血管内皮生长因子基因转染骨髓间充质干细胞移植对肺气肿大鼠的疗效

Effect of Bone Marrow Mesenchymal Stem Cells Transfected With Human Vascular Endothelial Growth Factor Gene for Rats With Pulmonary Emphysema

目的:观察血管内皮生长因子(VEGF)基因转染大鼠骨髓间充质干细胞(MSCs)移植对肺气肿大鼠肺泡壁细胞的修复作用。方法:pcDNA3.1-hVEGF转染雄性Lewis大鼠MSCs。雌性Lewis大鼠随机分为4组:正常对照组、肺气肿组、单纯MSCs移植组,hVEGF移植组。观察MSCs移植28d后肺组织形态学变化,支气管肺泡灌洗液细胞分类计数,ELISA法检测VEGF含量。Y染色体荧光原位杂交(Y-FISH)示踪雄性大鼠MSCs在雌性肺气肿大鼠肺组织内的植入情况;采用TUNEL法检测肺泡壁细胞凋亡,免疫组化法检测Bcl-2蛋白的表达。结果:hVEGF移植组肺气肿改变较肺气肿组、单纯MSCs移植组减轻;支气管肺泡灌洗液炎性细胞数较后2组减少;肺组织VEGF含量较后2组增多;肺泡壁细胞凋亡指数明显低于后2组;Bcl-2染色阳性细胞百分比高于后2组;Y-FISH显示hVEGF移植组、单纯MSCs移植组的雌性大鼠肺组织中有Y染色体阳性的细胞。结论:真核表达载体pcDNA3.1-hVEGF可以成功地在MSCs中表达,pcDNA3.1-hVEGF转染MSCs移植能够减轻大鼠肺气肿样改变,其疗效优于单纯MSCs移植治疗。

Objective： To evaluate the effect of mesenchymal stem cells （MSCs） transfected with human vascular endothelial growth factor （VEGF） gene for rats with pulmonary emphysema. Methods： MSCs from male lewis rats were transfected with the pcDNA3. 1-hVEGF control vector. Female Lewis rats were randomly divided into four groups： normal control group, emphysema group, emphysema ＋ MSCs transplantation group, emphysema ＋ hVEGF-transfected MSCs transplantation group. Morphologie changes of the lung tissue were observed 28 Jays after treatment. The total cell number of bronchoalveolar lavage fluid were counted, and the content of VEGF was de tected by ELISA. The engraftment of male bone marrow MSCs in female recipient lung was determined by Y chromosome fluorescent in situ hybridization （Y-FISH）. The apoptosis of the lung cells was assessed by TUNEL stai ning. The expression of Bcl-2 were determined by immunohistochemical staining. Results： Emphysematous change in the emphysema ＋ hVEGF transfeeted MSCs transplantation group was improved compared with those in emphysema group and the emphysema ＋ MSCs transplantation group. The total cell number of bronchoalveolar lavage fluid was decreased in the emphysema ＋ hVEGF transfected MSCs transplantation group compared to the emphysema group and the emphysema ＋ MSCs transplantation group. The level of VEGF was higher in emphysema ＋ hVEGF transfected MSCs transplantation group compared with those of the emphysema group and the emphysema ＋ MSCs transplantation group. The apoptotic index of the alveolar wall cells in the emphysema ＋ hVEGF transfeeted MSCs transplantation group was less than that of the emphysema group and the emphysema ＋ MSCs transplantation group. The percentage of Bcl-2 positive cells in the emphysema ＋ hVEGF transfected MSCs transplantation group was significantly higher than that of the emphysema group and the emphysema ＋ MSCs transplantation group. Y chromo some positive cells were observed in the lungs of rats from the emphysema ＋ hVEGF transfected MSCs transplantation group and the emphysema ＋ MSCs transplantation group, Conclusions： The hVEGF gene could expressed in MSCs. Trans plantation of MSCs transfected with hVEGF gene can improve emphysematous changes than single cellular therapy.