Basic & Clinical Medicine ›› 2015, Vol. 35 ›› Issue (1): 48-53.

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Up-regulation of miR-200a attenuates TGF-β1-induced activation and collagen synthesis in rat pancreatic stellate cells

  

  • Received:2014-05-15 Revised:2014-07-23 Online:2015-01-05 Published:2014-12-30
  • Contact: Min XU E-mail:peterxu1974@163.com
  • Supported by:
    Natural Science Foundation of Jiangsu Province of China; The 169 fourth stage project for scientific research projects of Zhenjiang

Abstract: Objective To investigate the effect of miR-200a mimic on transforming growth factor β1-mediated activation and collagen secretion of rat pancreatic stellate cells. Methods PSCs were isolated and cultured from pancreatic tissue and identified by desmin, GFAP and α-SMA immunofluorescence staining. PSCs of 2nd generation were divided into control group, TGF-β1 group, TGF-β1+miR-NC group and TGF-β1+miR-200a mimic group. The levels of a-SMA and collagen I protein were measured by Western blot and immunofluorescence staining. The mRNA levels of a-SMA and collagenⅠ and the expression of miR-200a were detected by quantitative real-time PCR. Results TGF-β1 can stimulate the activation of PSCs and promote collagen synthesis in time-dependment manner (P<0.05). After transfection of the mimic, treating with the same concentration of TGF-β1, the expressions of protein and mRNA of both a-SMA and collagen I decreased significantly (P<0.01). Conclusion Over-expression of miR-200a significantly attenuates a-SMA activity and further affects the collagen synthesis of TGF-β1-dependent activation of PSCs. The mechanisms may inhibit the biological effects of TGF-β1.

Key words: TGF-β1, PSCs, miR-200a, fibrosis

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