【摘要】：目的探討miR-301a對(duì)巨噬細(xì)胞炎癥因子表達(dá)水平的影響,從microRNA角度闡明動(dòng)脈粥樣硬化的發(fā)病機(jī)制,為動(dòng)脈粥樣硬化的防治提供新的思路。方法高脂喂養(yǎng)Apo E-/-小鼠建立動(dòng)脈粥樣硬化模型,收集小鼠主動(dòng)脈血管,利用real-time PCR檢測(cè)組織中miR-301a的表達(dá)水平;利用脂質(zhì)體在RAW264.7細(xì)胞中轉(zhuǎn)染miR-301a mimic和miR-301a inhibitor,用real-time PCR檢測(cè)細(xì)胞中miR-301a水平,并檢測(cè)腫瘤壞死因子α(TNF-α)、白細(xì)胞介素6(IL-6)和單核細(xì)胞趨化蛋白1(MCP-1)的表達(dá)水平,用Western blot檢測(cè)NF-κB抑制因子(NKRF)蛋白水平,用免疫熒光染色檢測(cè)p65細(xì)胞定位。結(jié)果在高脂喂養(yǎng)的Apo E-/-小鼠主動(dòng)脈血管壁組織中miR-301a的表達(dá)水平升高;在RAW264.7細(xì)胞中過(guò)表達(dá)miR-301a可抑制NKRF蛋白水平,促進(jìn)p65細(xì)胞核定位,升高細(xì)胞中TNF-α、IL-6和MCP-1的mRNA表達(dá);在RAW264.7細(xì)胞中低表達(dá)miR-301a可升高NKRF蛋白水平,促進(jìn)p65細(xì)胞質(zhì)定位,減少細(xì)胞中TNF-α、IL-6和MCP-1的mRNA表達(dá)。結(jié)論在高脂喂養(yǎng)的Apo E-/-小鼠主動(dòng)脈血管壁組織中miR-301a表達(dá)水平升高;在RAW264.7細(xì)胞中miR-301a通過(guò)調(diào)節(jié)NKRF的表達(dá)影響p65活性進(jìn)而調(diào)控TNF-α、IL-6和MCP-1的mRNA表達(dá),提示microRNA在動(dòng)脈粥樣硬化發(fā)病的炎癥機(jī)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- 偽 (TNF- 偽), 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- 偽 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- 偽 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- 偽 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.
【作者單位】： 北京醫(yī)院國(guó)家老年醫(yī)學(xué)中心衛(wèi)生部老年醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(81600618) 北京醫(yī)院博士啟動(dòng)基金項(xiàng)目(BJ-2015-106)
【分類(lèi)號(hào)】：R543.5

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