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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 effect of transfection of mouse CXCR4 adenovirus (Ad-mCXCR4) on the homing of damaged bone marrow mesenchymal stem cells (MSCs). Methods: Mouse MSCs were isolated and amplified in vitro. CXCR4 gene was obtained from liver of mice. The adenovirus vector Ad-mCXCR4 was constructed and identified by homologous recombination. MSCs were transfected with Ad-mCXCR4 The vector Ad-vector and untransfected MSCs were used as control. The expression of CXCR4 protein in each group was detected by Western blot. The mice were injected CCl4-induced liver injury model and then randomly divided into 3 groups. -mCXCR4, Ad-vector and untransfected MSCs 48 h after laser confocal laser scanning microscopy MSCs homing to the damaged liver tissue situation. Results: The mouse CXCR4 adenovirus vector Ad-mCXCR4 was constructed successfully. CXCR4 protein was highly expressed in MSCs transfected with Ad-mCXCR4, while CXCR4 protein was not expressed in Ad-vector and untransfected MSCs. Compared with the mice injected with Ad-mcXCR4-MSCs, the number of GFP positive cells in mice injected with Ad-mCXCR4-MSCs was significantly increased [(21.25 ± 1.56) vs. (5.42 ± 0.81)] (P <0.01). Conclusion: Gene modification can increase the expression of CXCR4 in MSCs. MSCs with high expression of CXCR4 can increase the homing to damaged liver.