摘要
为探究自噬作用对布鲁氏菌(Brucella)在巨噬细胞(Mφ)内增殖活力的影响,本研究将体外培养的Mφ细胞分组,分别采用自噬激活剂雷帕霉素(Ra)或抑制剂3-MA处理细胞,并在不同处理的细胞组中接种表达绿色荧光的重组猪种布鲁氏菌(rB.suis-GFP),在接种后0.5 h^24 h,一方面通过细胞中LC3-Ⅱ含量及Mφ内B.suis数量的测定,另一方面通过激光共聚焦显微镜观察LC3-Ⅱ与B.suis共定位情况。结果显示,感染B.suis后0.5 h,Mφ即可发生自噬作用,并且Ra+Mφ+B.suis组LC3-Ⅱ含量显著高于其它组,表明B.suis的感染能够促进Mφ内LC3-Ⅱ的形成,增强Mφ的自噬作用;另一方面,Ra+Mφ+B.suis组胞内B.suis数量显著高于3-MA+Mφ+B.suis组与Mφ+B.suis组,表明自噬作用能够促进B.suis在Mφ内的增殖;但随着胞内B.suis数量的增加,各组LC3-Ⅱ的水平却逐渐下降,表明B.suis感染Mφ数量达饱和后,Mφ虽然启动自噬作用杀灭了部分B.suis,但却不能完全抑制B.suis在Mφ内大量增殖。本研究为初步确定自噬在B.suis增殖过程中的作用提供了依据。
Abstract： In order to know the effect of autophagy on the replication of Brucella in Macrophage （Mφ）, Mφ was grouped firstly, then dealt with autophagy activator rapamycin and inhibitor 3-MA, and infected with GFP recombinant B.suis respectively in this study. 0.5 h to 24 h after inoculation, the LC3-II contents and B.suis quantity in cell culture were determinationed, and co-localization between LC3-II and B.suis were abserved by laser scanning confocal microscope. The result demonstrated that autophagy could be induced at 0.5 hours after B.suis infection in Mφ, and the LC3-II content in Ra＋Mφ＋B.suis group was significantly higher than that in other groups, indicating that B.suis infection could promote the formation of LC3-ll in Mφ and enhance the autophagy of Mφ. On the other hand, the quantity of intracellular B.suis in Ra＋Mφ＋B.suis group was significantly higher than that in 3-MA＋Mq~＋B.suis and Mq~＋B.suis groups, indicating that autophagy could promote the proliferation of B.suis in Mq~. However, with the number of intracellular B.suis increased, the levels of LC3-II in each group decreased gradually, which indicated when the number of B.suis to saturation, although Mφ could kill some B.suis by autophagy, it could not completely inhibit the proliferation of B.suis in Mφ. This study provided a basis for determining the role of autophagy in the process of B.suis
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2017年第12期1008-1012,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
山东省自然资金项目(ZR2013CQ005)
山东省现代农业产业技术体系创新团队项目(SDAIT-10-07)
关键词
自噬
布鲁氏菌
巨噬细胞
增殖
autophagy
Brucella
Macrophage （Mφ）
proliferation
