Title: The effect and mechanism of dexamethasone on AA induced epithelial-mesenchymal transition in HKC cells
Abstract: Objective To explore the effect and mechanism of dexamethasone(Dex)on AA induced epithelial-mesenchymal transition in HKC cells. Methods Cultured HKC cells were divided into five groups: negative control group, incubated with AA(10 μg/ml) alone group, incubated with Dex (100 μmol/L) alone group, incubated with AA(10 μg/ml) group and absolute ethanol(100 μmol/L)group, or incubated with AA (10 μg/ml) and Dex group at different concentrations (100, 10, 1, 0.1, 0.01 μmol/L) for 48 h. Confocal microscope was used to detect expressions of α-SMA and E-cadherin. The mRNA and protein expressions of α-SMA, E-cadherin, TGF-β1, p-Smad 3 and Smad 7 were assessed with real-time PCR and western Blot, respectively. In the other experiment, HKC cells were incubated with AA (10 μg/ml) in the absence or presence of Dex (100 μmol/L), respectively. The mRNA and protein expressions of α-SMA, E-cadherin, p-Smad 3, TGF-β1 and Smad 7 were assessed with real-time PCR and western Blot at different time points respectively. Results As revealed by confocal microscope, AA inhibited E-cadherin expression and enhanced α-SMA expression in HKC cells. Dex reversed AA-induced inhibition of E-cadherin and promotion of α-SMA expressions. Real-time PCR and western Blot showed that mRNA and protein expressions of α-SMA, p-Smad 3 and TGF-β1 significantly decreased, E-cadherin and Smad 7 significantly increased in cells incubated with AA and Dex, compared with that treated with AA alone (P 0.05). There was no significant difference between the group of AA alone and the group treated with AA and absolute ethanol for mRNA and protein expression of α-SMA, p-Smad 3 and TGF-β1 in HKC cells. Conclusions The results of the present study suggest that Dex may partially inhibit AA-induced EMT of HKC cells in vitro, which is probably contributed to the repression of TGF-β1 and p-Smad 3, up-regulation of Smad 7 expressions. Further study is needed.
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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