分泌型卷曲相关蛋白重组腺病毒载体对小鼠肝纤维化的抑制作用  

Transfection with recombinant adenovirus vector expressing secreted frizzled related protein inhibits liver fibrosis in mice

作　　者：毕婉蓉[1] 杨长青[2] 邢海林[3] 

机构地区：[1]同济大学附属同济医院分院消化科,上海市200092 [2]同济大学附属同济医院消化内科,上海市200065 [3]同济大学附属同济医院普外科,上海市200065

出　　处：《世界华人消化杂志》2014年第29期4379-4385,共7页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.8107034;上海市同济医院横向基金资助项目;No.HX1401~~

摘　　要：目的:成功构建携带分泌型卷曲相关蛋白(secreted frizzled related protein,SFRP)基因的腺病毒载体,并探讨SFRP对实验性小鼠肝纤维化(hepatic fibrosis,HF)的抑制作用.方法:用脂质体转染法将重组腺病毒pAdTrack-PPARγ2-CMV和SFRP5基因共转染CCl4诱导小鼠HF模型肝细胞,成功构建的重组腺病毒Ad-SFRP5感染HF细胞.采用Western blot、免疫荧光显微镜及激光共聚焦定位分析观察Ad-SFRP5抑制HF作用.结果:感染的重组腺病毒载体在小鼠HF细胞中有较高的SFRP5蛋白的稳定表达.免疫双荧光显示Ad-SFRP5抑制HF小鼠肝细胞α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和成纤维细胞特异蛋白-1(fibroblastspecific protein-1,FSP-1)的表达.病理检测支持Ad-SFRP5抑制HF小鼠肝细胞上皮-间质转换(epithelial-mesenchymal transitions,EMT)的发生(SFRP5处理组与正常对照组相比,P<0.05).结论:我们成功构建含有小鼠SFRP5基因的重组腺病毒,他可有效提高小鼠HF细胞中SFRP5基因的表达水平,并可抑制EMT程序,从而抑制HF.AIM： To construct a recombinant adenovirus vector expressing secreted frizzled related protein（SFRP）, and to assess the inhibitory effect of SFRP overexpression on experimental hepatic fibrosis（HF） in mice.METHODS： Liposome-mediated transfection was used to introduce recombinant adenovirus pAd-Track-PPARγ2-CMV carrying the SFRP5 gene into liver cells from mice with CCl4 induced HF to obtain recombinant adenovirus infected HF cells. Western blot, immunofluorescence microscopy and confocal laser microscopy were used to analyze the inhibitory effect of SFRP overexpression on experimental HF. RESULTS： Cells infected with the recombinant adenovirus vector had higher expression of SFRP5 protein. Double immunofluorescence showed SFRP5 over expression inhibited the expression of α-smooth muscle actin（α-SMA） and fibroblast-specific protein-1（FSP-1） in liver cells. Pathological examination revealed that SFRP5 over expression significantly reduced the occurrence of epithelial-mesenchymal transition（EMT） in liver cells（SFRP group vs control group, P〈0.05）. CONCLUSION： We have successfully constructed a recombinant adenovirus containing the SFRP5 gene, and it can effectively increase the expression of SFRP5 gene in mouse HF cells, thereby inhibiting the occurrence of EMT and HF.

关 键 词：腺病毒 肝纤维化 分泌型卷曲相关蛋白 

分 类 号：R575[医药卫生—消化系统]