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  • Title: [Effects of pioglitazone on the transdifferentiation and CTGF expression of renal tubular epithelial-myofibroblast in vitro].
    Author: Pu L, Fan JM.
    Journal: Sichuan Da Xue Xue Bao Yi Xue Ban; 2010 Jan; 41(1):15-9. PubMed ID: 20369462.
    Abstract:
    OBJECTIVE: To investigate whether pioglitazone can inhibit TGF-beta-induced phenotypic remodeling of renal tubular epithelial cell and the expression of CTGF, so to illuminate the anti-fibrotic mechanism of pioglitazone. METHODS: Cultured NRK52E cells were divided into control group, TGF-beta-treated (3 ng/mL) group, TGF-beta (3 ng/mL)and DMSO-treated group, TGF-beta and pioglitazone-treated groups with different doses. The morphological change of cultured tubular cells was observed under phase-contrast microscopy. Immunohistochemistry were used to assess the alpha-SMA+ cells, and the expression of alpha-SMA mRNA, CTGF mRNA was measured by RT-PCR. The level of collagen type III in the culture supernatant was measured by ELISA. RESULTS: 1) Confluent NRK52E cells cultured in medium with 5% FCS showed a normal epithelial morphological state and cobblestone pattern. All cells almost had no expression of alpha-SMA and CTGF. After the NRK52E cells were cultured with TGF-beta 3 ng/mL for 5 days, many cells had strong cytoplasmic staining for alpha-SMA, and alpha-SMA+ cells showed marked hypertrophy and elongated shape. The levels of alpha-SMA mRNA and CTGF mRNA in TGF-beta-treated group cells were higher than those of normal control group (P < 0.05). Compared with control group, there were significant increases of collagen type III content in supernatant of TGF-beta-treated groups (P < 0.05). 2) The phenotypic changes induced by TGF-beta were prevented by incubation with pioglitazone. Pioglitazone inhibited the expression of alpha-SMA mRNA and CTGF mRNA of NRK52E cells, and the supernatant content of collagen type III in a dose-dependent manner (P < 0.05). 3) CTGF mRNA expression of NRK52E cells was significantly associated with alpha-SMA mRNA expression (r = 0.975, P< 0.01), and the content of collagen type III (r = 0.988, P<0. 01). The expression of rSMA mRNA was correlated with the content of collagen type IT (r 0. 952, P<0. 01). CONCLUSION: Pioglitazone may negatively regulate TEMT through inhibiting the expression of CTGF.
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