钙中性蛋白酶2及Bax在肝纤维化大鼠肝组织中的表达变化及意义  

Expression of calpain 2 and Bax in rat fibrotic liver tissues

作　　者：谢汝佳[1] 韩冰[1] 杨婷[1] 温静静[1] 杨勤[1] 

机构地区：[1]贵阳医学院病理生理教研室,贵州贵阳550004

出　　处：《中国病理生理杂志》2013年第9期1603-1608,共6页Chinese Journal of Pathophysiology

基　　金：贵州省科技厅国际科技合作项目(No.黔科合外G字[2010]7016号)

摘　　要：目的:探讨钙中性蛋白酶2(calpain 2)及Bcl-2相关X蛋白(Bax)在肝纤维化过程中的表达变化及其在肝纤维化过程中的可能作用。方法:雄性Wistar大鼠40只,随机分为正常对照4周、8周组和肝纤维化模型4周、8周组,每组各10只。肝纤维化组按3 mL/kg体重的剂量皮下注射40%CCl4植物油溶液,每隔3 d注射1次,造模时间分别为4周和8周;对照组大鼠皮下注射等量植物油溶液。采用TUNEL法检测肝组织中肝细胞凋亡情况;real-time PCR检测肝组织中calpain 2及bax mRNA的表达变化。免疫组织化学法及Western blotting检测肝组织中calpain 2及Bax蛋白的表达情况。结果:Real-time PCR检测发现肝纤维化4周和8周组大鼠肝组织中calpain2及bax mRNA表达较相应的正常对照组显著增加。免疫组化及Western blotting检测显示肝纤维化4周组大鼠肝组织中calpain 2蛋白的表达与正常4周组比较无显著差异;随着肝纤维化程度的加重,肝纤维化8周时大鼠肝组织中calpain 2的表达显著增加;而肝组织中Bax的表达从肝纤维化4周时就显著增加,肝纤维化8周时达到高峰。此外,通过TUNEL法检测发现肝纤维化4周和8周组大鼠肝组织中肝细胞凋亡的数目较正常组大鼠显著增加。结论:Calpain 2与Bax可能参与了肝纤维化的发生发展过程。AIM：To observe the expression of calpain 2 and Bcl-2-associated X protein （Bax） in rat fibrotic liver tissues and to explore their effects on hepatic fibrosis. METHODS：Forty male Wistar rats were randomly divided into four groups （each n=10）： 4-week control group, 8-week control group, 4-week liver fibrosis group and 8-week liver fibrosis group. Liver fibrosis model was induced by subcutaneous injection of 40% CCl4 （3 mL/kg） every 3 days for 4 or 8 weeks. The apoptosis of hepatocytes was detected by TUNEL. Additionally, the mRNA expression of calpain 2 and bax was determined by real-time PCR, and the protein expression of calpain 2 and Bax was determined by immunohistochemistry and Western blotting. RESULTS：Real-time PCR showed that the mRNA expression of calpain 2 and bax in liver tissues was elevated in 4-week and 8-week liver fibrosis groups. The results of immunohistochemistry and Western blotting revealed that there was no difference of calpain 2 protein expression in liver tissues between 4-week liver fibrosis group and control group, but that in 8-week liver fibrosis group was obviously increased. The protein expression of Bax in 4-week and 8-week liver fibrosis groups was higher than that in control groups. Additionally, the numbers of apoptotic hepatocytes in 4-week and 8-week liver fibrosis groups were obviously increased compared with control groups.CONCLUSION：Calpain 2 and Bax may play important roles in the process of liver fibrosis.

关 键 词：肝纤维化 钙中性蛋白酶2 Bcl-2相关X蛋白 

分 类 号：R363[医药卫生—病理学]