布鲁氏菌对RAW264.7细胞内质网应激与细胞因子分泌的影响

发布时间：2018-10-26 14:58

【摘要】：【目的】布鲁氏菌与宿主相互作用的分子机制是目前的研究热点之一,布鲁氏菌通过形成来自于内质网的布氏小体而在巨噬细胞内生存和增殖,其机制目前尚不清楚,宿主细胞内质网应激反应对病原感染和炎症的调控密切相关。揭示内质网应激反应在布鲁氏菌感染巨噬细胞中的作用以及布鲁氏菌感染对巨噬细胞分泌免疫因子的影响。【方法】构建布鲁氏菌感染RAW264.7模型,在感染后不同时间收集细胞,通过实时荧光定量PCR检测细胞内质网应激反应标志分子GRP78和CHOP,以及TNF-α、IL-1β和IL-6在m RNA水平的变化;通过Western blot和ELISA分别检测其蛋白水平的变化。【结果】布鲁氏菌感染RAW264.7细胞的最佳感染复数MOI为100?1;证明在布鲁氏菌感染4-6 h,巨噬细胞可杀伤侵入的布鲁氏菌,之后存活的细菌可在细胞内增殖;感染后24 h出现细胞凋亡,48 h出现大量细胞坏死。布鲁氏菌感染可激活RAW264.7细胞的内质网应激反应,促进GRP78的表达,同时,抑制免疫因子的分泌。【结论】内质网应激反应参与了RAW264.7对布鲁氏菌感染的调节。
[Abstract]:[objective] the molecular mechanism of the interaction between brucella and its host is one of the hot topics. Brucella survives and proliferates in macrophages through the formation of Brucella bodies from the endoplasmic reticulum. The endoplasmic reticulum stress response of host cells is closely related to the regulation of pathogenic infection and inflammation. To reveal the role of endoplasmic reticulum stress in brucella infection with macrophages and the effect of brucella infection on the secretion of immune factors by macrophages. [methods] the RAW264.7 model of brucella infection was established. The changes of GRP78 and CHOP, TNF- 伪, IL-1 尾 and IL-6 at the level of m RNA were detected by real-time fluorescence quantitative PCR. The protein level was detected by Western blot and ELISA. [results] the best complex number MOI of Brucella infection in RAW264.7 cells was 100g / 1; The results showed that the macrophages could kill the invasive brucellosis for 4-6 hours, and the surviving bacteria could proliferate in the cells, apoptosis appeared at 24 h after infection, and a large number of cell necrosis occurred at 48 h after infection. Brucella infection can activate the endoplasmic reticulum stress response of RAW264.7 cells, promote the expression of GRP78, and inhibit the secretion of immune factors. [conclusion] endoplasmic reticulum stress response is involved in the regulation of brucella infection by RAW264.7.
【作者单位】：西北农林科技大学动物医学院农业部动物生物技术重点实验室;
【基金】：国家自然科学基金项目(No.31672584) 陕西省科技统筹创新工程计划项目(No.2016TZC-N-13-5)~~
【分类号】：S852.614

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