阳离子脂质体介导神经营养因子-3在大鼠雪旺细胞中的表达  

Expression of cationic liposome-mediated neurotropin-3 in Schwann cell

作　　者：施新革[1] 宗海斌[2] 董玉珍[1] 侯文根[1] 梁秋冬[1] 

机构地区：[1]新乡医学院第一附属医院骨科,河南卫辉453100 [2]新乡医学院基础医学院技能实验室

出　　处：《军医进修学院学报》2011年第9期956-958,964,共4页Academic Journal of Pla Postgraduate Medical School

基　　金：河南省教育厅自然科学研究计划项目(2010A320010)

摘　　要：目的观察阳离子脂质体转染神经营养因子-3(Neurotrophin-3,NT-3)基因在大鼠雪旺细胞(schwann cell,SC)的表达和提高SC细胞分泌NT-3的能力。方法采用阳离子脂质体介入法,将脂质体转染酶介导的NT-3转染至SC细胞,以转染后及未转染作为实验组和对照组,于转染后1、2、4、8周用ELISA双抗体夹心法测定NT-3蛋白的表达,采用DNA酶切鉴定转染后NT-3的DNA,免疫组化S-100染色法检测转染前后SC纯度。电镜下观察转染后SC细胞结构。结果转染后2、4、8周NT-3蛋白表达量与转染前比较差异有统计学意义(P<0.05)。转染后NT-3的DNA酶切鉴定结果与NT-3基因片段相符。转染前后SC纯度分别为(94.1±2.3)%及(95.8±2.1)%,差异有统计学意义(P<0.05)。结论 NT-3基因可转染培养的SC并高效表达。SC作为受体细胞易于获取并能在体外大量培养繁殖,能较长时间稳定大量表达所携带基因而不衰减。Objective To observe the expression of cationic liposome-transfected neurotrophin-3（NT-3） gene in Schwann cells（SC） and increase the ability of SC to secret NT-3.Methods Liposome-mediated NT-3 was transfected into SC by cationic liposome intervention and divided into experimental group and control group.NT-3 expression was detected in SC with the ELISA double antibody sandwich method 1,2,4,and 8 weeks after transfection.DNA of SC was identified by enzyme digestion.Purity of SC was detected with immunohistology S-100 staining.Structure of transfected NT-3 was observed under an electron microscope.Results The expression level of NT-3 in SC was significantly higher 2,4 and 8 weeks after transfection than before transfection（P〈0.05）.Enzyme digestion showed that the DNA level in transfected NT-3 gene was consistent with that in NT-3 gene fragments.The purity of SC was（94.1±2.3）% and（95.8±2.1）%,respectively,before and after transfection（P〈0.05）.Conclusion The NT-3 gene can transfect cultured SC and highly expressed in them.As receptor cells,SC can be easily obtained and massively cultured in vitro.

关 键 词：神经营养因子3 雪旺细胞 基因 

分 类 号：R34[医药卫生—基础医学]