血吸虫病肝纤维化小鼠中肝星状细胞迁移功能改变的实验研究

目的探讨影响日本血吸虫病肝纤维化小鼠肝组织中肝星状细胞(HSCs)迁移运动功能变化的相关因素。方法 SPF级6⁸周龄Balb/c小鼠16只,随机分为模型组(8只)和对照组(8只),以血吸虫尾蚴腹部贴附法建立感染模型,正常组予以生理盐水代替。于感染后8周末处理小鼠,取部分肝组织石蜡包埋,进行病理学评估,免疫荧光染色检测HSCs(α-SMA,红光)运动蛋白Fascin(绿光)的表达;另取部分肝组织,采用Real-time PCR方法检测迁移诱导因子转化生长因子β1(TGF-β1)、血小板源性生长因子(PDGF)以及单核细胞趋化因子1(MCP-1)的表达以及HSCs运动蛋白α-SMA、Fascin的表达。结果 8周末时,模型组小鼠肝组织中已形成明显肝纤维化。模型组小鼠肝组织中TGF-β1、PDGF以及MCP-1的基因表达水平分别是对照组的30倍、14倍及14倍,差异具有统计学意义(P=0.033、P=0.039以及P=0.037);同时,模型组中HSCs运动相关蛋白α-SMA和Fascin的基因表达水平分别是对照组的9倍和5倍,差异具有统计学意义(P=0.004、P=0.018);荧光共聚焦结果提示,模型组小鼠肝组织中α-SMA(红色)和Fascin(绿色)表达部位一致,集中在虫卵周围肝纤维化区域,较对照组二者表达明显增加,且红绿光分布多重叠。结论诱导HSCs运动迁移的因子表达增加和HSCs自身的运动相关蛋白表达增加均有利于HSCs运动迁移能力增强。

Alterations of hepatic stellate cells on movement abilities in mice infected with Schistosoma japonicum

Objective To analyze the relevant changes of hepatic stellate cells （HSCs） migration in mice with Schistosoma japonicum infection. Methods A total of 16 SPF balb/c mice aged 6-8 weeks, were randomly divided into two groups, namely, control group （n=8） and infected group （n=8）. The mice from the infected group were suffered from skin infection by schistosome cercariae, while the mice in control group were given an equal volume of saline instead. The mice were sacrificed at the end of the eighth week. Liver lesions were fixed immediately in 10% neutral-buffered formalin and processed into paraffin sections. Pathological changes and proliferation of hepatic collagen fibers in liver tissue were observed after HE staining and Masson staining. Changes in Fascin expression （green） of HSCs （red） were visualized by fluorescence staining and fluorescence images were recorded using confocal microscopy. By Real-time PCR, the expression of transforming growth factor-β1 （TGF-β1）, platelet-derived growth factor （PDGF）, monocyte chemotactic protein-1 （MCP-1）, α-smooth muscle actin （α-SMA） and Fascin was analyzed. Results Visible liver fibrosis was observed in the infected group at the end of the eighth week. The mRNA levels of TGF-β1, PDGF, MCP-1,α-SMA and Fascin were more than 30-fold, 14-fold, 14-fold, 9-fold, and 5-fold higher in the infected group than in control group, respectively. The differences between＆amp;nbsp;two groups were statistically significant （P=0.033, P=0.039, P=0.037, P=0.004 and P=0.018, respectively）. In addition, by confocal microscopy, Fascin immunoreactivity was almost undetectable in control group, but it was increased in the infected group along sinusoids and fibrosis area, overlapping with α-SMA, a maker of activated HSCs. Conclusion Enhancing the migration ability of activated HSCs which is a result of increasing expression of mobility-inducible factors and motility related proteins may play an important role in the progression of liver fibrosis infec