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目的:探讨miR-146a对小鼠单核-巨噬细胞株RAW264.7以及小鼠原代腹腔巨噬细胞Thl/Th2类细胞因子表达的影响。方法:体外培养的小鼠单核-巨噬细胞系RAW264.7细胞和腹腔新鲜分离的巨噬细胞分别瞬时转染miR-146a mimics、阴性对照mimics(NC mimics)、抑制性miR-146a(miR-146a inhibitor)和抑制性miR-146a阴性对照(NC in-hibitor)。转染后,利用real time PCR定量检测IL-18、IL-5和IL-10的表达情况。结果:RAW264.7细胞和原代腹腔巨噬细胞在转染miR-146a mimics后IL-18的表达水平明显降低(P<0.05),转染miR-146a inhibitor后IL-18的表达水平明显升高(P<0.05),但对IL-5和IL-10的表达,两种转染形式均没有影响。结论:巨噬细胞RAW264.7及原代巨噬细胞表达的miR-146a能够负向调控Thl类细胞因子IL-18的表达,但是不能调控Th2类细胞因子IL-5及IL-10的表达。
Objective: To investigate the effect of miR-146a on the expression of mouse RAW264.7 monocyte-macrophage cell line and the expression of Th1 / Th2 cytokines in mouse peritoneal macrophages. METHODS: Mouse monocyte-macrophage cell line RAW264.7 cells and fresh peritoneal macrophages were transiently transfected with miR-146a mimics, negative control mimics (NC mimics) and miR-146a -146a inhibitor) and inhibitory miR-146a negative control (NC in-hibitor). After transfection, the expression of IL-18, IL-5 and IL-10 was quantified by real time PCR. Results: The expression of IL-18 in RAW264.7 cells and primary peritoneal macrophages was significantly decreased after transfected with miR-146a mimics (P <0.05), and the expression of IL-18 was significantly increased after transfected with miR-146a inhibitor High (P <0.05), but had no effect on the expression of IL-5 and IL-10 in both transfection modes. Conclusion: Macrophage RAW264.7 and miR-146a expressed by primary macrophages can negatively regulate the expression of Th1 cytokine IL-18, but can not regulate the expression of Th2 cytokines IL-5 and IL-10.