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碱性成纤维细胞生长因子基因转染大鼠骨髓间充质干细胞的基因表达 被引量：4

Expression of the basic fibroblast growth factor gene after being transfected into rat bone marrow mesenchymal stem cells
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摘要 目的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)是一类存在于骨髓等多种组织中的具有多向分化潜能的组织干细胞。为研究碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)转染BMSCs对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)的修复作用及机制,将b FGF基因转染至体外培养的大鼠BMSCs,测定b FGF基因表达情况。方法分离、培养并鉴定BMSCs,构建重组质粒b FGF-pc DNA3.1并测序。脂质体介导b FGF-pc DNA3.1转染BMSCs为b FGF-pc DNA3.1质粒转染组,脂质体介导pc DNA3.1转染BMSCs为空白pc DNA3.1质粒转染组,未转染的BMSCs为未转染组,G418筛选14d,qRT-PCR、Western blot检测b FGF基因及蛋白的表达。结果序列测定证实与Gen Bank报道的b FGF全长基因序列一致。b FGF-pc DNA3.1质粒转染组b FGF基因表达(7.028±0.568)较空白pc DNA3.1质粒转染组(1.000±0.082)及未转染组(1)明显升高(P<0.01),而pc DNA3.1质粒转染组和未转染组相比差异无统计学意义(P>0.05)。b FGF-pc DNA3.1质粒转染组b FGF蛋白表达(1.017±0.054)较空白pc DNA3.1质粒转染组(0.217±0.009)及未转染组(0.165±0.013)明显升高(P<0.05),而空白pc DNA3.1质粒转染组及未转染组b FGF蛋白表达差异无统计学意义(P>0.05)。结论脂质体介导下,重组真核表达载体b FGF-pc DNA3.1能被转入BMSCs,且表达了b FGF基因及蛋白。 Objective Bone marrow mesenchymal stem cells（ BMSCs）,a kind of stem cells with multiple differentiation potentials,exist in the bone marrow and other organizations. This study aimed to investigate the repairing effect of the exogenous basic fibroblast growth factor（ b FGF） against chronic obstructive pulmonary disease（ COPD） and its action mechanism,and to determine the expression of the b FGF gene in transfected rat BMSCs. Methods BMSCs were isolated,cultured and identified. The recombinant plasmid b FGF-pc DNA3. 1 was constructed and sequenced. Liposome-mediated b FGF-pc DNA3. 1 plasmid was transfected into the BMSCs of the rat（ b FGF-pc DNA3. 1 transfection group）,liposome-mediated pc DNA3. 1 transfected into the BMSCs（ pc DNA3. 1 transfection group）,and untransfected BMSCs used as the control. G418 screening was performed for 14 days. The gene and protein expressions of b FGF were determined by qRT-PCR and Western blot. Results The full-length sequence of the b FGF gene was consistent with that of the Gen Bank. The expression of the b FGF gene was significantly higher in the b FGF-pc DNA3. 1 transfection group（ 7. 028 ± 0. 568） than in the pc DNA3. 1 transfection group（ 1. 000 ± 0. 082） and the non-transfection control（ 1）（ P〈0. 01）,but with no statistically significant difference between the latter two groups（ P〉0. 05）. The expression of the b FGF protein was also remarkably higher in the b FGF-pc DNA3. 1 transfection group（ 1. 017 ± 0. 054） than in the pc DNA3. 1 transfection group（ 0. 217 ±0. 009） and the non-transfection control（ 0. 165 ± 0. 013）（ P〈0. 05）,with no statistically significant difference between the latter two groups（ P〉0. 05）. Conclusion Mediated by the liposome reagent,the recombinant eukaryotic expression vector b FGF-pc DNA3. 1 can be transfected into rat BMSCs and expresses the b FGF gene and protein.
作者 赵悦 马琳 彭珊珊 淦鑫
机构地区 南昌大学第一附属医院呼吸科
出处 《医学研究生学报》 CAS 北大核心 2015年第12期1246-1251,共6页 Journal of Medical Postgraduates
基金 国家自然科学基金(81260004)
关键词 碱性成纤维细胞生长因子 骨髓间充质干细胞 慢性阻塞性肺疾病 基因表达 Basic fibroblast growth factor Bone marrow mesenchymal stem cells Chronic obstructive pulmonary diseases Gene expression
分类号 R563.9 [医药卫生—呼吸系统]
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参考文献21

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共引文献4

同被引文献51

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医学研究生学报
2015年 第12期

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