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- Title
牛坏死杆菌对小鼠巨噬细胞的增殖和自噬影响的初步探究.
- Authors
王天硕; 于思雯; 毕栏; 赵鹏宇; 蒋凯; 肖佳薇; 郭东华
- Abstract
To investigate the effects of Fusobacterium necrophorum on the proliferation and autophagy of mouse macrophage cell lines (RAW264.7 cells), the RAW264.7 cells were infected with Fusobacterium necrophorum (strain A25) at a multiplicity of infection (MOI) of 10 for 1 hour, 2 hours and 4 hours as experimental group, RA W264. 7 cells without any treatment were used as the control group. The cell fluorescence was observed by EdU and MDC staining, and the cell proliferation rate and the number of autophagosomes in each group were calculated. EdU staining showed that the amount of green fluorescence decreased gradually with the prolongation of infection time, but control cells' green fluorescence did not change. Compared with the control group, the cell proliferation rate of the infection group significantly reduced with the prolongation of infection time (P<0.05). The results of MDC staining showed that compared with the control group, the green fluorescence gradually increased with the prolongation of infection time in the infection group, and the number of autophagosomes increased significantly with the prolongation of infection time compared with the control group. The above results showed that the infection of RA W264. 7 cells by Fusobacterium necrophorum inhibited its proliferation and induced autophagy. The transcription levels of autophagy genes Beclin-1 and P62 and the relative expression levels of autophagy proteins LC3 and P62 in the above groups were detected by fluorescence quantitative PCR (RT-qPCR) and western blot. RT-qPCR results showed no significant change in the relative mRNA transcription level of the Beclin-1 gene at l hour after infection compared with the control group. The relative mRNA transcription level of this gene increased significantly at 2 hours and 4 hours after infection (P<0.01 ), and the relative mRNA transcription level of P62 gene was significantly down-regulated at different infection times (P<0.001). Western blot results showed that compared with the control group, the expression ofLC3-II protein was significantly increased at different infection times (P<0.01), and the expression of P62 protein was significantly decreased at different infection times (P<0.001). The above results further demonstrated that Fusobacterium necrophorum induced autophagy in RA W264. 7 cells at the mRNA and protein expression levels of autophagy-related genes. This is also the first time that Fusobacterium necrophorum can induce autophagy in RA W264. 7 cells. This study provides an experimental basis for exploring the infection mechanism of Fusobacterium necrophorum.
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2023, Vol 45, Issue 9, p936
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202301010