结缔组织生长因子在实验性肝纤维化中的表达

Connective tissue growth factor expression in experimental hepatic fibrosis.

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 investigate the changes of connective tissue growth factor(CTGF)gene expression level and the association with pathological staging of liver fibrosis and changes of type Ⅰ,Ⅲ collagen gene expression levels in he- patic fibrogenesis.Methods Hepatic fibrosis rat models were created by subcutaneous injections of CCl_4,and the de- gree of hepatic fibrosis in model rats was determined by HE staining and Sirius red staining.The expressions of CTGF, type Ⅰ and Ⅲ collagen gene of the rat liver tissue specimens were analysed with the Western blot and/or Reverse tran- scription polymerase chain reaction(RT-PCR).Results Grade I fibrosis was observed in rat liver tissue of CCl_4 in- ductiun for 2 weeks,and grade Ⅲ fibrosis in that of CCl_4 induction for 6 weeks.The CTGF mRNA and protein and type Ⅰ and Ⅲ collagen mRNAs in hepatic tissue of grade Ⅰ fibrosis and grad Ⅲ fibrosis were increased 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、2.69±0.082 fold com- pared with normal control,respectively.Conclusion The changes of CTGF gene expression level significantly correla- ted with the degree of liver fibrosis and type Ⅰ collagen gene expression,but not with type Ⅲ collagen expression in liver fibrogenesis.These findings suggested that the upregulation of CTGF gene expression might play an important role in the process of liver fibrogenesis,and postulating the interference of CTGF expression might be a new strategy.