CCl4诱导的大鼠肝纤维化模型肝纤维化逆转与MAPK信号通路的研究

Dynamic expression of MAPK signaling pathway on recovery hepatic fibrosis rats induced by CCl_4

目的通过检测CCl4诱导肝纤维化大鼠逆转恢复各时期MAPK信号通路蛋白动态变化,研究其在肝纤维化恢复期可能的调控作用。方法采用CCl4皮下注射SD大鼠12周,复制肝纤维化模型,分别于注射12周,停药后2、4、6、8周取材。通过检测各组ALT、AST、Hyp、LN、PⅢNP的含量变化和肝脏病理组织切片Masson染色,加以验证。采用West-ern blot法检测各组肝组织中MAPK通路蛋白表达。结果 ALT、AST、Hyp、LN、PⅢNP等反映肝损伤和纤维化进程的指标在第12周最高,即模型组最高,并且随着肝纤维化逆转而含量逐渐降低,Masson染色结果与之一致,表明大鼠肝纤维化模型期与各恢复期模型建立完成。p-ERK1/2在模型组表达明显降低,与正常组相比差异有显著性(P<0.01),恢复4周表达明显升高,与模型组相比差异有显著性(P<0.01),以后逐渐降低。p-JNK1/2在模型组表达明显降低(P<0.01),恢复期逐渐升高(P<0.05)。p-p38在模型组表达明显升高(P<0.01),恢复期逐渐降低(P<0.05)。p-ERK/ERK与Hyp相关系数r=-0.46,P=0.003;p-JNK/JNK与Hyp相关系数r=-0.53,P=0.0004;p-p38/p38与Hyp相关系数r=0.81,P=0.0001。结论 MAPK信号通路中的ERK、JNK和p38信号通路在肝纤维化逆转中发挥重要作用。

Aim To investigate the expression of MAPK signaling pathway in CCl4-induced recovery hepatic fibrosis rats and hence to explore their regulatory role.Methods SD rats were injected with CCl4 for 12 weeks to reproduce spontaneous reversal of liver fibrosis animal models.These models were divided into normal group,model group,2-week recovery,4-week recovery,6-week recovery and 8-week recovery according to the experimental design.ALT,AST,Hyp,LN,PⅢNP and Masson stainning of liver pathology of each group were verified.Western blot was used to detect the protein expression.Results The results of ALT,AST,Hyp,LN,PⅢNP and Masson stained tissue sections of liver pathology of each group suggested that animal model had been established.The expression of p-ERK1/2 was the lowest in the model group,with the normal group there were significant differences（P0.01）,4-week recovery was highest and significantly difference compared with the model group（P0.01）,then gradually reduced to normal level.p-JNK1/2 was the lowest in the model group（P0.01）,then increased in recovery groups（P0.05）.The expression of p-p38 was the highest in the model group（P0.01）,and gradually decreased in recovery groups（P0.05）.The coefficient correlation of p-ERK/ERK and Hyp r=-0.46,P=0.003,the coefficient correlation of p-JNK/JNK and Hyp r=-0.53,P=0.0004,and the coefficient correlation of p-p38/p38 and Hyp r=0.81,P=0.0005.Conclusion The MAPK signaling pathway may play a pivotal role in the spontaneous reversibility of hepatic fibrosis.