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目的研究在肝纤维化发生发展中,结缔组织生长因子(CTGF)基因表达水平变化与肝纤维化病理分级和Ⅰ、Ⅲ型胶原基因表达的关系。方法以四氯化碳(CCl_4)皮下注射诱导大鼠肝纤维化作为实验模型,应用 HE 及 Sirius red 染色对肝纤维化进行病理分级,应用 Western blot 和(或)逆转录-聚合酶链反应(RT-PCR)检测在 CCl_4诱导2周和6周的肝纤维化大鼠与正常大鼠肝组织中 CTGF、Ⅰ及Ⅲ型胶原基因表达水平变化。结果 CCl_4皮下注射2周大鼠肝组织主要呈Ⅰ级纤维化病理改变,注射6周呈Ⅲ级纤维化病理改变;在Ⅰ级纤维化和Ⅲ级纤维化肝组织中,CTGF mRNA 和蛋白及Ⅰ、Ⅲ型胶原 mRNA 表达水平分别较正常对照上调0.64±0.078、2.66±0.269、1.83±0.096、2.85±0.096和2.81±0.055、6.41±0.224、3.25±0.13、2.69±0.082倍。结论 CTCF 基因表达水平变化与肝纤维化程度及Ⅰ型胶原基因表达水平变化呈显著正相关,与Ⅲ型胶原基因表达水平变化不相关,提示 CTGF 表达增高在肝纤维化发生发展中可能起重要作用,推测对CTGF 基因表达进行干预可能是防治肝纤维化的一个新策略。
Objective To study the relationship between the expression of connective tissue growth factor (CTGF) gene and the pathological grade of liver fibrosis and the expression of collagen type Ⅰ and Ⅲ in the development of hepatic fibrosis. Methods Liver fibrosis induced by subcutaneous injection of carbon tetrachloride (CCl_4) was used as an experimental model. Liver fibrosis was stained with HE and Sirius red stain. Western blot and / or reverse transcription - polymerase chain reaction RT-PCR) was used to detect the expression of CTGF, collagen type Ⅰ and type Ⅲ collagen mRNA in liver tissue of rats with hepatic fibrosis induced by CCl 4 for 2 weeks and 6 weeks respectively. Results The pathological changes of grade Ⅰ fibrosis were observed in the liver tissue of CCl_4 subcutaneously two weeks after injection of CCl_4, and the pathological changes of grade Ⅲ fibrosis were observed at 6 weeks after injection. In grade Ⅰ fibrosis and grade Ⅲ fibrosis, CTGF mRNA and protein, , And the mRNA expression of type Ⅲ collagen were up-regulated by 0.64 ± 0.078, 2.66 ± 0.269, 1.83 ± 0.096, 2.85 ± 0.096 and 2.81 ± 0.055, 6.41 ± 0.224, 3.25 ± 0.13 and 2.69 ± 0.082 times respectively. Conclusion The change of CTCF gene expression is positively correlated with the degree of hepatic fibrosis and the gene expression of type Ⅰ collagen, but not with the change of gene expression of type Ⅲ collagen. It suggests that the increased expression of CTGF may play an important role in the development of hepatic fibrosis It is speculated that intervention of CTGF gene expression may be a new strategy for prevention and treatment of hepatic fibrosis.