Wnt信号通路参与实验性小鼠肝纤维化的研究  

Wnt signal pathway is involved in experimental hepatic fibrosis in mice

作　　者：毕婉蓉[1] 邢海林[2] 金彩霞[3] 

机构地区：[1]同济大学附属同济医院分院消化内科,上海200092 [2]同济大学附属同济医院普外科,上海200065 [3]同济大学医学院干细胞研究中心,上海200092

出　　处：《同济大学学报（医学版）》2014年第4期41-45,共5页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金(8107034)

摘　　要：目的探讨Wnt信号通路对实验性小鼠肝纤维化(hepatic fibrosis,HF)的促进作用。方法建立小鼠CCl4诱导HF模型,随机分为正常对照组、HF模型组与Wnt处理组。体内实验对小鼠腹腔注射Wnt3A与Wnt11各5 ng/ml,每周2次,4周后获得肝组织;体外实验Wnt处理组为正常小鼠肝细胞在含有Wnt 5 ng/ml无血清培养基培养2周后,RT-PCR法检测肝细胞α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、E-钙黏附素(E-cadherin,E-cad)、Wnt3A,Wnt5A与Wnt11。免疫荧光显微镜及激光共聚焦定位法分析肝细胞形态学变化。对培养细胞进行诱导分化,Massion染色法观察。结果体内试验HF小鼠肝细胞Wnt3A与Wnt11的免疫荧光分析及Massion染色显示,经Wnt3A与Wnt11处理后的小鼠肝上皮细胞标志(E-cad)减低,肝间质细胞标志(α-SMA)升高(P<0.05),发生了上皮-间质转化(epithelial-mesenchymal transitions,EMT)。体外试验HF小鼠肝细胞RT-PCR分析表明:HF组与Wnt处理组与正常对照组比较α-SMA、Wnt3A、Wnt5A、Wnt11表达明显增加,E-cad表达明显减少(P<0.05)。病理学支持Wnt处理组显示了类似HF的小鼠肝细胞发生EMT的形态学改变。结论研究表明Wnt信号通路参与了小鼠HF的过程,其机制可能是在实验性小鼠肝上皮细胞中通过下调内源的某些自分泌信号抑制因子,而诱导细胞启动EMT程序,从而导致HF。Objective To investigate the role of Wnt signal pathway in experimental hepatic fibrosis （HF） in mice. Methods CCl4 induced HF mice were randomly divided into model group, normal control group and Wnt group. In vivo experiments mice were injected with Wnt3A and Wntl 1 5 ng/ml respectively, 2 times a week, the hepatic tissue samples were obtained after 4 weeks. In vitro experiments the normal mouse liver ceils were cultured in serum-free medium containing Wnt 5 ng/ml for 2 weeks, then the a-smooth muscle actin （（x-SMA）, E-cadherin （E-cad）, Wnt3A, Wnt5A and Wntll were detected with semi quantitative RT-PCR. The cell morphology was observed by fluorescence microscopy and confocal laser positioning microscopy. The cultured cells were induced for differentiation and observed with Massion staining. Results Massion staining and immunofluorescence showed that epithelial cell marker E-cad decreased, mesenchymal cell marker a-SMA increased in liver cells （P 〈 0. 05 ） after Wnt3A and Wntll treatment in vivo, indicating epithelial mesenchymal transition （EMT）. RT-PCR analysis of in vitro experiments showed that expression of α-SMA, Wnt3A, Wnt5A and Wntll were significantly increased, while E-cad were decreased significantly （P 〈 0.05 ） in HF group and Wnt-treated group compared with the normal control group. Pathology examination also showed the morphological changes of EMT in mouse HF liver cells after Wnt treatment. Conclusion Results suggest that Wnt signaling pathway is involved in hepatic fibrosis in mice, which may be associated with the epithelial mesenchymal transition in liver cells.

关 键 词：肝纤维化 肝细胞 上皮-间质转换 

分 类 号：R363[医药卫生—病理学]