骨髓间充质干细胞移植在肺气肿大鼠模型肺组织内定植研究

Study on the Bone Marrow Mesenchymal Stem Cell Transplantation in Rats Lung Tissue with Emphysema

目的:观察骨髓间充质干细胞移植在肺气肿大鼠模型肺组织内的定植情况。方法:选择健康SD大鼠34只,按随机数字表法分为MSCs干预组(A组,10只,慢阻肺大鼠,尾静脉输注MSC 1×106个/mL),肺气肿模型组(B组,10只,慢阻肺大鼠,尾静脉输注等体积PBS)及MSC对照组(C组,10只,正常大鼠,尾静脉输注MSCs 1×106个/mL),正常对照组(D组,4只,正常大鼠,尾静脉输注等体积PBS),采用烟熏法复制大鼠肺气肿模型。全骨髓培养法体外培养扩增雄性SD大鼠来源的MSCs,经GFP标记细胞后将其经尾静脉注入肺气肿模型SD大鼠体内,24 h内处死大鼠,取肺组织迅速冰冻切片,共聚焦激光显微镜下观察观察转染GPF的间充质干细胞在大鼠肺内定植情况。结果:成功培养具有分化潜能的骨髓间充质干细胞,MSCs传至第4代时有99.5%表达CD44、99.6%表达CD29等间充质干细胞表面标志,仅有0.4%表达CD34、1.0%表达CD45单核细胞以及造血干细胞表型;成功复制大鼠肺气肿模型,香烟烟雾暴露组(A、B组)平均肺泡间隔为(119.0±26.2)μm,高于对照组(C、D组)的(89.8±17.3)μm,差异有统计学意义(P<0.05);平均肺泡数为(173.9±68.3)个/mm2低于对照组的(280.3±104.0)个/mm2,差异有统计学意义(P<0.05);显微共聚焦发现MSCs经尾静脉注入大鼠体内24 h后可见A组大鼠肺组织内转染绿色荧光蛋白质粒的MSCs,而B组、C组及D组均未见转染荧光。结论:骨髓间充质干细胞经尾静脉输注入后可在肺气肿模型大鼠肺内定植,为MSCs治疗慢阻肺可能提供理论依据。

Objective：To observe the situation of bone marrow mesenchymal stem cell transplantation in rats lung tissue with emphysema.Method：34 healthy rats were randomly divided into the MSCs intervention group（the group A,10 COPD rats,tail intravenous MSC 1×106cells/mL）,the emphysema model group（the group B,10 COPD rats, tail intravenous the same volume PBS）,the MSC control group（the group C,10 normal rats,tail intravenous MSCs 1×106cells/mL）and the normal control group（the group D,10 normal rats,tail intravenous the same volume PBS）. Copy the rats emphysema model used smoke,proliferated the male SD rats MSCs by whole bone marrow culture method in vitro. The cells with GFP labeled were injected into SD emphysema rats serum,killed the rats within 24 h,got the lung tissue to rapid frozen section. The homing capacity of mesenchymal stem cells with GPF transfection in rat lung was analyzed by observing pathological section.Result：The differentiation potential of bone marrow mesenchymal stem cells were successfully developed. 99.5%marrow mesenchymal stem cells expressed CD44,99.6%expressed CD29,0.4%expressed CD34 and 1.0%expressed CD45 when MSCs to the fourth generation. The model of rat emphysema was made successfully. The average alveolar interval in the cigarette smoke exposure group（the group A,B）was（119.0±26.2）μm higher than the control group（the group C,D）,and the average number of alveolar was（173.9±68.3）/mm2 lower than the control group. The MSCs with transfection in the rats after 24 h green fluorescent protein were observed in the group A,but not in the group B,C and D. Conclusion：The bone marrow mesenchymal stem cells can colonize in pulmonary emphysema model rats by tail intravenous injection. This may provide theoretical basis for MSCs in the treatment of COPD.