乙型肝炎肝纤维化组织差异蛋白筛选及DDAH1功能探究.docVIP

乙型肝炎肝纤维化组织差异蛋白筛选及DDAH1功能探究 　 作者：罗新华，程明亮，张权，杨勤 【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 　　［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 tissues (2.04±0.33 μmol/L vs 1.16±0.21 μmol/L, P＜0.01）, NOS activity lowered（4.39±0.66 U/ml vs 7.65±0.91 U/ml,P＜0.05）, and the NO content decreased（55.72±11.32 μmol/L vs 96.27±17.84 μmol/L,P＜0.01）. Conclusions: There is difference of protein expression spectra between HB derived hepatic fibrosis tissue and non-fibrosis liver tissue. DDAH1 might be involved in the formation and development of hepatic fibrosis by regulating ADMA metabolism and impacting NO synthesis. 　　［Key words］ hepatit