骨髓间充质干细胞应用于急性肺损伤修复的实验研究

Investigation of treatment by BMSCs for Acute Lung Injury

目的探讨应用骨髓间充质干细胞(BMSCs)治疗严重肺部疾病的可能性,为临床上利用干细胞方法治疗严重肺部疾病提供参考。方法选用6周龄健康Wistar大鼠(约150克),分离和培养BMSCs。取2代BMSCs,免疫荧光法检测细胞CD29、CD44、CD34、CD45表面标志。20只大鼠分为两组,一组经尾静脉注入脂多糖,另一组经尾静脉注入脂多糖和经溴脱氧尿嘧啶标记的骨髓间充质干细胞。另有2只大鼠:1只为正常对照,1只为干细胞对照。观察比较大鼠呼吸状态,呼吸频率,活动状态,摄食情况,口鼻分泌物情况;留取肺组织标本行病理组织学检查,并进行分析比较。结果原代培养的BMSCs一般9-10天即可长到80-90%融合,第2代后细胞的形态比较均一;免疫荧光法检测BMSCs,CD29、CD44呈阳性反应,CD34、CD45呈阴性反应;正常对照和干细胞对照大鼠呼吸状态及活动性良好;大体观察显示,A、B两组大鼠肺脏均出现不同程度淤血水肿,但A组程度更重;肺脏病理切片显示,A组大鼠肺组织出现了明显的水肿,充血,甚至肺实变的情况;B组大鼠肺组织损伤程度较轻,且没有肺实变情况发生。结论经外源性BMSCs治疗干预的急性肺损伤大鼠,肺脏病理改变与对照组相比有所减轻;外源性BMSCs在急性肺损伤的大鼠肺脏中可归巢定植。

Objective To investigate the possibility of using Bone Marrow Stem Cells （BMSCs） to treat severe diseases of the lung ,and provide the reference of treatment for severe diseases of the lung . Methods The BMSCs of Wistar rats of six weeks （150g） were isolated and cultured .We used the second passage cells to detect cell surface markers by the immunofluorescence assay .Twenty rats were divided in-to two groups ：group A injected with only LPS ,and group B injected with both LPS and BMSCs marked with Bromodeoxyuridine （BrdU ） .The respiratory condition ,respiratory frequency ,activity ,eating and secretion in mouth and nose were observed .The lungs of the rats were dissected and examined pathologi-cally .The results were analyzed and compared .Results The primary cultured BMSCs get 80 - 90% con-fluence within 9 - 10days .The shape of the offspring cells had homogeneity .The cell surface markers CD29 and CD44 were positive ,whereas CD34 and CD45 were negative through the immunofluorescence as-say .The respiratory conditions and activities of the control rats were good .Some rats of both groups had dyspnea ,hyperpnea ,hypoactivity ,fasting and secretion in mouth and nose .However ,the number of suf-fered rats in A group is more .The lungs of both groups showed congestion and edema .However ,the lungs of group A were more severe . The pathological slices of the rats injected with LPS （group A ） showed edema ,congestion and consolidation of lungs ,whereas those of the rats injected with LPS and BMSCs （group B） showed mild edema and congestion of lungs .Conclusion The lung injury of the rats in-jected with both LPS and BMSCs is milder than that with LPS only .The BMSCs injected into the rats with acute lung injury can distribute in the lungs of the rats .