摘要
目的探讨结缔组织生长因子(CTGF)在肝纤维化过程中的表达变化,并观察其对大鼠肝星状细胞(HSC)功能的影响。方法采用ABC免疫组化法观察肝纤维化动物模型制模后不同时间点的CTGF表达变化;构建CTGF 2个不同基因位点的shRNA重组体质粒并转染HSC,利用实时PCR检测CTGF表达,通过MTT及细胞粘附实验检测细胞功能变化。结果随着肝纤维化进展,CTGF阳性表达呈增强趋势(P<0.05);成功构建表达靶向CTGF的shRNA重组真核表达质粒;MTT实验结果显示干扰质粒组细胞增殖水平[吸光度值为(0.81±0.06)]弱于对照组(1.39±0.07)及阴性质粒组(1.28±0.06),均P<0.05;细胞粘附实验表明干扰质粒组细胞粘附率为(38.1±6.2)%,明显低于对照组的(63.1±3.6)%和阴性质粒组的(62.2±5.0)%,均P<0.05。结论 CTGF参与了肝纤维化的过程,是促进肝纤维化发展的最重要细胞因子之一,且表达水平随肝纤维化进展逐渐升高。CTGF shRNA重组体能明显抑制HSC的增殖和粘附能力。
Objective To investigate the expression of connective tissue growth factor（CTGF）in rat hepatic fibrosis and as-sess its effects on hepatic stellate cells（HSC-T6）.Methods The rat models of hepatic fibrosis were established by hypodermic inj ection of CCl4.The expression of CTGF in the liver was detected by immunohistochemistry at different time after the model establishment.shRNA expression plasmid vectors were constructed,targeting two different gene loci of CTGF.MTT assay and adhesion assay were conducted to examine the changes of cell functions after transfection of CTGF shRNA into HSC-T6 cells.Results The expression of CTGF in the liver increased with the development of hepatic fibrosis（P〈0.05）.Transfection of CTGF shRNA caused a significant decrease in the proliferation and adhesion of HSC-T6 cells[absorbance（A）value of prolif-eration：（0.81±0.06）for interference plasmid group vs.（1.39±0.07）,（1.28±0.06）for control and negative plasmid groups, respectively,P〈0.05;adhesion rate：（38.1±6.2%）for interference plasmid group vs.（63.1±3.6）%,（62.2±5.0）% for con-trol and negative plasmid groups,respectively,P〈0.05].Conclusion CTGF is implicated in the development of hepatic fibro-sis.CTGF shRNA recombinants can significantly suppress the proliferation and adhesion of hepatic stellate cells.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2016年第3期284-287,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81402640)
湖北省自然科学基金资助项目(No.2014CFB406)
武汉市卫生计生委科研项目(No.WX15B23)
武汉市中青年医学骨干人才培养工程[武卫生计生(2014)77号]
关键词
结缔组织生长因子
肝纤维化
肝星状细胞
细胞增殖
细胞粘附
connective tissue growth factor
hepatic fibrosis
hepatic stellate cell
cell proliferation
cell adhesion
