细胞增殖与凋亡相关蛋白质在大鼠肝纤维化形成与消减中的动态变化  

Proteomic analysis of proliferation and apoptosis in carbon tetrachloride induced rat liver flbrosis

作　　者：刘莺[1] 刘平[1] 刘成海[1] 胡义杨[1] 徐列明[1] 慕永平[1] 都广礼[1] 

机构地区：[1]上海中医药大学科技实验中心,201203

出　　处：《中华肝脏病杂志》2005年第8期563-566,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金重大专项(90409020)国家自然科学基金(30371835)

摘　　要：目的通过对四氯化碳诱导纤维化大鼠肝组织蛋白质组的分析,研究与增殖、凋亡相关蛋白质在肝纤维化过程中的动态变化,探讨细胞增殖与凋亡在肝纤维化形成及发展中的作用。方法将Wistar 大鼠随机分为正常组、模型组。模型组大鼠皮下注射40%四氯化碳-橄榄油溶液,每周2次,共12周,停止刺激后再观察4周。分别于第4、8、12、16周末分批取材。留取肝组织分别做病理组织学、羟脯氨酸检查与蛋白质的分步提取。蛋白质定量后进行二维电泳,凝胶银染,用PDQUEST 2-DE图像分析软件对获得的蛋白质图谱进行分析,运用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定30多个差异表达的蛋白质。结果从第1周开始模型组大鼠肝组织的胶原沉积、羟脯氨酸持续增高,在12周最高(P<0.05),16周下降(P<0.05),各时间点模型组与正常组比较差异有统计学意义(P<0.05);模型组大鼠肝组织蛋白质组较正常组也有较大改变。经MALDI-TOF-MS鉴定与细胞增生相关的蛋白质或酶为:细胞增殖核抗原p120与p40、细胞周期蛋白F和泛素结合酶UBC7;与细胞凋亡相关的蛋白质主要有胱门蛋白酶-12,只在模型组大鼠肝组织表达。结论与细胞增生、凋亡的相关蛋白质表达量在肝纤维化发生发展的不同阶段呈动态变化;在肝纤维化形成过程中细胞增生与凋亡受到相关蛋白质的调节。Objective To study the proliferation and apoptosis in carbon tetrachloride induced rat liver fibrosis.Methods Wistar rats were injected subcutaneously with 40% CCl4-olive oil twice weekly for 12 weeks. Liver tissues were obtained at the end of 4, 8, 12 and 16 weeks for histological examination, hydroxyproline （Hyp） assay and proteomic analysis. After two dimension electrophoresis （2-DE）, the silver stained gels were analyzed with PDQUEST 2-DE. More than 30 differentially expressed proteins were identified by MALDI-TOF-MS. Results The degree of collagen deposition and hydroxyproline content of the fibrotic livers increased continuously during the 12 weeks of CCl4 administration, peaked at the end of week 12 （P〈0.05） and declined significantly at week 16 （P〈0.05）. Significant differences were observed in two parameters at each time point between the control and the model group. Meanwhile,dramatic change of hepatic proteome in the model group rats was also seen. Differentially expressed proteins identified by MALDI-TOF-MS were categorized as proliferation-related proteins/enzymes （proliferating cell nuclear antigen p120,p40 and cyclin F ubiquitin-conjugating enzyme 7 UBC7）, and apoptosis-related proteins, mainly caspase-12 which was absent in the control rats. Conclusion Proliferation and apoptosis related proteins are expressed dynamically in different stages of rat liver fibrosis induced by CCl4.

关 键 词：细胞增殖 细胞凋亡 蛋白质 大鼠 肝纤维化 病理组织学 

分 类 号：R575.2[医药卫生—消化系统]