目的建立免疫性肝损伤模型,探讨Tim-3及Th17细胞在免疫性肝损伤发病过程中的动态变化及意义。方法 60只Wistar雄性大鼠随机分为急性组(10只)、中间组(10只)、慢性组(30只)和对照组(10只)。用刀豆蛋白A建模,在第0、2、4、6、8周时对慢性组大鼠实施体外心间取血1.5 ml,分离血清后于-20℃备用。8周建模成功后取完整肝脏并计算肝脏指数;生化分析法测血清中ALT、AST、TP及ALB的动态变化;ELISA法测血清中细胞因子Tim-3、IL-17、IL-6及IL-23的动态表达;HE染色观察肝脏的病理变化;免疫组化法检测肝脏中Tim-3和IL-17蛋白的表达。结果与对照组相比,急性组、中间组及慢性组的ALT及AST均显著升高(P〈0.05),ALB显著下降(P〈0.05)。中间组HE染色偶见假小叶,而慢性组发现有明显的炎性细胞和肝脏损伤,镜下假小叶较多见。在慢性组的不同阶段中,IL-17的动态表达是先升高,4周后开始降低(P〈0.05);Tim-3的动态表达则是先降低,4周以后开始升高(P〈0.05)。结论 Tim-3及Th17细胞参与了免疫性肝损伤的发病机制,其表达水平的变化可以在一定程度上反应病情的严重程度。
To establish immunological hepatic injury rat model and investigate the dynamic changes of Tim-3and Th17 cells in these process and its significance, Sixty male Wistar rats were recruited and randomly divided into acute group(n=10), middle group(n=10), chronic group(n=30) and control group(n=10). The immunological hepatic injury model was built with Con A and 1.5 ml blood was gathered by apex cordis from chronic group at the zero, the second, the fourth, the sixth and the eighth week, then the serum was separated and stored at-20 ℃. Eight weeks after, whole liver was resected and the liver index was calculated. The biochemical analysis method was applied to determine dynamic change of ALT, AST, TP and ALB in serum, while enzyme-linked immunosorbent assay(ELISA) was used detect the dynamic expressions of Tim-3, IL-17, IL-6 and IL-23. HE staining was conducted in liver tissue to observe the pathological variations; immunohistochemical staining was used to test the expression of Tim-3 and IL-17 proteins in livers. Data showed that compared with the control group, the levels of ALT and AST in other groups were increased significantly(P 〈 0.05), while ALB was decreased remarkably(P 〈 0.05). Inflammatory cells,hepatic injure and false lobules were found in the liver of chronic group with HE staining, while only few false lobules were observed in the middle group. In the different stages of chronic group, the expression of IL-17 was risen at first,but fallen 4 weeks after(P 〈 0.05), while the expression of Tim-3 was decreased at first then increased from the fifth week(P 〈 0.05). All results indicated that Tim-3 and Th17 cells may play a part in the process of immunological hepatic injury. To some extent, the changed expression levels can reflect the severity of the hepatic disease.