miR-301a调节小鼠巨噬细胞中炎症因子的表达

发布时间：2018-07-17 14:44

【摘要】：目的探讨miR-301a对巨噬细胞炎症因子表达水平的影响,从microRNA角度阐明动脉粥样硬化的发病机制,为动脉粥样硬化的防治提供新的思路。方法高脂喂养Apo E-/-小鼠建立动脉粥样硬化模型,收集小鼠主动脉血管,利用real-time PCR检测组织中miR-301a的表达水平;利用脂质体在RAW264.7细胞中转染miR-301a mimic和miR-301a inhibitor,用real-time PCR检测细胞中miR-301a水平,并检测肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和单核细胞趋化蛋白1(MCP-1)的表达水平,用Western blot检测NF-κB抑制因子(NKRF)蛋白水平,用免疫荧光染色检测p65细胞定位。结果在高脂喂养的Apo E-/-小鼠主动脉血管壁组织中miR-301a的表达水平升高;在RAW264.7细胞中过表达miR-301a可抑制NKRF蛋白水平,促进p65细胞核定位,升高细胞中TNF-α、IL-6和MCP-1的mRNA表达;在RAW264.7细胞中低表达miR-301a可升高NKRF蛋白水平,促进p65细胞质定位,减少细胞中TNF-α、IL-6和MCP-1的mRNA表达。结论在高脂喂养的Apo E-/-小鼠主动脉血管壁组织中miR-301a表达水平升高;在RAW264.7细胞中miR-301a通过调节NKRF的表达影响p65活性进而调控TNF-α、IL-6和MCP-1的mRNA表达,提示microRNA在动脉粥样硬化发病的炎症机制中具有重要作用。
[Abstract]:Objective to investigate the effect of miR-301a on the expression of inflammatory cytokines in macrophages, to elucidate the pathogenesis of atherosclerosis from the microRNA perspective, and to provide a new idea for the prevention and treatment of atherosclerosis. Methods Atherosclerosis model was established by high-fat feeding Apo E-r-mice, the aortic vessels were collected, the expression of miR-301a was detected by real-time PCR, the miR-301a mimic and miR-301a inhibitor were transfected into RAW264.7 cells by liposome, and the miR-301a levels were detected by real-time PCR. The expression of tumor necrosis factor- 伪 (TNF- 伪), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), NF- 魏 B inhibitory factor (NKRF) protein and p65 cell localization were detected by Western blot and immunofluorescence staining. Results the expression of miR-301a was increased in the vascular wall of Apo E-r-mice fed with high fat, and the overexpression of miR-301a in RAW264.7 cells inhibited NKRF protein level, promoted the nuclear localization of p65, and increased the mRNA expression of TNF- 伪 IL-6 and MCP-1 in RAW264.7 cells. The low expression of miR-301a in RAW264.7 cells increased NKRF protein level, promoted the cytoplasmic localization of p65 cells, and reduced the mRNA expression of TNF- 伪 IL-6 and MCP-1 in RAW264.7 cells. Conclusion the expression of miR-301a is increased in the vascular wall of Apo E-r-mice fed with high fat, and the expression of TNF- 伪 IL-6 and MCP-1 in RAW264.7 cells is affected by the effect of miR-301a on the expression of p65 by regulating the expression of NKRF. These results suggest that microRNA plays an important role in the inflammatory mechanism of atherosclerosis.
【作者单位】： 北京医院国家老年医学中心卫生部老年医学重点实验室;
【基金】：国家自然科学基金项目(81600618) 北京医院博士启动基金项目(BJ-2015-106)
【分类号】：R543.5

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