目的探讨大鼠肝纤维化组织中骨形成蛋白2(bone morphogenetic protein-2,BMP-2)的表达趋势及意义。方法用皮下注射四氯化碳构建大鼠肝纤维化模型,造模后4周、8周、12周分别检测血清层粘蛋白(LN)、透明质酸(HA)的变化,采用HE及Masson染色光学显微镜下观察肝组织损伤情况,采用Real—TimePCR、免疫组织化学和Western印迹检测各组大鼠肝组织中BMP-2的表达。采用单因素方差分析进行多组均数间的比较。结果肝纤维化模型组血清LN、HA明显升高。在对照组和肝纤维化模型组中BMP-2mRNA均有表达。模型组4周时BMP-2mRNA表达与对照组无显著差异(P〉0.05),模型组8周、12周较对照组显著下降(P均〈0.05)。与模型组4周相比,模型组BMP-2mRNA的表达在8周、12周显著下降(P均〈0.05),12周达最低。免疫组织化学证实,BMP-2在肝纤维化模型组较对照组大鼠肝组织中表达减少。在对照组和肝纤维化模型组中BMP-2蛋白均有表达。模型组4周时BMP-2蛋白表达与对照组无显著差异(P〉0.05),模型组8周、12周较对照组显著下降(P均〈0.05)。与模型组4周相比,模型组BMP-2蛋白的表达在8周、12周显著下降(P均〈0.05),12周达最低。结论BMP-2在正常SD大鼠肝脏有表达,随着肝纤维化进展,表达减少,提示BMP-2参与肝纤维化的发生、发展过程。
Objective To explore the expression and significance of bone morphogenetic protein-2 (BMP-2) in rats models of hepatic fibrosis. Methods SD rat models with hepatic fibrosis were established by injecting carbon tetrachloride(CC14). Serum laminin(LN) and hyaluronic acid(HA) levels were tested from hepatic fibrosis group at the time of 4 week, 8 week, 12 week. Pathological characters of liver tissue were observed under optical microscope after hema- toxylin-eosin and Masson staining. And the expressions of BMP-2 were detected by the methods of Real-Time poly- merase chain reaction (Real-Time PCR), immunohistochemistry and Western blot in rat fibrosis tissues. Comparisons between groups were analyzed by one-way ANOVA. Results LN and HA in model group were significantly higher than those in control group. All of normal control group and hepatic fibrosis groups expressed BMP-2 mRNA. Compared with control group, the expression of BMP-2 mRNA of the model group at week 4 had no significant difference, but were significantly lower at week 8 to 12. Compared with model group at week 4, the expression of BMP-2 mRNA of the model group at week 8 to 12 decreased significantly, and reached the nadir at week 12. Immunohistochemistry proved that the expressions of BMP-2 in hepatic fibrosis groups were all significantly lower than control group. All of normal control group and hepatic fibrosis groups expressed BMP-2 protein. Compared with control group, the expression of BMP-2 proteir, of the model group at week 4 had no significant difference, but were significantly lower at week 8 to 12. Compared with model group at week 4, the expression of BMP-2 protein of the model group at week 8 to 12 decreased significantly, and reached the nadir at week 12. Conclusion BMP-2 expressed in the liver of normal SD rats, and decreased along with the progress of hepatic fibrosis. These indicated that BMP-2 might be involed in the pathogenesis and development of hepatic fibrosis.