乙型肝炎肝纤维化组织差异蛋白的筛选及DDAH1的功能研究.docVIP

乙型肝炎肝纤维化组织差异蛋白的筛选及DDAH1的功能研究 　　作者：罗新华，程明亮，张权，杨勤 作者单位：1.贵阳医学院附院 感染科， 贵州 贵阳 550004; 2.贵阳医学院 病理生理学教研室， 贵州 贵阳 550504 　　【摘要】目的： 对比分析乙型肝炎纤维化和非纤维化肝组织的蛋白质组表达谱的差异，探讨二甲基精氨酸二甲基氨基水解酶1(DDAH1)在肝纤维化发生发展中的功能。方法： 利用双向凝胶电泳(2-DE)筛选肝纤维化和非纤维化肝组织差异表达的蛋白质，质谱鉴定差异蛋白质，Western-blot验证DDAH1的表达水平;测定肝组织中DDAH1活性，不对称二甲基精氨酸(ADMA)的含量，一氧化氮合酶(NOS)活性及NO2-/NO3-比值。结果： 鉴定出DDAH1等25个2倍以上的差异蛋白质;肝纤维化组织中DDAH1的活性降低， ADMA含量增加，NOS活性下降，一氧化氮(NO)含量减少。结论： 乙型肝炎肝纤维化与非纤维化肝组织的蛋白质表达谱有一定差异，DDAH1有可能通过调节ADMA的代谢而影响NO合成，参与了肝纤维化的发生发展。 　　【关键词】 肝炎，乙型，慢性; 纤维化; 蛋白质组学; 二甲基精氨酸; 二甲基氨基水解酶类 　　A Study on the Differential Proteins and Functions of DDAH1 in 　　Human Hepatitis B Derived Hepatic Fibrosis Tissues 　　LUO Xinhua, CHENG Mingliang, ZHANG Quan, YANG Qin 　　(1.Department of Infectious Disease, The Affiliated Hospital of Guiyang Medical College,Guiyang 550004, Guizhou, China; 　　2. Department of Pathophysiology, Guiyang Medical College, Guiyang 550004, Guizhou, China) 　　[Abstract] Objective: To compare and analyze the difference of proteomic expression profiles between human hepatitis B (HB) derived hepatic fibrosis tissues and non-fibrosis liver tissues, and analyze the functions of dimethyl aminohydrolase(DDAH1)in the formation and development of hepatic fibrosis. Methods: The differential expression of proteins were screened with two dimension electrophoresis (2-DE) and identified with mass spectrogram after the total protein was extracted from hepatic fibrosis tissue and nonfibrosis liver tissue. DDAH1 expression was verified with Western-blot. DDAH1 activity,asymmetry diethylarginine,(ADMA)concentration,nitricoxide synthase(NOS) activity,and nitrogen monoxidium(NO)concentration were determined. Results: Twenty-five proteins with 2-fold difference were identified. The results of DDAH1 by western blot showed conformity with that by 2-DE. Further study showed that, in comparing with those of non-fibrosis liver tissue, the activity of DDAH1 was lowered with the increase of ADMA in hepatic fibrosis tiss