MRSA对巨噬细胞系RAW264.7自噬的影响

发布时间：2018-02-27 05:15

本文关键词： 耐甲氧西林金黄色葡萄球菌巨噬细胞自噬　出处：《新医学》2016年05期 　论文类型：期刊论文

【摘要】：目的探讨耐甲氧西林金黄色葡萄球菌(MRSA)感染对于巨噬细胞自噬的影响。方法以小鼠单核巨噬细胞系RAW264.7为研究对象,以革兰阳性菌MRSA感染3 h组作为实验组,设立空白对照组(对照组),另设MRSA感染0、30、60、120、150、180 min组。以MRSA感染细胞后,荧光定量PCR检测自噬相关基因5(ATG5)、ATG7、ATG12的表达情况,蛋白免疫印迹法检测自噬相关蛋白Beclin1及微管相关蛋白轻链3(LC3)的表达情况。结果感染MRSA 3 h后,实验组RAW264.7细胞中ATG5、ATG7、ATG12的mRNA表达水平均高于对照组(P均0.01),LC3蛋白相对表达量也比对照组增加(P0.01),2组间Beclin1蛋白相对表达量比较差异无统计学意义(P0.05)。ATG7、ATG12 mRNA和LC3及Beclin1蛋白相对表达量均在感染MRSA 180 min组最高,ATG5 mRNA在感染MRSA 150 min组最高。结论 MRSA可以诱导巨噬细胞发生自噬,自噬基因激活存在先后顺序,自噬可能为巨噬细胞吞噬胞外菌MRSA的另一种免疫灭活形式。
[Abstract]:Objective to investigate the effect of methicillin-resistant staphylococcus aureus (MRSA) infection on macrophage autophagy. Methods Murine mononuclear macrophage cell line RAW264.7 was used as the study object and Gram-positive bacteria MRSA as experimental group for 3 h. A blank control group (control group) was established. After MRSA infected cells were infected with MRSA, the expression of ATG7 / ATG12 was detected by fluorescence quantitative PCR. The expression of autophagy associated protein (Beclin1) and microtubule-associated protein light chain (3LC-3) were detected by Western blot. The mRNA expression of ATG5, ATG7, ATG12 in RAW264.7 cells in experimental group was higher than that in control group (P = 0.01). There was also no significant difference in the relative expression of Beclin1 protein between the two groups. There was no significant difference in the relative expression of Beclin1 protein, LC3 and Beclin1 between the two groups. The highest amount of ATG5 mRNA was found in the infected MRSA 180 min group and the highest in the MRSA 150 min group. Conclusion MRSA can induce autophagy in macrophages. Autophagy gene activation exists in sequence, autophagy may be another form of immune inactivation of macrophage phagocytic extracellular bacteria MRSA.
【作者单位】：中山大学附属第三医院呼吸内科内科ICU;
【基金】：国家自然科学基金面上项目(81170004)
【分类号】：R515

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