摘要
目的探讨转染内皮型一氧化氮合酶(eNOS)基因后内皮细胞(EC)的功能变化。方法采用脂质体法转染eNOS基因于实验犬EC;RT-PCR和免疫组化法检测转染效果;分别采用比色法和酶联免疫法检测细胞培养液一氧化氮合酶(NOS)、一氧化氮(NO)和vWF的浓度;并观察转染细胞生长增殖情况。结果RT-PCR产物电泳和测序及免疫组化法检测证实转染效果满意;转染后EC培养液NOS和NO浓度在不同时间明显升高(120h分别为33.53和32.99),与正常组对比差异显著(P<0.05)。转染后细胞生长增殖和vWF含量无显著差异。结论通过脂质体法成功地将eNOS基因转染于实验犬EC;转染后eNOS基因在mRNA和蛋白质水平均高效表达;内皮细胞eNOS活性显著增强;转染后细胞生物学功能稳定。
Objective To study endothelial nitric oxide synthase （eNOS） gene transfecting endothelial cells （ECs）. Methods eNOS gene was transfected into the steady ECs system by cationic liposomal transduction and check the transfection effect by RT-PCR and immunohistochemistry assay. To test the concentrations of NOS, nitric oxide （NO） and yon willebrand factor （vWF） in culture media by colorimetry and ELISA, respectively,and transfected EC function was observed. Results The effect of transfection was satisfactory with RT-PCR products electrophoresis, sequence and immunohistochemistry. The concentrations of NOS and NO in transfected EC culture media increased with time （P 〈 0.05） and reached 33.53 and 32.99 at the point of 120 h, respectively. There was no difference in the proliferation of transfected EC and the concentration of vWF between the groups of EC and eNOS. Conclusion eNOS gene can be successfully transfected into established steady ECs by cationic liposomal transduction. Transfected ECs have high overexpression at the mRNA and protein level, eNOS activity in transfected ECs is significantly higher. The biological functions of transfected ECs do not basically change.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2006年第3期278-281,共4页
Journal of Shanghai Jiao tong University:Medical Science
