CAJ | 학술논문

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4 d、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Epac1、Epac2及TGFβl 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Epac1(0.031 28±0.008 96)和Epac2(0.034 43±0.002 45)蛋白主要表达于肝细胞胞质.Epac1在造模后4 d(0.023 97±0.003 81)、1周(0.015 81±0.002 48)表达下降,2周后开始升高,6周时达高峰,为0.039 54±0.001 43,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0.011 21±0.001 32,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0.011 30±0.001 03,与对照组的0.002 08±0.000 18相比,差异有统计学意义(t=-23.36,P＜0.05).Epac1、Epac2和TGFβ1 mRNA的变化趋势与蛋白水平变化基本一致.相关分析显示,Epac1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(rs=-0.409,P＜0.05).结论 Epac1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.
목적 관찰Epac재실험성대서간섬유화과정중적동태변화.방법 42지SD웅성대서분위대조조6지,모형조36지,모형조재안4 d、1주、2주、4주、6주、8주분위6개아조.채용복강주사이갑기아초알건립대서간섬유화모형.채용HE화Masson염색관찰간조직병이학변화,RT-PCR、면역조직화학화Western인적법분별검측Epac1、Epac2급TGFβl mRNA화단백재조모과정중적동태변화급간조직중적정위.통계학처리채용단인소방차분석、LSD-t검험、Dunnett T3검험화Pearson직선상관분석.결과 성공건립대서간섬유화모형.대조조간조직Epac1(0.031 28±0.008 96)화Epac2(0.034 43±0.002 45)단백주요표체우간세포포질.Epac1재조모후4 d(0.023 97±0.003 81)、1주(0.015 81±0.002 48)표체하강,2주후개시승고,6주시체고봉,위0.039 54±0.001 43,여대조조상비,차이유통계학의의(t=5.47、11.58、-6.18,균P＜0.05).Epac2표체수평재모형조균하강,4주시체최저수평,위0.011 21±0.001 32,여대조조상비,차이유통계학의의(t=24.50,P＜0.05).TGFβl재모형조표체증가,4주시최명현,위0.011 30±0.001 03,여대조조적0.002 08±0.000 18상비,차이유통계학의의(t=-23.36,P＜0.05).Epac1、Epac2화TGFβ1 mRNA적변화추세여단백수평변화기본일치.상관분석현시,Epac1단백수평여간섬유화병정정정상관(rs=0.703,P＜0.01),이Epac2단백수평여간섬유화병정정부상관(rs=-0.409,P＜0.05).결론 Epac1재간섬유화형성과정중정선하강후승고추세,Epac2정지속하강추세;Epac가능삼여료간섬유화적발생발전과정.
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 Epac1, Epac2 and transforming gronth factor (TGF)β1 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 was established successfully. In control group, Epac1 (0. 031 28±0. 008 96) and Epac2 protein (0.034 43±0. 002 45) mainly expressed in the cytoplasm of hepatocytes. In model group, the level of Epac1 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±0.001 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 Epac1, Epac2 and TGFβ1 mRNA were consistent with the trend of protein levels.Correlation analysis showed that Epac1 protein was positively correlated with the course of HF (r =0. 703, P＜0.01 ), while Epac2 protein was negatively correlated (r = - 0. 409, P＜0.05). Conclusions During the progression of HF, Epac1 expression tends to decrease firstly and increase afterwards,while Epac2 expression declines continually. Epac may be involved in the pathogenesis of HF.