中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (40): 7533-7538.doi: 10.3969/j.issn.2095-4344.2012.40.023

• 器官移植基础实验 basic experiments of organ transplantation • 上一篇    下一篇

川芎嗪和siRNA沉默转化生长因子β1干预大鼠肝星状细胞 结缔组织生长因子的表达

向 颖1，李 军2，贾 茜1，阳 宏1，李孝生1   

  1. 1重庆医科大学附属第二医院消化内科，重庆市 400010
    2重庆医科大学附属口腔医院牙体牙髓科，重庆市 401147
  • 收稿日期:2012-02-25 修回日期:2012-04-19 出版日期:2012-09-30 发布日期:2012-09-30
  • 通讯作者: 李孝生，硕士，教授，硕士生导师，重庆医科大学附属第二医院消化内科，重庆市 400010 LXS6896@126. com
  • 作者简介:向颖★，女，1984年生，重庆市人，重庆医科大学在读硕士，主要从事肝纤维化的防治研究。 xiangyinga@ 126.com

Effects of tetramethylpyrazine and silencing transforming growth factor-beta 1 by small interfering RNA on expression of connective tissue growth factor of rat hepatic stellate cells

Xiang Ying1, Li Jun2, Jia Qian1, Yang Hong1, Li Xiao-sheng1   

  1. 1Department of Gastroenterology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
    2Department of Endodontics, Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing 401147, China
  • Received:2012-02-25 Revised:2012-04-19 Online:2012-09-30 Published:2012-09-30
  • Contact: Li Xiao-sheng, Master, Professor, Master’s supervisor, Department of Gastroenterology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China LXS6896@126.com
  • About author:Xiang Ying★, Studying for master’s degree, Department of Gastroenterology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China xiangyinga@126.com

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摘要：

背景：作者前期研究发现川芎嗪可以通过抑制肝星状细胞的增殖、阻抑p38MARK信号通路、下调结缔组织生长因子的表达等多途径发挥抗肝纤维化的作用。一般认为转化生长因子β1是促进纤维化发生最重要的细胞因子，结缔组织生长因子是转化生长因子β1的下游效应介质，siRNA靶向抑制转化生长因子β1可能成为治疗肝纤维化的新方法。
目的：观察川芎嗪及化学合成的siRNA转化生长因子β1对大鼠肝星状细胞结缔组织生长因子表达的影响。
方法：体外培养大鼠肝星状细胞，从3条siRNA转化生长因子β1中筛选一条有效的基因沉默片段，然后利用脂质体转染试剂共同转染培养24 h的细胞作为转染组，并设计空白对照组、转染试剂组、川芎嗪组及联合治疗组。
结果与结论：与空白对照组相比，转染组、川芎嗪组、联合治疗组Ⅰ、Ⅲ型胶原及结缔组织生长因子mRNA表达均下调(P < 0.05)。在转染组、川芎嗪组及联合治疗组，结缔组织生长因子蛋白表达均下调(P < 0.05)；培养上清液中Ⅰ型胶原和Ⅲ型胶原含量均降低(P < 0.05)。结果表明，siRNA 沉默转化生长因子β1基因能下调结缔组织生长因子的表达，减少Ⅰ、Ⅲ型胶原的表达。川芎嗪也能下调结缔组织生长因子的表达，但对Ⅰ、Ⅲ型胶原表达的抑制作用更加显著，提示川芎嗪抗肝纤维化可能是多种作用靶点。

关键词: 川芎嗪, 小干扰RNA, 转化生长因子β1, 肝星状细胞, 结缔组织生长因子

Abstract:

BACKGROUND: Our previous studies have demonstrated that tetramethylpyrazine can anti-hepatic fibrosis via inhibiting hepatic stellate cells proliferation, inhibiting p38 mitogen-activated protein kinase signal pathway, as well as down-regulating the expression of connective tissue growth factor, and so on. Generally, transforming growth factor-β1 is the foremost cell factor in promoting hepatic fibrosis, and connective tissue growth factor is the downstream effect medium of transforming growth factor-β1, small interfering RNA targeting transforming growth factor-β1 can be a new way in the treatment of hepatic fibrosis.
OBJECTIVE: To investigate the effect of tetramethylpyrazine and chemosynthesis small interfering RNA transforming growth factor-β1 on the expression of connective tissue growth factor of rat hepatic stellate cells.
METHODS: Hepatic stellate cells were cultured in vitro, an effective segment of gene silencing was screened out from three segments of small interfering RNA transforming growth factor-β1, then it was transfection group utilizing lipsome transfection reagent to transfect together the cells which have been cultured for 24 hours, and blank control group, transfection reagent group, tetramethylpyrazine group, and union treatment group were designed.
RESULTS AND CONCLUSION: Compared with blank control group, the expression of collagen type Ⅰ, collagen type Ⅲ and connective tissue growth factor mRNA were reduced in transfection group, tetramethylpyrazine group and union treatment group (P < 0.05). The connective tissue growth factor protein expression was down-regulated in transfection group, tetramethylpyrazine group and union treatment group (P < 0.05); contents of collagen type Ⅰ and collagen type Ⅲ in the supernatant were reduced (P < 0.05). Result show that the small interfering RNA silencing transforming growth factor-β1 can reduce the expression of collagen type Ⅰ and collagen type Ⅲ and down-regulate the expression of connective tissue growth factor. The expression of connective tissue growth factor can be down-regulated by tetramethylpyrazine, but it is more pronounced to the inhibiting expression of collagen type Ⅰ and collagen type Ⅲ, cluing that the anti-hepatic fibrosis of tetramethylpyrazine may be multifarious action targets.

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