小鼠CXCR4腺病毒的构建及其转染对MSCs向受损肝脏归巢的影响  

Adenovirus carrying mouse CXCR4 gene and its effect on MSCs homing to injured liver

作　　者：冯欢[1] 王峥[1] 梁平[1] 李靖[1] 王威巍[1] 李洪艳[1] 黄小兵[1] 

机构地区：[1]第三军医大学附属新桥医院肝胆外科,重庆400037

出　　处：《中国普通外科杂志》2012年第7期811-815,共5页China Journal of General Surgery

基　　金：重庆市自然科学基金资助项目(CSTC;2008BB5118)

摘　　要：目的:观察小鼠CXCR4腺病毒(Ad-mCXCR4)转染对骨髓间充质干细胞(MSCs)受损肝脏归巢的影响。方法:分离小鼠MSCs并体外扩增、鉴定;从小鼠肝脏组织中获得CXCR4目的基因,用同源重组法构建腺病毒载体Ad-mCXCR4并鉴定;用Ad-mCXCR4转染MSCs,以转染空载体Ad-vector和未转染的MSCs为对照,然后用Western blot法检测各组细胞CXCR4蛋白的表达;小鼠注射CCl4诱导肝损伤模型后随机分为3组,分别通过尾静脉注射转染Ad-mCXCR4,Ad-vector和未转染的MSCs,48 h后用激光共聚焦观察各组MSCs归巢到受损肝组织的情况。结果:成功构建小鼠CXCR4腺病毒载体Ad-mCXCR4;MSCs转染Ad-mCXCR4后CXCR4蛋白呈高表达,而转染Ad-vector和未转染的MSCs无CXCR4蛋白表达;注射未转染MSCs组小鼠肝脏未见绿色荧光蛋白阳性细胞,注射Ad-mCXCR4-MSCs组小鼠较注射转染Ad-vector-MSCs组小鼠肝脏绿色荧光蛋白阳性细胞明显增加[(21.25±1.56)vs.(5.42±0.81)](P<0.01)。结论:基因修饰可提高MSCs的CXCR4表达,高表达CXCR4的MSCs向受损肝脏归巢增加。Objective： To observe the homing of mesenchymal stem cells（MSCs） transfected with adenovirus containing mouse CXCR4 gene to the injured liver. Methods： The mouse MSCs were isolated,expanded in vitro and then identified.The target CXCR4 gene was obtained from mouse liver,and the adenovirus vector Ad-mCXCR4 was constructed by homologous recombination and identified.MSCs were transfected with Ad-mCXCR4,with the MSCs transfected with empty vector Ad-vector or without transfection serving as controls,and the CXCR4 protein expression in each cell group was determined by Western blot analysis.Mice were randomly divided into three groups after the establishment of CCl4-induced liver damage,and then,the Ad-mCXCR4 transfected MSCs,Ad-vector transfected MSCs or non-transfected MSCs were injected into the mice through tail vein,respectively.At 48 h after injection,the MSCs homing signals from the injured liver tissues of each group were detected by laser confocal microscopy. Results： Recombinant adenovirus vector Ad-mCXCR4 was constructed successfully.The MSCs transfected with Ad-mCXCR4 presented a strong CXCR4 expression,while the MSCs transfected with Ad-vector or without transfection had no CXCR4 expression.No green fluorescent protein-positive cells were noted in the liver of mice injected with non-transfected MSCs,and the number of green fluorescent protein-positive cells in the liver of mice injected with Ad-mCXCR4-MSCs was significantly higher than that of mice injected with Ad-vector-MSCs [（21.25±1.56） vs.（5.42±0.81）]（P0.01）. Conclusion： CXCR4 expression can be elevated in MSCs by gene modification technique and the homing of CXCR4-overexpressing MSCs to the injured liver is enhanced.

关 键 词：间质干细胞移植 归巢 受体 CXCR4 肝 

分 类 号：R329.29[医药卫生—人体解剖和组织胚胎学]