布鲁氏菌OMP16对RAW264.7细胞凋亡与免疫活性的影响

doi:10.11843/j.issn.0366-6964.2022.08.022

摘要/Abstract

摘要： 旨在研究布鲁氏菌外膜蛋白OMP16对小鼠巨噬细胞RAW264.7细胞凋亡和免疫活性的影响及其机制探讨。以OMP16处理RAW264.7细胞为模型，通过MTT以及Hoechst法检测OMP16对细胞活力的影响，通过Western blot检测Caspase-3、GRP78、CHOP的表达情况，流式细胞术检测OMP16对细胞凋亡的影响以及RT-PCR检测免疫因子IL-1β、IL-6和TNF-α的变化。结果表明，OMP16能够影响RAW264.7细胞的活力，且有浓度依赖性；添加OMP16后能够显著增加凋亡标志因子Caspase-3的表达（P<0.001），并促进RAW264.7细胞的凋亡；作用24、36 h之后，显著引起内质网应激标志蛋白GRP78、CHOP的表达；并且能显著上调IL-1β、IL-6、TNF-α等炎性因子的mRNA表达水平。布鲁氏菌OMP16能够诱导RAW264.7凋亡，OMP16显著引起内质网应激CHOP通路的激活。

关键词: 布鲁氏菌, 外膜蛋白, OMP16, RAW264.7, 细胞凋亡, 免疫活性

Abstract: The purpose of this study was to study the effect of Brucella outer membrane protein 16 on apoptosis and immune activity of mouse macrophage RAW264.7 cell, and to explore its mechanism. In this study, OMP16 treated RAW264.7 cells were used as a model to detect the effects of OMP16 on cell viability by MTT and Hoechst assay. The expressions of Caspase-3, GRP78 and CHOP level were detected by Western blot. The apoptosis of OMP16 treated RAW264.7 was detected by flow cytometry. The changes of IL-1β, IL-6 and TNF-α were detected by RT-PCR. The results showed that OMP16 could affect the activity of RAW264.7 cells in a concentration-dependent manner. OMP16 significantly increased the expression of caspase-3 (P<0.001) and promoted apoptosis of OMP16 treated RAW264.7 cells. The expression of ER stress marker protein GRP78 and CHOP was significantly increased in RAW264.7 cells after OMP16 treated 24 and 36 h. The mRNA expression levels of IL-1β, IL-6, TNF-α were significantly up-regulated. Brucella OMP16 can induce RAW264.7 apoptosis, and OMP16 significantly causes ER stress through CHOP pathway.

Key words: Brucella, outer membrane protein, OMP16, RAW264.7, apoptosis, immune activity

中图分类号:

S852.61

李杨, 周栋, 尹彦龙, 张广冻, 相彩霞, 支飞杰, 白芙蓉, 林鹏飞, 靳亚平, 王爱华. 布鲁氏菌OMP16对RAW264.7细胞凋亡与免疫活性的影响[J]. 畜牧兽医学报, 2022, 53(8): 2642-2651.

LI Yang, ZHOU Dong, YIN Yanlong, ZHANG Guangdong, XIANG Caixia, ZHI Feijie, BAI Furong, LIN Pengfei, JIN Yaping, WANG Aihua. Effects of Brucella Outer Membrane Protein 16 on Apoptosis and Immune Activity of RAW264.7 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2642-2651.

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