Effect of overexpression of endothelial nitric oxide synthase traffic inducer on human umbilical vein endothelial cells
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF PERINATAL MEDICINE
- Issue:
- 1
- DOI:
- 10.3760/cma.j.issn.1007-9408.2007.01.005
- Key Word:
- 胞间信号肽类和蛋白质类;一氧化氮合酶;脐静脉;内皮,血管;转染
Abstract： 目的 研究人内皮型一氧化氮合酶运输介导物(endothelial nitric oxide synthase traffic inducer, NOSTRIN)基因在体外培养的人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVEC)中的过度表达对该细胞生物学特性的影响.方法 脂质体介导转染体外培养的HUVEC,G418筛选阳性克隆.免疫组织化学法检测第八因子相关抗原(FⅧRag)的表达;细胞计数法绘制细胞生长曲线以观察细胞的增殖能力;Western 印迹检测细胞中NOSTRIN和eNOS蛋白质的表达;分光光度法和硝酸还原酶法分别测定细胞培养上清中eNOS的活性和NO代谢产物亚硝酸基/亚硝基(NO2-/NO3-)水平;光镜、电镜观察细胞的形态和超微结构.结果 转染前后细胞均为血管内皮来源细胞;转染NOSTRIN基因后细胞的增殖能力明显降低;Western印迹显示细胞转染NOSTRIN基因后其表达量明显增高,而eNOS蛋白质的表达量在各组细胞之间无差异(P＞0.05);转染NOSTRIN的细胞培养上清中eNOS活性[(11.727±3.09) U/ml]与转染空载体细胞[(22.69±3.36) U/ml]和对照组细胞[(21.85±3.47) U/ml]比较降低显著(P＜0.01),同时转染NOSTRIN细胞培养上清中 NO2-/NO3-[(25.56±2.49) μmol/L],与转染空载体细胞[(48.37±2.61) μmol/L]和对照组[(49.06±2.78) μmol/L]比较显著降低(P＜0.01);光镜和电镜观察看到转染NOSTRIN后对HUVEC有损伤作用(HUVEC拉长、微绒毛变短、细胞膜损伤,胞浆内有脂滴空泡,核膜不完整).结论 NOSTRIN的高表达影响了HUVEC的生物学特性,对HUVEC有明显的损伤作用.
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