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目的：建立一个在大鼠心肌细胞表达人白细胞介素１０（ＨｕｍａｎＩｎｔｅｒｌｅｕｋｉｎ１０，ｈＩＬ１０）的重组逆转录病毒载体基因转移系统，为下一步的研究工作做准备。方法：将构建的表达人白细胞介素１０的逆转录病毒重组体ｐＬＸ（ｈＩＬ１０）ＳＮ经ＰＡ３１７细胞包装，Ｇ４１８筛选，ＮＩＨ３Ｔ３细胞进行病毒滴度测定，选滴度最高的克隆（６×１０５ＣＦＵ／ｍｌ）作为感染大鼠心肌细胞的感染细胞；用重组逆转录病毒感染大鼠心肌细胞，应用聚合酶链反应（ＰＣＲ）及反转录聚合酶链反应（ＲＴＰＣＲ）检测转染大鼠心肌细胞ＤＮＡ及ｍＲＮＡ，应用ＥＬＩＳＡ测定ｈＩＬ１０在心肌细胞的表达。结果：外源性ｈＩＬ１０基因已整合到心肌细胞染色体ＤＮＡ并有效地转录和翻译，表达水平最高达１７５５ｎｇ／（１０６ｃｅｌｓ·２４ｈ）。结论：外源性ｈＩＬ１０基因可以转移到大鼠心肌细胞并稳定表达，为今后研究ｈＩＬ１０的作用机制及在治疗疾病中的应用打下了基础。
OBJECTIVE: To establish a recombinant retroviral vector gene transfer system expressing human Interleukin 10 (hIL10) in rat cardiomyocytes, and to prepare for the further research work. Methods: The retrovirus recombinant pLX (hIL10) SN expressing human interleukin 10 was packaged by PA317 cells and screened by G418. NIH3T3 cells were used for virus titer determination. The highest titer of clone (6 × 105CFU / Ml) were used as infected cells in rat cardiomyocytes infected with cardiomyocytes. Rat cardiomyocytes were infected with recombinant retrovirus and transfected into rats by polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) Cardiomyocytes DNA and mRNA, the application of ELISA assay hIL 10 expression in cardiomyocytes. Results: The exogenous hIL10 gene was integrated into the chromosomal DNA of cardiomyocytes and transcribed and translated efficiently. The expression level reached 1755ng / (106cels · 24h). Conclusion: Exogenous hIL10 gene can be transferred to rat cardiac myocytes and stably expressed, laying a foundation for future study of the mechanism of hIL10 and its application in the treatment of diseases.