/Smad3 were increased in LPS group(all P＜0.01).Compared with LPS group,cell prolifer-ative activity and E-cadherin expression level were increased(all P＜0.01),and invasivity,α-SM A expression and p-Smad3/Smad3 were decreased in LPS+IPA and LPS+LY364947 groups(all P＜0.01).Compared with LPS+IPA group,LPS+IPA+LY364947 group showed a more significant trend.The trends of measured indexes were reversed after SRI-011381 treatment in LPS+IPA group.Conclusion IPA inhibits Smad3 phosphorylation,thereby inhibiting cell EMT progression and alleviating LPS-induced peritoneal mesothelial cell injury.
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