| 自杀基因ePNP诱导小鼠巨噬细胞凋亡的研究 |
| 投稿时间：2017-03-12 修订日期：2017-04-01 点此下载全文 |
| 引用本文：于敏敏,尹洪超.自杀基因ePNP诱导小鼠巨噬细胞凋亡的研究[J].医学研究杂志,2018,47(7):21-26 |
| DOI：
10.11969/j.issn.1673-548X.2018.07.006 |
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| 基金项目:国家高技术研究发展计划（"863"计划）项目（81670430） |
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| 中文摘要:目的 通过制作ePNP转基因小鼠，研究自杀基因ePNP对小鼠巨噬细胞凋亡的影响。方法 实验分ePNP转基因小鼠组和野生型小鼠组，在体外培养的各组小鼠腹腔巨噬细胞中分别加入浓度为0、0.125、0.25、0.5、1.0和2.0μg/ml的前体药物氟达拉滨（fludarabine），采用活细胞计数试剂盒（CCK-8）检测腹腔巨噬细胞加药后的细胞存活率；其次，给ePNP转基因小鼠和野生型小鼠腹腔注射剂量分别为0、10、20、60mg/kg的fludarabine，分别采用流式细胞术和原位凋亡检测试剂盒（TUNEL法）检测加入fludarabine后腹腔巨噬细胞和组织（肺、脾）巨噬细胞的凋亡情况。结果 CCK-8检测显示给药后ePNP小鼠腹腔巨噬细胞存活率明显下降，尤其在浓度为2.0μg/ml，细胞存活率仅为10.8%±0.8%；其次，流式细胞术和TUNEL结果也显示给药后其腹腔巨噬细胞和肺、脾巨噬细胞凋亡明显增加，并且存在剂量效应关系。CCK-8、流式细胞术和TUNEL法均显示野生型小鼠给药后没有明显巨噬细胞凋亡。结论 自杀基因ePNP能在前体药物fludarabine协同作用下，诱导小鼠巨噬细胞凋亡。 |
| 中文关键词:自杀基因 巨噬细胞凋亡 ePNP 氟达拉滨 |
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| Effect of E.coli Purine Nucleoside Phosphorylase on Inducing the Apoptosis of Murinemacrophages |
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| Abstract:Objective To investigate the effect of the suicide gene E. coli purine nucleoside phosphorylase (ePNP) on mouse macrophages apoptosis by using transgenic mice models. Methods The cell survival rate of peritoneal macrophages in ePNP transgenic mice and wide-type mice were detected by Cell Counting Kit-8(CCK-8)after adding prodrug (fludarabine) with the dosage of 0,0.125,0.25,0.5,1.0 and 2.0μg/ml respectively in vitro. The apoptosis of peritoneal macrophages and tissue(lung, spleen)macrophages were detected by flow cytometry and TUNEL assay after injecting fludarabine intraperitoneally with the dosage of 0, 10, 20 and 60mg/kg respectively. Results CCK-8 assay demonstrated that the survival rate of peritoneal macrophages of ePNP transgenic mice was obviously decreased after the treatment. Cell survival rate was only 10.8%±0.8% with the dosage of 2.0μg/ml. The flow cytometry showed that the peritoneal macrophages apoptosis of transgenic mice was significantly increased with a dose-effect relationship after administration, while the counterpart of wild-type mice did not change. Likewise, TUNEL assay showed that the lung and spleen macrophages apoptosis of transgenic mice were significantly increased with a dose-effect relationship after administration, while the counterpart of wild-type mice did not change. Conclusion The suicide gene ePNP can induce the apoptosis of mouse macrophages with synergistic action of fludarabine. |
| keywords:Suicide gene Macrophage apoptosis ePNP Fludarabine |
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