中文摘要：

目的探讨脂酰肌3激酶-蛋白激酶B-雷帕霉素靶蛋白（PI3K/AKT/mTOR）在华支睾吸虫感染小鼠致肝纤维化作用。方法 6周龄雌性C3H/HeN小鼠随机分为正常对照组和华支睾吸虫感染组,感染组每只小鼠经口灌胃45个华支睾吸虫囊蚴,对照组每只小鼠经口灌入等量的生理盐水,感染28d后取出小鼠肝脏组织,Masson染色,观察病理变化;免疫组织化学染色检测肝脏组织中α-SMA和p-mTOR蛋白的定位与表达;实时荧光定量PCR（qRT-PCR）检测小鼠肝脏组织中α-SMA、COL1和TGF-βmRNA表达水平;Western blot法检测小鼠肝脏组织中TGF-β、α-SMA、pPI3K、p-AKT和p-mTOR的蛋白水平。结果成功建立小鼠肝纤维化模型。与对照组相比,感染组小鼠肝脏的胶原沉积显著增加,α-SMA及p-mTOR阳性细胞数量显著增多（t=-8.374,P〈0.01;t=-9.545,P〈0.01）,α-SMA、COL1、TGF-βmRNA（t=-3.553,P〈0.01;t=-3.448,P〈0.01;t=-3.052,P〈0.01）以及α-SMA、TGF-β、p-PI3K、p-AKT和p-mTOR的蛋白表达水平显著升高（t=-7.076,P〈0.01;t=-5.054,P〈0.01;t=-6.941,P〈0.01;t=-8.123,P〈0.01;t=-7.445,P〈0.01）。结论 PI3K/AKT/mTOR在华支睾吸虫感染小鼠肝脏高度表达,因此推测该信号通路在华支睾吸虫感染致肝纤维化中发挥重要作用。

英文摘要：

Objective To investigate the potential roles of the PI3K/AKT/mTOR pathway in liver fibrosis caused by Clonorchis sinensis in mice. Methods Six-week-old female C3H/HeN mice were randomly divided into two groups：a normal control group and an infected group（n=5for each group）.Mice in the infected group were each orally infected with 45 metacercariae.Mice in the normal control group were similarly administered an equal amount of normal saline.The liver tissues of mice were harvested to observe fibrotic changes using Masson staining 28 days post-infection.The expression and distribution of the proteinsα-SMA and p-mTOR were determined immunohistochemically.The levels ofα-SMA,COL1 and TGF-βmRNA in the liver tissue were quantified with real-time quantitative PCR（qRT-PCR）,and the levels ofα-SMA,TGF-β,p-PI3 K,p-AKT,p-mTOR protein were determined with Western blotting. Results Collagen deposition increased significantly in the fibrotic liver of infected mice than in the control group.Expression of bothα-SMA and p-mTOR increased markedly in the infected group than in the control group（t=-8.374,P〈0.01;t=-9.545,P〈0.01）.According to qRT-PCR,the expression ofα-SMA,COL1,and TGF-βmRNA was strongly up-regulated（t=-3.553,P〈0.01;t=-3.448,P〈0.01;t=-3.052,P〈0.01）in the liver of mice infected with C.sinensis.According to Western blotting,the level ofα-SMA,TGF-β,p-PI3 K,p-AKT,and p-mTOR protein was also strongly up-regulated（t=-7.076,P〈0.01;t=-5.054,P〈0.01;t=-6.941,P〈0.01;t=-8.123,P〈0.01;t=-7.445,P〈0.01）in the liver of mice infected with C.sinensis. Conclusion The PI3K/AKT/mTOR pathway may play an important role in the process of liver fibrosis induced by C.sinensis.