CAJ | 학술논문

目的：研究实验性肝纤维化大鼠肝组织结缔组织生长因子（CTGF）启动子DNA甲基化变化及其对CTGF基因表达的影响。方法建立大鼠四氯化碳肝纤维化模型，以HE染色和Sirius red染色评估大鼠肝组织学变化；采用RT-PCR和Western blot法分别检测肝组织CTGF mRNA和蛋白质表达；采用亚硫酸氢盐修饰后测序评估肝组织CTGF基因DNA甲基化水平变化。结果在皮下注射四氯化碳6周时模型组大鼠肝组织呈III级纤维化改变；模型组大鼠肝组织CTGF mRNA及蛋白表达分别为（1.581±0.276）和（0.875±0.102），均显著高于正常对照组分别为（0.415±0.041）和（0.115±0.083），（P值均<0.01）；模型组CTGF基因启动子DNA甲基化率为（1.8±1.8）%，显著低于对照组（11.6±3.8）%，（P<0.01）。结论肝纤维化大鼠肝组织CTGF基因表达上调，其机制可能与CTGF基因DNA低甲基化改变相关。
목적：연구실험성간섬유화대서간조직결체조직생장인자（CTGF）계동자DNA갑기화변화급기대CTGF기인표체적영향。방법건립대서사록화탄간섬유화모형，이HE염색화Sirius red염색평고대서간조직학변화；채용RT-PCR화Western blot법분별검측간조직CTGF mRNA화단백질표체；채용아류산경염수식후측서평고간조직CTGF기인DNA갑기화수평변화。결과재피하주사사록화탄6주시모형조대서간조직정III급섬유화개변；모형조대서간조직CTGF mRNA급단백표체분별위（1.581±0.276）화（0.875±0.102），균현저고우정상대조조분별위（0.415±0.041）화（0.115±0.083），（P치균<0.01）；모형조CTGF기인계동자DNA갑기화솔위（1.8±1.8）%，현저저우대조조（11.6±3.8）%，（P<0.01）。결론간섬유화대서간조직CTGF기인표체상조，기궤제가능여CTGF기인DNA저갑기화개변상관。
Objective To investigate the change in DNA methylation of connective tissue growth factor(CT-GF) promoter and its effect on CTGF gene expression in experimental liver fibrosis in rats. Methods A rat model of liver fibrosis was established by subcutaneous injection of carbon tetrachloride for 6 weeks. The patho-logical changes in fibrotic livers were examed by HE and Sirius red staining; The levels of CTGF mRNA and protein expression in liver tissues were evaluated by reverse transcription polymerase chain reaction (RT-PCR)and Western blot,respectively;DNA methylation of CTGF gene was determined by sequencing after bisulfite modifica-tion. Results Liver fibrosis of grade III was observed after carbon tetrachloride injection at the end of 6 weeks. In liver tissues from model rats,the CTGF mRNA and protein were(1.581±0.276) and(0.875±0.102),respectively, which were significantly higher than those in the control rats(0.415±0.041)and(0.115±0.083),respectively,(P<0.01);Importantly,in liver tissues from models rats,the DNA methylation rate of CTGF gene promoter was(1.8±1.8)%, significantly lower than that in liver tissue from control rats (11.6±3.8)%,(P<0.01). Conclusion CTGF gene is upregulated in liver fibrosis,and this might be resulted from decreased DNA methylation in CTGF gene promoter.