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刘明珠,余庆,肖贺贺,吴思婷,朱冬琳,董德信,覃仙玲,陈宪云,王一兵,聂振平,肖俊,李鹏飞.卵形鲳鲹源创伤弧菌导致宿主细胞凋亡的作用机制[J].广西科学院学报,2019,35(3):206-212. [点击复制]
- LIU Mingzhu,YU Qing,XIAO Hehe,WU Siting,ZHU Donglin,DONG Dexin,QIN Xianling,CHEN Xianyun,WANG Yibing,NIE Zhenping,XIAO Jun,LI Pengfei.The Apoptosis of Host Cells Induced by Vibrio vulnificus from Trachinotus ovatus[J].Journal of Guangxi Academy of Sciences,2019,35(3):206-212. [点击复制]
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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 |
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刘明珠1, 余庆1, 肖贺贺2,3, 吴思婷4, 朱冬琳1, 董德信1, 覃仙玲1, 陈宪云1, 王一兵1,5, 聂振平3, 肖俊4, 李鹏飞3,6
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| (1.广西科学院, 广西近海海洋环境科学重点实验室, 广西南宁 530007;2.河南师范大学生命科学学院, 河南新乡 453007;3.广西壮族自治区海洋研究所, 广西海洋生物技术重点实验室, 广西北海 536000;4.广西壮族自治区水产科学研究院, 广西水产遗传育种与健康养殖重点实验室, 广西南宁 530021;5.广西民族大学海洋与生物技术学院, 广西南宁 530006;6.广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007) |
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| 摘要: |
| 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）引起的鱼病不胜枚举，不仅给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。本文通过确定创伤弧菌胞外产物（Extracellular 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 33342染核观察及DNA ladder来进一步确定其致死机制。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 33342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。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 胞外产物 致死机制 细胞凋亡 水产养殖 |
| DOI：10.13657/j.cnki.gxkxyxb.20190903.002 |
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| 基金项目:广西重点研发计划项目（2018AB52003），广西自然科学基金项目（2017GXNSFBA198176），广西科学院基本科研业务费项目（2018YJJ903，2018YJJ902），现代农业产业技术体系专项资金项目（CARS-46）和广西科技人才专项（2018AD19367）资助。 |
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| The Apoptosis of Host Cells Induced by Vibrio vulnificus from Trachinotus ovatus |
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LIU Mingzhu1, YU Qing1, XIAO Hehe2,3, WU Siting4, ZHU Donglin1, DONG Dexin1, QIN Xianling1, CHEN Xianyun1, WANG Yibing1,5, NIE Zhenping3, XIAO Jun4, LI Pengfei3,6
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| (1.Guangxi Key Laboratory of Marine Environmental Science, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;2.College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China;3.Guangxi Key Laboratory for Marine Biotechnology, Guangxi Institate of Oceanography, Beihai, Guangxi, 536000, China;4.Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Breeding, Guangxi Academy of Fishery Science, Nanning, Guangxi, 530021, China;5.School of Marine Sciences and Biotechnology, Guangxi University for Nationalities, Nanning, Guangxi, 530006, China;6.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Nanning, Guangxi, 530007, China) |
| Abstract: |
| Vibrio is a part of the natural flora.As harmful bacteria,it can not only infect common aquatic products,some of them are also pathogenic to the human body.However,with the continuous expansion of aquaculture in recent years,lots of fish diseases outbreaks were induced by Vibrio vulnificus,which not only caused huge losses to the aquaculture industry,but also potentially threated the safety of human life.In this paper,the infection mechanism of V.vulnificus on host cells was studied in order to elucidate the lethal mechanism of V.vulnificus on host cells and provide a theoretical basis for the future development of anti-fish disease functional products.In this paper,by determining the protein concentration of extracellular products (ECPs) of V.vulnificus,the toxicity of V.vulnificus ECPs was determined by light microscopy and cell viability CCK detection based on different protein concentrations.Hoechst nuclear staining observation and DNA ladder were then used to further determine the lethal mechanism.According to the cytotoxicity results,V.vulnificus ECPs of Trachinotus ovatus had strong cytotoxicity to host cells.Observation result of nuclear staining by Hoechst 33342 showed that apoptotic bodies emerged in the experimental group,which was consistent with cell changes in the STS-treated positive control group.However,the host cells treated with ECPs did not show significant DNA ladder.V.vulnificus from T.ovatus exhibited high toxicity to host cell,and by the detection of physicochemical properties,it could be preliminarily determined that V.vulnificus caused the occurrence of oval cell death through the apoptotic pathway.In the future,we will conduct in-depth research on the specific paths of the apoptotic pathway,which can provide a theoretical basis for controlling the infection of pathogens. |
| Key words: Trachinotus ovatus Vibrio vulnificus ECPs pathogenesis apoptosis aquaculture |
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