实用肝脏病杂志 ›› 2012, Vol. 15 ›› Issue (5): 430-433.doi: 10.3969/j.issn.1672-5069.2012.05.0019

• 基础研究 • 上一篇    下一篇

抑制结缔组织生长因子表达对肝星状细胞增殖和活化的影响*

彭梅娟, 郝春秋, 张野, 谢玉梅, 张岩, 白雪帆   

  1. 710038 西安市 第四军医大学唐都医院全军感染病诊疗中心
  • 收稿日期:2011-12-01 出版日期:2012-10-10 发布日期:2017-03-09
  • 通讯作者: 郝春秋,E-mail:haochq@fmmu.edu.cn
  • 作者简介:彭梅娟 女,33岁,主治医师。E-mail:pengmeijuan2007@yahoo.com.cn
  • 基金资助:
    国家自然科学基金(No:30571658)

Observation of inhibition of connective tissue growth factor by siRNA on the changes of proliferation and activation of hepatic stellate cells in vitro

Peng Meijuan, Hao Chunqiu, Zhang Ye, et al.   

  1. Center of Infectious Diseases of PLA,Tangdu Hospital,the Fourth Military Medical University,Xi’an 710038
  • Received:2011-12-01 Online:2012-10-10 Published:2017-03-09
  • Contact: Hao Chunqiu,E-mail: haochq@fmmu.edu.cn

摘要: 目的 观察抑制结缔组织生长因子(CTGF)表达对肝星状细胞系HSC-T6细胞周期、凋亡和增殖的影响。方法 构建含有靶向CGTF 小干扰RNA的重组慢病毒载体并感染HSC-T6细胞,应用MTT法检测细胞增殖情况;应用PI染色和流式细胞术检测细胞周期和凋亡。结果 抑制CTGF基因表达可抑制HSC-T6细胞增殖,同时显著增加HSC-T6细胞凋亡,凋亡细胞的百分比为17.58±5.81%,明显高于空白对照组(5.12±1.21%)和阴性对照组(1.37±0.31%);抑制CTGF基因表达可显著升高S期细胞百分比(53.24±4.47%),空白对照组和阴性对照组分别为44.80±4.701%和42.10±1.40%,而处于G2~M期细胞百分比(0.04±0.07%)则较空白对照组(9.03±0.45%)和阴性对照组(12.19±5.62%)显著降低,但抑制CTGF对G0~G1期细胞百分比无影响。结论 抑制CTGF可促进HSC-T6细胞凋亡,抑制其活化和增殖。抑制CTGF基因表达有可能作为抗肝纤维化的新靶点。

关键词: 肝星状细胞, 结缔组织生长因子, 细胞周期, 凋亡

Abstract: Objective To investigate the changes of cell cycle,apoptosis and proliferation of hepatic stellate cells(HSC)-T6 by inhibition of connective tissue growth factor(CTGF)by siRNA in vitro. Methods Recombinant lentiviruses containing siRNA targeted to GTGF were constructed and we let it infected HSC-T6 cells. Cell cycles and apoptosis were detected by flow cytometry with PI staining. Cell proliferation was measured by MTT. Results Suppression of GTGF induced the apoptosis of and reduced the proliferation of HSC-T6 cells; The percentage of apoptosis cells was 17.58 ± 5.81%,which was significantly higher than in blank control (5.12 ± 1.21%)and in negative control(1.37 ± 0.31%);The frequency of S cells was remarkably increased(53.24± 4.47%)with CTGF gene silence as compared with that(44.80± 4.701%)in blank control and that(42.10± 1.40%)in negative control;Meanwhile,the percentage of G2-M cells(0.04± 0.07%)was significantly decreased as compared with that(9.03± 0.45%)in blank control and that (12.19± 5.62%) in negative control. However,the G0-G1 cells were not influenced by inhibition of CTGF. Conclusion Inhibition of GTGF promotes the apoptosis and decreases the activation and proliferation of HSC-T6 cells. CTGF gene silence may serve as a new target to treat the liver fibrosis.

Key words: Hepatic stellate cells, Connective tissue growth factor, Cell cycle, Apoptosis