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Ang-1基因修饰的骨髓间质干细胞移植治疗脑梗死的实验研究 被引量:11

Therapeutic efficacy of Ang-1 gene-modified MSCs in cerebral infarction
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摘要 目的:观察血管生成素-1(Ang-1)和骨髓间质干细胞(MSCs)对脑梗死的联合治疗作用。方法:构建带有大鼠Ang-1基因的慢病毒载体,感染大鼠骨髓间质干细胞(rMSCs)获得可表达具有生物活性Ang-1蛋白的rMSCs(Ang-rMSCs)。制备大鼠大脑中动脉栓塞模型,随机分为对照组、Ang-rMSCs组及空载体-rMSCs(pNL-rMSCs)组。脑栓塞术后24h进行细胞移植。各组大鼠分别于术后24h、移植后1周、1月及3月用mNSS量表评价神经功能状况,移植后1周应用伊文思蓝(EB)检测血脑屏障通透性,应用荧光激发及荧光免疫组化观察移植细胞在脑内的存活、分化情况,并经组织学观察有无成瘤迹象。结果:与对照组相比,移植后1周、1月,pNL-rMSCs及Ang-rMSCs组神经功能改善明显(P<0.01),Ang-rMSCs组明显优于pNL-rMSCs组(P<0.001)。移植后3月3组间的mNSS评分差异无显著(P>0.05)。EB检测示血脑屏障通透性在pNL-rMSCs及Ang-rMSCs组低于对照组(P<0.01),Ang-rMSCs组低于pNL-rMSCs组(P<0.01)。移植后1周、1月及3月,Ang-rMSCs和pNL-rMSCs组在荧光激发下均见移植细胞在梗死灶及其周边区聚集并存活。荧光免疫组化检测示Ang-rMSCs组中移植细胞大量表达Ang-1,两移植组中均有部分细胞可同时表达神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF)或星形胶质细胞标志物神经胶质纤维酸性蛋白(GFAP)。未观察到移植细胞异常增殖等成瘤迹象。结论:Ang-1基因修饰的rMSCs移植后可迁移至脑梗死灶周围,分化表达神经细胞标志物并较长期存活。MSCs移植与其分泌的Ang-1蛋白可协同促进脑梗死后神经功能恢复。 AIM: To observe the therapeutic efficacy of Ang - 1 gene - modified mesenchymal stem cells (MSCs) in cerebral infarction. METHODS : The constructed lentiviral vector carrying the Ang - 1 gene was used to infect the rat mesenchymal stem cells (rMSCs) to establish the Ang - 1 gene - modified rMSCs (Ang - rMSCs). Adult male F344 rats were subjected to transient (2 h) middle cerebral artery occlusion (MCAO) with modified Zea Longa method.Phosphate buffered saline ( PBS, 1 mL 0. 1 mol/L, for control group ), or Ang - rMSCs suspension ( 1 mL, for Ang - rMSCs group), or rMSCs suspension ( 1 mL, for pNL - rMSCs group), were infused into tail vein of rat respectively at 24 h after MCAO ( n = 8 in each group). Functional recovery measurements using the modified neurological severity scores (mNSS) were performed at 24 h post - MCAO and 1 week, 1 month and 3 months post - transplantation, respectively. The quantitative evaluation of blood - brain barrier permeability was performed at 1 week post - transplantation. The distribution, differentiation and malignant sign of grafted rMSCs were observed with immunofluorescence staining and histological method. RESULTS : Significant neurological function improvement was observed in groups treated with Ang - rMSCs or pNL - rMSCs at 1 week, 1 month post - transplantation compared with that in control group, as evidenced by mNSS (P 〈0. 01 ). Better neurological function improvement was also found in Ang -rMSCs group than that in pNL- rMSCs group ( P 〈 0. 01 ). The results of quantitative evaluation of blood - brain barrier permeability showed that the permeability in Ang - rMSCs group was the lowest compared to those in other two groups ( P 〈 0. 01 ), and in the pNL - rMSCs group was the lower than that in control group ( P 〈 0. 01 ). The grafted rMSCs survived in Ang - rMSCs and pNL - rMSCs groups, most were localized around the ischemic focus, and a few of them expressed NSE, NF and GFAP. The grafted rMSCs expressed BDNF abundantly in Ang - rMSCs group. These grafted rMSCs survived up to 3 months at least. No malignant sign was observed in these grafted cells. CONCLUSION : Ang - 1 gene - modified MSCs transplantation has better therapeutic efficacy in cerebral infarction than that of MSC transplantation. The transplantation of cells with gene engineering is an effective therapeutic method for stroke patients.
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第2期241-247,共7页 Chinese Journal of Pathophysiology
基金 国家卫生部科学研究基金-福建省卫生教育联合攻关计划资助项目(No.WKJ2005-2-011) 福建省自然科学基金计划高校专项资助项目(No.C0540003)
关键词 骨髓间质干细胞 慢病毒载体 血管生成素-1 脑梗死 Bone marrow mesenchymal stem cells Lentiviral vector Angiopoietin- 1 Brain infarction
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参考文献16

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