静脉注射大鼠骨髓MSCs缓解新生大鼠高氧肺损伤
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广东省政府重大攻关课题(B30502)


Effects of marrow-derived mesenchymal stem cells on the lung damage in new born rats suffering from hyperoxia
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    摘要:

    目的:研究大鼠骨髓间充质干细胞(mesenchymal stem cells, MSCs)对高氧新生大鼠肺组织血管内皮生长因子(Vascular endothelial cell growth factor,VEGF)表达的影响。方法:采用贴壁选择法分离、培养、扩增大鼠骨髓来源MSC,并以5-溴脱氧尿嘧啶核苷(5-bromo-2-deoxyuridine,BrdU)进行标记;3日龄清洁级SD新生大鼠32只,随机分为高氧组(A组)、高氧干预组(B组)、正常干预组(C组)、对照组(D组)4组;A、B二组均置大鼠于95%氧环境下7天后,B组每只动物尾静脉注射5 × 104的MSCs,A组每只注射磷酸盐缓冲液(PBS)50 μl;而C、D二组则置于正常空气环境下,同时分别给予每只大鼠尾静脉注射含5 × 104MSC及PBS 50 μl。注射后72 h(13日龄)处死全部动物,肺组织病理学检测辐射状肺泡计数(radical alveolar counts,RAC),免疫组织化学检测VEGF 吸光度(A)值、BrdU表达情况。结果:仅二个注射MSC组(B、C组)可见BrdU阳性表达,高氧组RAC值及均显著低于对照组,而给予MSC均可使其明显上升,近于对照组。结论:给新生大鼠静脉注射MSC对高氧肺损伤的保护作用可能与增加肺组织VEGF表达有关。

    Abstract:

    Objective:To investigate the effects of marrow-derived MSCs on the lung damage in new born rats suffering from hyperoxia. Methods:Marrow-derived MSCs were separated, cultured, amplificated and labeled with BrdU. Thirty-two three-day-old SD rats were randomly divided into four groups(A,B,C,and D). Rats in group B and group A were exposed to 95% oxygen for 7 days, then they were injected with MSCs of 5 × 104 and PBS into tail vein respectively. Rats in group C and group D lived in normal air and were injected with MSCs of 5 × 104 and PBS into tail vein respectively at the same time. Immunohistochemistry(IHC) staining was used to determine the expression of 5-bromo-2-deoxyuridine(BrdU) and vascular endothelial cell growth factor(VEGF) 72 h later. Radical alveolar counts(RAC) of the lungs in the rats were counted under light microscope. Results:The labeled MSCs were detected in the lungs of rats treated with MSCs, but no BrdU-positive cells were found in two untreated groups(A and D). There were significant differences in the levels of VEGF and RAC among four groups(P < 0.05, P < 0.01 respectively). Conclusion:BrdU-labeled marrow-derived MSCs can be implanted into lungs of new born rats suffering from hyperoxia. The protective effect of MSCS on the damaged lungs induced by hyperoxiab might be related to the increased expression of VEGF.

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田兆方,杜 江,王 斌,洪小杨,封志纯.静脉注射大鼠骨髓MSCs缓解新生大鼠高氧肺损伤[J].南京医科大学学报(自然科学版),2007,(3):239-241248

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  • 收稿日期:2006-12-01
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