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    李木楠, 李双双, 孟凡爽, 等. 抑制miR-29a-3p表达对小鼠巨噬细胞凋亡的影响及机制[J]. 华南农业大学学报, 2018, 39(1): 64-69. DOI: 10.7671/j.issn.1001-411X.2018.01.011
    引用本文: 李木楠, 李双双, 孟凡爽, 等. 抑制miR-29a-3p表达对小鼠巨噬细胞凋亡的影响及机制[J]. 华南农业大学学报, 2018, 39(1): 64-69. DOI: 10.7671/j.issn.1001-411X.2018.01.011
    shu
    LI Munan, LI Shuangshuang, MENG fanshuang, HAN Peng, ZHANG Wenhui, ZHANG Linbo. Mechanism and influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage[J]. Journal of South China Agricultural University, 2018, 39(1): 64-69. DOI: 10.7671/j.issn.1001-411X.2018.01.011
    Citation: LI Munan, LI Shuangshuang, MENG fanshuang, HAN Peng, ZHANG Wenhui, ZHANG Linbo. Mechanism and influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage[J]. Journal of South China Agricultural University, 2018, 39(1): 64-69. DOI: 10.7671/j.issn.1001-411X.2018.01.011
    shu

    抑制miR-29a-3p表达对小鼠巨噬细胞凋亡的影 响 及 机 制

    Mechanism and influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage

      摘要
    • 摘要:
      目的  研究miR-29a-3p下调表达对小鼠巨噬细胞凋亡及相关基因表达的影响，为探讨结核病发病机制、研发新的诊断与治疗方法提供依据。
      方法  将重组抑制载体pEZX-AM02-miR-29a-3p转染RAW264.7细胞，24、36和48 h荧光显微镜观察转染效率，Real time-PCR法检测miR-29a-3p及凋亡相关基因caspase3、caspase7、caspase8、Bcl-2、Mcl-1和Bax的表达水平，流式细胞术检测RAW264.7细胞的凋亡率。
      结果  重组抑制载体转染后，细胞内miR-29a-3p表达水平明显下降；而caspase7、caspase8、Bcl-2、Mcl-1和Bax等基因的表达出现不同程度的上调；流式细胞术分析发现随着转染时间延长，细胞凋亡率增加，且36 h凋亡率最高。
      结论  pEZX -AM02-miR-29a-3p重组载体可抑制小鼠巨噬细胞中miR-29a-3p的表达，通过靶向上调caspase7、caspase8、Bcl-2和Mcl-1等基因的表达促进巨噬细胞的凋亡。

       

      Abstract:
      Objective  To study the influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage and expression of apoptosis-associated genes, and provide references for investigating the pathogenesis mechanism of mycobacterium tuberculosis (MTB) and developing new diagnosis and treatment methods.
      Method  The recombinant inhibitor vector pEZX-AM02-miR-29a-3p was transfected into RAW264.7 cells. Transfection efficiency was observed through fluorescence microscopy, the expression levels of miR-29a-3p and apoptosis-associated genes caspase3, caspase7, caspase8, Bcl-2, Mcl-1 and Bax were detected by RT-PCR, and the apoptosis rates of RAW264.7 cells were detected by flow cytometry at 24, 36, 48 h after transfection.
      Result  After transfected with pEZX-AM02-miR-29a-3p recombinant inhibitor vector, miR-29a-3p expression in RAW264.7 cells decreased significantly, while the target genes caspase7, caspase8, Bcl-2, Mcl-1 and Bax were up-regulated at different levels. The results of flow cytometry indicated that the apoptosis rates increased as the transfection time progressed and reached the peak at 36 h.
      Conclusion  The expression of miR-29a-3p in mouse macrophage are inhibited by transfecting with pEZX-AM02-miR-29a-3p, which promotes the apoptosis of macrophage by up-regulating the expression of target genes, suah as caspase7, caspase8, Bcl-2 and Mcl-1.

       

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