不同数目骨髓间充质干细胞移植对大鼠肺损伤的抑制作用

目的研究不同数目的骨髓间充质干细胞（mesenchymal stem cells,MSCs）静脉移植对野百合碱（monocrotaline,MCT）诱导大鼠肺损伤的抑制作用。方法全骨髓贴壁法培养Wistar大鼠MSCs,取第3～5代细胞进行移植。健康雄性Wistar大鼠20只由颈外静脉移植MSCs,按移植细胞个数分为5×105组、1×106组、5×106组和对照（生理盐水）组（n=5）,测定移植前、移植后5 min,30 min及24 h的RVSP。另40只随机分组（n=10）：①MCT/MSCs 5×105组;②MCT/MSCs 1×106组;③MCT组;④对照组,腹腔注射60 mg/kgM CT（对照组注射等量生理盐水）,同时分别移植MSCs 5×105个、1×106个或等量PBS液体。4周后检测大鼠右心室收缩压（right ventricularsystolic pressure,RVSP）、RV/（LV＋S）重量比值;肺组织苏木素-伊红染色、地衣红染色和平滑肌Actin免疫组织化学染色。统计数据采用SPSS 11.0软件,对各组数据进行单因素方差分析检验。结果少于1×106个MSCs颈外静脉移植是安全的。MSCs颈外静脉移植4周后,MCT/MSCs 1×106组RVSP（35.6±8.4）mmHg与MCT组（47.2±10.5）mmHg相比明显下降（P〈0.05）,心室比0.3572±0.0923明显低于MCT组0.4454±0.0935（P〈0.05）,而MCT/MSCs 5×105组RVSP为（42.5±11.3）mmHg,心室比0.4003±0.0725,与MCT组相比无明显下降（两者P〉0.05）;病理染色可见肺组织肺小动脉中,MCT/MSCs 1×106组中膜厚度分布为（19.2±3.8）%,较MCT组（26.4±4.9）%明显变薄（P〈0.05）;而MCT/MSCs 5×105组（23.3±3.6）%较MCT组相比无显著差异（P〉0.05）。结论 MSCs颈外静脉移植对MCT诱导的肺损伤具有抑制作用,1×106个细胞较5×105个细胞移植抑制作用显著。

Preventive Effects of Different Number of Mesenchymal Stem Cells Transplantation on Lung Injury in Rats

Objective To observe effects on lung injury after different number of mesenchymal stem cells transplantation in rats.Methods MSCs of Wistar rats were isolated and cultivated by bone marrow adherent culture.The cells between the third and fifth passages were prepared for transplantation.20 male Wistar rats were divided into four groups by the number of MSCs transplanted intravenously： the 5×105 group、the 1×106 one、the 5×106 one and the control one（n=5）.RVSPs were measured at the time of pro-transplantation,5 min,30 min and 24 h post-transplantation.The other 40 rats were randomly divided into four groups： ①the MCT/MSCs 5×105 group;②the MCT/MSCs 1×106 one;③the MCT one;④the control one.Rats were injected intraperitoneally with 60 mg·kg1MCT,except the control group with physiological saline.Phosphate buffered saline with 5×105 MSCs and 1×106 MSCs were transplanted into rats through external jugular vein in the first two groups respectively.At the forth week after cell administration,RVSPs were measured and lungs＇ tissues were stained with Hematoxylin-eosin staining,lichen dyed red staining and smooth muscle Actin immunohistochemistry.Lungs＇ pathological changes were observed.All data were analyzed statistically by one way analysis of variance using SPSS 11.0.Results It was safe to transplant MSCs less than 1×106 intravenously.At the forth week after MSCs administration,RVSP and ventricular ratio significantly decreased in the MCT/MSCs 1×106 group compared to that in MCT group（47.2±10.5 mmHg vs 35.8±8.4 mmHg,0.3572±0.0923 vs 0.4454±0.0935,respectively,both P 0.05）,but they didn＇t differ statistically in MCT/MSCs 5×105 group compared to in the MCT one.By pathological staining,medium thickness of the pulmonary arterioles was observed comparatively thinner in the MCT/MSCs 1×106 group than those in the MCT group.In contrast they had no significant difference between in the MCT/MSCs 5×105 group and in the MCT one（P 0.05）.Conclusion Intravenous MSCs administration can prevent lungs injury induced by monocrotaline and 1×106 MSCs administration has better effects than 5×105 MSCs＇.