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CJAP ›› 2018, Vol. 34 ›› Issue (2): 122-125.doi: 10.12047/j.cjap.5568.2018.030

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Effects of centellaasiatica granule on the expression of Smad 2/3, Smad 7 and collagen Ⅳ in the mesangial cells stably expressed TGF-β1

MA Ji-wei1, WANG Hong-tian1, LIU Hao-fei1, DONG Lei-peng1, DING Yuan2, BAI Ji-qiong3, ZHANG Zhu1, DONG Li-jie4   

  1. 1. Department of Nephrology, Henan University of Traditional Chinese Medicine Affiliated First Hospital, Zhengzhou 450000;
    2. Xinyang Central Hospital, Xinyang 464000;
    3. Gongyi City People's Hospital, Gongyi 451200;
    4. Tianjin Medical University Eye Hospital, Eye Institute, Tianjin 300384, China
  • Received:2017-06-23 Revised:2017-10-30 Online:2018-03-28 Published:2018-05-22
  • Supported by:
    国家自然科学基金支持项目(81570872);天津市应用基础与前沿技术研究计划一般项目(15JCYBJC24900)

Abstract: Objective:Stably expressed transforming growth factor -beta 1(TGF-β1)MCs were obtained and the effects of centellaasiatica (CA) granule on the expressions of Smad 2/3, Smad 7 and collagen Ⅳ and the level of Smad 2/3 phosphorylation were observed. Methods:Lipofectin method was used to transfect TGF-β1 vector into MC, and the stably expressed TGF-β1 cell lines were selected by G418. The cells were divided into three groups. Control group:normal MC + RPMI 1640 + 10% normal rat serum; TGF-β1 group:stably expressed TGF-β1 MC + RPMI 1640 + 10% normal rat serum; CA group:stably expressed TGF-β1 MC + RPMI 1640 + 10% rat serum containing high CA. The experiments were repeated for five times. The contents of TGF-β1 and collagen Ⅳ in the culture medium were detected with ELISA, the expressions of mRNA and protein of TGF-β1, Smad 2/3, Smad 7 and the level of Smad 2/3 phosphorylation were detected by using real time quantitative polymerase chain reaction and Western blot. Results:The contents of TGF-β1 and collagen Ⅳ in the culture medium of stably-expressed TGF-β1 MC were increased significantly, and the CA could reverse the effects of TGF-β1. The expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation were increased significantly in TGF-β1 transfected MC, and CA could dramatically reduce the expressions of mRNA and protein of TGF-β1, Smad 2/3 and the level of Smad 2/3 phosphorylation. The high expression of TGF-β1 decreased the expression of Smad 7 mRNA and protein, and the CA could antagonize the effect of mRNA expression. Conclusion:The MCs stably-expressed TGF-β1 can activate the TGF-β1/Smad signal pathway and increase the expression of collagen Ⅳ. CA can decrease the occurrence of diabetic nephropathy(DN) by reducing the production of collagen Ⅳ through inhibiting the TGF-β1/Smad signal pathway.

Key words: rat, diabetic nephropathy, centellaasiatica, transforming growth factor-beta 1, Smad signaling pathway, collagen Ⅳ

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