摘要
背景:血管舒缓素即组织激肽释放酶1,是组织激肽释放酶-激肽系统重要组成部分之一。有研究表明血管舒缓素通过促进血管新生,抑制心肌炎症产生等发挥其对心血管系统的保护作用,但并未从干细胞诱导分化方面进行探讨。目的:实验利用已构建的腺病毒载体,将其携带的血管舒缓素基因转染到大鼠骨髓间充质干细胞上,观察其是否成功转染及转染率如何。方法:以腺病毒作为载体,将目的基因血管舒缓素转染到大鼠骨髓间充质干细胞上,并采用荧光显微镜、四甲基偶氮唑盐法和流式细胞技术观察病毒对细胞的转染效果,以确定最佳的病毒感染复数。结果与结论:荧光显微镜下观察到腺病毒携带的血管舒缓素目的基因成功转染到大鼠骨髓间充质干细胞上;流式细胞仪结果显示转染率跟病毒感染复数值的大小有关系,当病毒感染复数为150时,转染率为80.8%;四甲基偶氮唑盐法结果提示,病毒感染复数为200时,细胞生长受到明显的抑制。结果证实,腺病毒介导的血管舒缓素能成功转染到大鼠骨髓间充质干细胞上,且最佳的病毒感染复数为150。
BACKGROUND： Kallikrein 1 is an important component of the kallikrein-kinin system. Studies have shown that kallikrein can protect the cardiovascular system by promoting angiogenesis and inhibiting myocardial inflammation, but there is no report on its effect on inducing differentiation of stem cells.OBJECTIVE： To determine the transfection efficiency of kallikrein 1 adenoviral vector in rat bone mesenchymal stem cells. METHODS： Using adenovirus as a vector, the target gene kallikrein 1 was transfected into rat bone marrow mesenchymal stem cells. Fluorescence microscopy, MTT method and flow cytometry were used to investigate the effect of transfection and determine the optimal multiplicity of infection. RESULTS AND CONCLUSION： Adenovirus carrying kallikrein 1 was successfully transfected into rat bone marrow mesenchymal stem cells. Results from flow cytometry showed that the transfection efficiency was associated with the multiplicity of infection. When the multiplicity of infection was 150, the transfection efficiency was 80.8%. MTT results showed that when the multiplicity of infection was 200, the cell growth was inhibited remarkably. These findings indicate that adenovirus-mediated kallikrein 1 can be successfully transfected into rat bone marrow mesenchymal stem cells with the optimal multiplicity of infection=150.
出处
《中国组织工程研究》
CAS
北大核心
2015年第45期7249-7253,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81260597)
甘肃省高校基本科研业务费项目(2012-3)~~
作者简介
贾佳，女，1988年生，汉族，甘肃中医药大学在读硕士，主要从事中医药治疗心血管疾病的研究。
通讯作者：胡继宏，博士，副教授，硕士生导师，甘肃中医药大学基础医学院，甘肃省兰州市730000
