Epac在大鼠肝纤维化模型中的动态变化  

Dynamic expressions of exchange protein directly activated by cyclic adenosine monophosphate in ratmodel of liver fibrosis

作　　者：戴志娟[1] 陈永平[1] 程瑗[1] 叶超[1] 金晓芝[1] 林镯[1] 张磊[1] 谷甸娜[1] 

机构地区：[1]温州医学院附属第一医院感染内科

出　　处：《中华传染病杂志》2011年第1期11-17,共7页Chinese Journal of Infectious Diseases

基　　金：王宝思肝纤维化研究基金资助项目（20080017）;浙江省自然科学基金资助项目（Y207464）;温州市科技局项目（H20090014,Y20090269）;浙江省教育厅项目（Y201009942）

摘　　要：目的观察Epac在实验性大鼠肝纤维化过程中的动态变化。方法42只SD雄性大鼠分为对照组6只，模型组36只，模型组再按4d、1周、2周、4周、6周、8周分为6个亚组。采用腹腔注射二甲基亚硝胺建立大鼠肝纤维化模型。采用HE和Masson染色观察肝组织病理学变化，RT-PCR、免疫组织化学和Western印迹法分别检测Epacl、Epac2及TGFβI mRNA和蛋白在造模过程中的动态变化及肝组织中的定位。统计学处理采用单因素方差分析、LSD-t检验、Dunnett T3检验和Pearson直线相关分析。结果成功建立大鼠肝纤维化模型。对照组肝组织Epacl（0．03128±0．00896）和Epac2（O．03443±0．00245）蛋白主要表达于肝细胞胞质。Epacl在造模后4d（O．02397±0．00381）、1周（O．01581士0．00248）表达下降，2周后开始升高，6周时达高峰，为0．03954±0．00143，与对照组相比，差异有统计学意义（t-5．47、11．58、-6．18，均P〈0．05）。Epac2表达水平在模型组均下降，4周时达最低水平，为0．01121±0．00132，与对照组相比，差异有统计学意义（T=24．50，P％0．05）。TGFβl在模型组表达增加，4周时最明显，为0．01130±0．00103，与对照组的0．00208±0．00018相比，差异有统计学意义（t=23．36，P〈O．05）。Epacl、Epac2和TG即1mRNA的变化趋势与蛋白水平变化基本一致。相关分析显示，Epacl蛋白水平与肝纤维化病程呈正相关（rs=0．703，P〈0．01），而Epac2蛋白水平与肝纤维化病程呈负相关（rx=0．409，P〈O．05）。结论Epacl在肝纤维化形成过程中呈先下降后升高趋势，Epac2呈持续下降趋势；Epac可能参与了肝纤维化的发生发展过程。Objective To investigate the dynamic expressions of exchange protein directly activated by cyclic adenosine monophosphate （cAMP） （Epac） in rat model of hepatic fibrosis（HF）. Methods Forty two male SD rats were divided into control group （n =6） and model group （n= 36） which was divided into six subgroups of day 4, week 1, week 2, week 4,week 6 and week 8 with six rats in each subgroup. The rat model of HF was established by intraperitoneal injection of dimethylnitrosamine （DMN）. The pathological changes of liver were observed by Hematoxylin-Eosin and Masson staining. Reverse transcription-polymerase chain reaction （RT-PCR）, immunohistochemistry and Western blot were employed to detect the mRNA and protein expressions of Epacl, Epac2 and transforming gronth factor （TGF） [31 during the process of modeling and localization in the liver. The statistical analysis was done using one-factor ANOVA, LSD-t test, Dunnett T3 test and Pearson linear correlation analysis. Results Rat model of liver fibrosis wasestablished successfully. In control group, Epacl （0. 031 28i0. 008 96） and Epac2 protein （0. 034 43 ~0. 002 45） mainly expressed in the cytoplasm of hepatocytes. In model group, the level of Epacl decreased at day 4 （0. 023 97~0. 003 81） and week 1 （0. 015 81±0. 002 48） ,then began to increase at week 2 of modeling and peaked at week 6 （0. 039 54~0. 001 43）, which had statistical significance compared to the control group （t^5. 47,11. 58 and --6.18, respectively; all P〈~0.05）. Epac2 protein expression declined after modeling, reached the lowest level at week 4 （0. 011 21±0.001 32）, which had statistical significance compared to the control group （t=24. 50, P〈0. 05）. TGFβ1 protein expression increased after modeling and peaked at week 4 （0. 011 30±0.00l 03） which had statistical significance （t=-23. 36, P〈0. 05） compared to the control group （0. 002 08± 0. 000 18）. The expressions of Epacl, Epac2 and TGFβ1 mRNA were consistent with the

关 键 词：二甲基亚硝胺 肝纤维化 Epact转化生长因子βl 疾病模型 动物 

分 类 号：R575.2[医药卫生—消化系统]