不同时期血吸虫肝损伤模型小鼠肝脏组织病理学及肝功能的动态观察

Dynamic Observation on Liver Pathology and Function of Mice Infected with Schistosoma Japonicum

目的观察不同时期血吸虫肝损伤模型小鼠肝脏组织病理学和肝功能的变化,为血吸虫肝损伤的研究提供理论依据。方法雄性Balb/c小鼠通过腹部皮肤贴敷尾蚴的方法制备血吸虫肝损伤模型。分别在制模后第1、3、6、9周各处死模型鼠10只,第11周和正常对照组各处死6只。取肝脏、脾脏和肾脏组织并称重,计算各个时期小鼠的肝脏指数、脾脏指数及肾脏指数;肝脏组织分别行苏木精-伊红(HE)染色和Masson染色观察肝损伤的病理变化和胶原纤维的沉积。摘除眼球取全血用全自动生化仪检测血清中的谷丙转氨酶(ALT)和谷草转氨酶(AST);放射性免疫法检测血清中的透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)和Ⅳ型胶原(Ⅳ-C)。结果血吸虫肝损伤模型制备成功。随着时间的推进,模型鼠的肝脏组织在第6周逐渐出现嗜酸性肉芽肿结节、胶原纤维及虫卵的沉积;肝脏指数和脾脏指数在第6周逐渐升高,第6、9、11周时与正常组比较差异具有统计学意义(P<0.05或P<0.01),肾脏指数无明显变化。ALT、AST在第6周达到高峰,而后下降,但均高于正常组水平(均P<0.01)。HA和PCⅢ分别在第6周和第3周达到高峰,而后下降,但仍高于正常组水平(均P<0.01);LN在第1、3、9周明显升高(均P<0.01);Ⅳ-C在第1周和第3周明显升高(均P<0.01),而后降至正常水平。ALT、AST水平与HA水平呈正相关(r=0.32、0.29,P<0.05)。结论随着时间的推进,血吸虫肝损伤模型小鼠的肝纤维化程度逐渐加重,肝脏和脾脏是晚期血吸虫病主要的损伤器官,ALT和AST可基本反映肝脏的损伤情况,HA和PCⅢ可较敏感地反映血吸虫肝损伤模型小鼠的肝纤维化程度。

Objective To observe the dynamic changes of liver pathology and function in mice infected with Schistosoma japonicum and supply evidence for the study on Schistosoma japonicum-induced liver damage.Methods Male Balb/c mice were infected subcutaneously with Schistosoma japonicum cercaria to prepare the liver damage models.Ten mice were sacrificed respectively at 1st,3rd,6th,and 9th week after infection,and 6 mice at 11th week post-infection and the normal control group.The liver,spleen and kidney were respectively taken to calculate the indexes.The liver tissues were subjected to hematoxylin-eosin（HE）and Masson staining,and the pathological changes and collagen deposition in the liver damage were observed.Aspartate transaminase（AST）and alanine transaminase（ALT）in serum were determined by using autobiochemical machine.Hyaluronic acid（HA）,laminin（LN）,precollagen Ⅲ（PCⅢ）and collagen Ⅳ（Ⅳ-C）in serum were tested by using radioimmunoassay.Results The Schistosoma japonicum-induced liver damage models were successfully established.With time over,the eosinophilic granuloma appeared,and the collagen fiber and eggs started to deposit in the liver tissue at the 6th week post-infection.As compared with normal control group,the liver and spleen indexes in the model group were increased at 6th week after infection（P0.05）,but the kidney index had no significant change.ALT and AST levels reached the peak at 6th week post-infection,and declined at 9th and 11th week,but still evidently higher than the normal levels（P0.01）.HA and PCⅢ levels respectively reached the peak at 6th and 3rd week post-infection,and decreased afterward,but obviously higher than the normal levels（P0.01）.LN level was increased to the peak at 1st,3rd and 9th week post-infection（P0.01）.Ⅳ-C level reached the peak at 1st and 3rd week post-infection,and declined to the normal level afterward.ALT and AST level had a positive correlation with that of HA（r=0.32,0.29,P0.05）.Conclusion With time over,the degree of liver fibrosis in model mice gradually aggravates.The liver and spleen are the chief organs in the terminal schistosoma disease.ALT and AST can basically reflect the damage degree of the liver,and HA and PCⅢ are comparatively sensitive to reflect the liver fibrosis degree of schistosoma liver damage model in mice.