Huang Jia-quan, Zhu Jun, Li Lan, Jiao Yun-tao, Xu Lei, Tao Ran, Fan Xiang-xue, Ma Ke, Guo Wei, Ning Qin
Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2012 Feb 29;30(1):20-6.
To establish the model of hepatic fibrosis in mice infected with Schistosoma japonicum and observe the expression of transforming growth factor-beta1 (TGF-beta1) and connective tissue growing factor (CTGF) in mice model.
Thirty BALB/c mice were randomly assigned to control group and model group. Each mouse of model group was infected with (30 +/- 1) S. japonicum cercariae through the abdominal skin. Serum samples were collected at 4, 6, 8, 10, and 12 weeks after infection, and were analyzed for the levels of ALT and AST. Pathological changes and proliferation of hepatic collagen fibers in liver tissue were observed after HE staining and Masson staining. Immunohistochemistry and real-time PCR were used to detect the protein and mRNA expression of CTGF and TGF-pl.
6-12 weeks after infection, there was significant difference in ALT and AST between model group and control group (P43.05). At 12th week, ALT [(173.53 +/- 31.12) U/I] and AST [(301.00 +/- 34.87) U/LI in model group were higher than those in control group [(42.00 +/- 3.53) and 96.58 +/- 11.26) U/L] . In model group, egg granulomas formed in the liver, and the formation of hepatic fibrosis was significant in portal areas, and there was tree-like hepatic fibrosis around the portal vein branch. 8 weeks after infection, hepatic fibrosis area in mice of model group increased considerably, and there was significant difference in percentage of positive area of collagen between 12th week [(23.83 +/- 1.68) %] and control group [(1.23 +/- 0.14) %] (P < 0.05). 10 and 12 weeks after infection, the percentage of positive area of TGF-beta1 [(22.34 +/- 2.58)% and (25.82 +/- 3.01) %] and CTGF [(1 l.32 +/- 2.44)% and (14.51 +/- 2.05) %] was higher respectively than that of the control [(2.56 +/- 0.87)%, and (1.09 +/- 0.73)% (P < 0.05). 6, 8, 10, and 12 weeks after infection, both TGF-beta1 and CTGF mRNA increased gradually, higher than that in control group (P < 0.05). 10 weeks after infection, TGF-beta1 mRNA relative transcription level was the highest (0.0721 +/- 0.0187) and it was 0.0089 +/- 0.0037 in control group. CTGF mRNA relative transcription level reached the highest value (0.1136 +/- 0.0365) in 12 weeks after infection, while it was 0.0293 +/- 0.0184 in control group. CTGF mRNA expression was positively correlated with the duration of infection (r = 0.927, NO.05).
The area and cell types of TGFb1 positive expression is the same as that of CTGF in liver tissue of schistosome infected mice (BALB/c). CTGF mRNA expression is significantly related to the duration of infection, but it is not the case for TGFbl.
建立日本血吸虫感染小鼠肝纤维化模型，观察该小鼠模型中转化生长因子-β1（TGF-β1）和结缔组织生长因子（CTGF）的表达。
将30只BALB/c小鼠随机分为对照组和模型组。模型组每只小鼠经腹部皮肤感染（30±1）条日本血吸虫尾蚴。感染后4、6、8、10和12周采集血清样本，分析谷丙转氨酶（ALT）和谷草转氨酶（AST）水平。经苏木精-伊红（HE）染色和Masson染色后，观察肝组织的病理变化及肝胶原纤维增生情况。采用免疫组织化学和实时荧光定量聚合酶链反应（real-time PCR）检测CTGF和TGF-β1的蛋白及mRNA表达。
感染后6-12周，模型组与对照组的ALT和AST水平有显著差异（P<0.05）。第12周时，模型组的ALT[（173.53±31.12）U/L]和AST[（301.00±34.87）U/L]高于对照组[（42.00±3.53）和（96.58±11.26）U/L]。模型组肝脏形成虫卵肉芽肿，门静脉区肝纤维化形成明显，门静脉分支周围有树状肝纤维化。感染后8周，模型组小鼠肝纤维化面积显著增加，第12周胶原阳性面积百分比[（23.83±1.68）%]与对照组[（1.23±0.14）%]相比有显著差异（P<0.05）。感染后10周和12周，TGF-β1阳性面积百分比[（22.34±2.58）%和（25.82±3.01）%]和CTGF阳性面积百分比[（11.32±2.44）%和（14.51±2.05）%]分别高于对照组[（2.56±0.87）%和（1.09±0.73）%]（P<0.05）。感染后6、8、10和12周，TGF-β1和CTGF mRNA均逐渐升高，高于对照组（P<0.05）。感染后10周，TGF-β1 mRNA相对转录水平最高（0.0721±0.0187），对照组为0.0089±0.0037。CTGF mRNA相对转录水平在感染后12周达到最高值（0.1136±0.0365），对照组为0.0293±0.0184。CTGF mRNA表达与感染时间呈正相关（r=0.927，P<0.05），而TGF-β1并非如此。
在日本血吸虫感染小鼠（BALB/c）的肝组织中，TGF-β1阳性表达的面积和细胞类型与CTGF相同。CTGF mRNA表达与感染时间显著相关，而TGF-β1则不然。
