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卵形鲳鲹源创伤弧菌导致宿主细胞凋亡的作用机制
刘明珠1, 余 庆1, 肖贺贺2, 吴思婷3, 黎思巧1, 王一兵4, 聂振平1, 李鹏飞1
0
(1.广西科学院;2.河南师范大学;3.广西海洋生物技术重点实验室;4.广西民族大学海洋与生物技术学院)
摘要:
【目的】弧菌是自然界菌群中的一部分,作为一种有害菌不仅可以感染常见的水产品,有些弧菌更是对人体有一定的致病性。随着近些年水产养殖的不断扩大,由创伤弧菌(Vibrio vulnificus, V. vulnificus)引起的鱼病不胜枚举,不仅对养殖行业造成巨大的损失,同时对人类生命安全也存在着巨大威胁。本文对卵形鲳鲹源创伤弧菌感染宿主细胞的机制进行研究,目的是阐明创伤弧菌对宿主细胞的致死机理,为未来研发抗病菌功能产品提供理论依据。【方法】本文通过确定创伤弧菌胞外分泌物(extracellular bacteria products, ECPs)的蛋白浓度,以不同的蛋白浓度为基准,利用光镜观察和细胞活力CCK检测技术测定创伤弧菌ECPs对细胞的毒性。然后使用Hoechst染核观察及DNA Ladder来进一步确定其致死机制。【结果】根据细胞毒性结果,卵形鲳鲹源创伤弧菌ECPs对宿主细胞有很强的细胞毒性。Hoechst染核结果发现实验组出现了凋亡小体,这与STS处理的阳性对照组中细胞变化相一致。但是ECPs处理后的宿主细胞未出现明显的DNA Ladder。【结论】卵形鲳鲹源创伤弧菌对宿主细胞具有明显的毒性,再通过理化性质的检测,可以初步确定创伤弧菌导致卵形鲳鲹细胞死亡是通过凋亡途径发生的。未来将针对凋亡途径的具体通路进行深入研究,为控制病菌的感染提供理论依据。
关键词:  卵形鲳鲹 创伤弧菌 致死机制 细胞凋亡
DOI:
投稿时间:2019-07-04修订日期:2019-09-03
基金项目:
The apoptosis of host cells induced by vibrio vulnificus from Trachinotus ovatus
liumingzhu1, yuqing1, xiaohehe2, WU Siting3, lisiqiao1, wangyibing4, niezhenping1, lipengfei1
(1.Guangxi Academy of Sciences;2.Henan Normal University;3.Guangxi Key Lab for Marine Biotechnology;4.School of Marine Sciences and Biotechnology)
Abstract:
Abstract:【Objective】Members of the genus Vibrio are autochthonous bacterial flora in the natural ecosystem, and quite a few of them are associated with infections in humans and aquatic animals, some of them are lethiferous. However, as aquaculture expands in recent years, lots of fish diseases outbreaks were induced by V. vulnificus. It not only results in serious economic losses, but also potentially threats human health. V. vulnificus has been isolated and identified from Trachinotus ovatus in the previous study. In this study, we focused on the infection mechanism of V. vulnificus on host cells. The aim of this study is to investigate the lethal mechanism of V. vulnificus, and lay the foundation for the further development of anti-fish disease functional products. 【Methods】In this paper, we first measured the protein concentration of the extracellular bacteria products (ECPs) of V. vulnificus. ECPs at different concentrations were applied to determine the cell toxicity by cell morphology observation and cell viability assay. Hoechst and DNA Ladder were further used to determine the lethal mechanism.【Results】ECPs of V. vulnificus were toxic to host cell. Observation result of nuclear staining by Hoechst 33423 showed that, apoptotic bodies emerged in the ECPs-treated host cells, which was consistent with the positive control cells treated with STS. However, the total DNA of ECPs-treated host cells showed no DNA Ladder. 【Conclusion】V. vulnificus from T. ovatus exhibited high toxicity for host cell. According to the apoptotic bodies in ECPs-treated host cells, V. vulnificus might reduce the death of host cells in apoptotic pathway. In the future, we will study the precise mechanisms of apoptotic pathways in V. vulnificus treated host cells, which would provide theoretical basis for the control of bacterial infection..
Key words:  Trachinotus ovatus, Vibrio vulnificus, extracellular bacteria products, pathogenesis, apoptosis

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