T淋巴细胞介导的小鼠免疫性肝纤维化模型的建立及形态学观察

Establishment and morphological study of a novel T cell mediated immune hepatic fibrosis model in BALB/C mice

目的 建立刀豆蛋白A诱导的T淋巴细胞介导的小鼠免疫性肝纤维化模型并进行形态学观察。方法 BALB/C小鼠随机分组。模型组、CD4抗体处理组、CD8抗体处理组、地塞米松处理组和裸鼠组静脉注射刀豆蛋白A ;CD4、CD8抗体处理组、地塞米松注射组和裸鼠组共注射 5周 ,模型组共注射 2 0周 ,分别于相应的时间点取血、取肝脏组织统一保存。检测ALT水平 ,进行肝脏组织苏木精 伊红染色、免疫组织化学染色和VanGieson胶原纤维染色。结果 模型组接受刀豆蛋白A刺激后ALT水平明显高于正常对照组 ,而CD4抗体处理组、地塞米松处理组和裸鼠组ALT水平明显低于模型组 ,CD8抗体处理组ALT水平则与阳性组无明显差异。肝组织冰冻切片免疫组织化学染色提示肝组织内淋巴细胞浸润以CD4+ T淋巴细胞为主。模型组病理检查提示肝纤维化发生。

Objective To establish and to study the morphology of a novel T cell mediated immune hepatic fibrosis model induced by repeated injections of ConcanavalinA in BALB/C mice. Methods BALB/C mice were divided into various groups; normal control group treated with saline, model group, pretreated with monoclonal anti mouse CD4 and monoclonal anti mouse CD8 groups, dexamethasone pretreated group and weekly Concanavalin A administered nude mice group. In the nonoclonal anti mouse CD4, monoclonal anti mouse CD8, dexamethasone and the nude mice groups, sera and liver samples were collected at different peroids. At 1,2,3,4,5,12 and 20 weeks after concanavalin A administration and the sera were stored under -30°C until they were measured. The livers of different groups were fixed in 10% formalin for routine light microscopy or frozen in optimal cutting temperature (O.C.T.) in liquid nitrogen for immunohistochemical study of CD4 + or CD8 + T cells. Fibrosis was assessed with Van Gieson staining and the fibrous tissue was measured with MPIAS500 imaging system.Results The levels of ALT in the model group were higher than those in the normal group. The levels of anti mouse CD4 pretreated group, dexamethasone pretreated group and nude mice group. The levels of ALT in anti mouse CD8 pretreated group were similar to those in the model group. Immunohistochemical staining for CD4 + or CD8 + T cells indicated that the infiltrating lymphocytes in liver parenchyma were mainly CD4 + T lymphocytes. The change in the HE and Van Gieson stain in the model group mice indicated presence of fibrogenesis. Conclusion Repeated administration of Concanavalin A can induce T cell mediated immune hepatic fibrosis model in BALB/C mice.