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刘明珠,余庆,肖贺贺,黎思巧,柯珂,覃仙玲,陈宪云,朱冬琳,许尤厚,李鹏飞.维氏气单胞菌对草鱼背鳍细胞的毒性及凋亡研究[J].广西科学院学报,2019,35(3):213-218. [点击复制]
- LIU Mingzhu,YU Qing,XIAO Hehe,LI Siqiao,KE Ke,QIN Xianling,CHEN Xianyun,ZHU Donglin,XU Youhou,LI Pengfei.Study on Cytotoxicity and Apoptosis of Aeromonas veronii to Grass Carp Pectoral Fin Cell[J].Journal of Guangxi Academy of Sciences,2019,35(3):213-218. [点击复制]
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| 维氏气单胞菌对草鱼背鳍细胞的毒性及凋亡研究 |
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刘明珠1, 余庆2, 肖贺贺3, 黎思巧4, 柯珂1, 覃仙玲1, 陈宪云1, 朱冬琳1, 许尤厚5, 李鹏飞1,4
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| (1.广西科学院, 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;2.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;3.河南师范大学生命科学学院, 河南新乡 453007;4.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;5.北部湾大学海洋学院, 广西北部湾海洋生物多样性养护重点实验室, 广西钦州 535011) |
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| 摘要: |
| 草鱼（Ctenopharyngodon idellus）在我国是一种重要的淡水经济鱼类，但是由于近些年的集约化高密度养殖导致各种水产疫病暴发，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，维氏气单胞菌（Aeromonas veronii）就是其中一种人畜共患的条件致病菌。本研究旨在探索维氏气单胞菌导致宿主细胞死亡的致死机制，为今后进一步针对维氏气单胞菌开发抗菌渔用功能产品提供数据支持和理论依据。本文首先测定维氏气单胞菌胞外分泌物（Extracellular products，ECPs）的蛋白浓度，然后以不同浓度的ECPs为基准，根据细胞光镜观察结果及细胞活力检测结果确定维氏气单胞菌胞外ECPs的细胞毒性；再利用Hoechst 33342及TdT-mediated dUTP Nick-End Labeling（TUNEL）检测细胞核的变化情况，最终确定草鱼源维氏气单胞菌对宿主细胞的毒性致死机制。从毒性实验中可以看出高浓度的维氏气单胞菌ECPs对草鱼背鳍（Grass carp pectoral fin，GCPF）细胞具有明显的毒性，且从Hoechst 33342结果可以看到凋亡小体的出现，而TUNEL也出现了阳性结果，实验组出现了绿色荧光，而对照组没有出现荧光。说明维氏气单胞菌胞外分泌物对宿主细胞具有明显的毒性，而且会导致宿主细胞发生细胞凋亡。 |
| 关键词: 草鱼 维氏气单胞菌 胞外分泌物 致死机制 细胞凋亡 |
| DOI：10.13657/j.cnki.gxkxyxb.20190903.003 |
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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），广西科技人才专项（2018AD19367），广西科学院基本科研业务费项目（2018YJJ903，2017YJJ23002）和广西科技重大专项（AA17204095-10）资助。 |
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| Study on Cytotoxicity and Apoptosis of Aeromonas veronii to Grass Carp Pectoral Fin Cell |
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LIU Mingzhu1, YU Qing2, XIAO Hehe3, LI Siqiao4, KE Ke1, QIN Xianling1, CHEN Xianyun1, ZHU Donglin1, XU Youhou5, LI Pengfei1,4
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| (1.Guangxi Key Laboratory for Marine Biotechnology, Guangxi Institute of Oceanography, Guangxi Academy of Sciences, Beihai, Guangxi, 536000, China;2.Guangxi Key Laboratory of Marine Environmental Science, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;3.College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China;4.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Nanning, Guangxi, 530007, China;5.Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, College of Marine Sciences, Beibu Gulf University, Qinzhou, Guangxi, 535011, China) |
| Abstract: |
| Grass carp (Ctenopharyngodon idellus) is an important commercially cultured freshwater fish in China.In recent years,with the advent of intensive aquatic farming,various diseases emerged in commercial populations,which caused enormous economic loss,even threatened human safety.Aeromonas veronii is one of the zoonotic pathogens.This study aimed to explore the lethal mechanism of A.veronii causing to host cell death and provide data supports and theoretical basis for further development of anti-bacterial fishery functional products against A.veronii.This paper first determined the protein concentration of extracellular products (ECPs) of A.veronii.Then,based on the ECPs of different concentrations,the cell toxicity of extracellular ECPs of A.veronii was determined according to the results of light microscopy and cell viability.Hoechst 33342 and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to detect the changes of nucleus,and finally the toxic lethal mechanism of A.veronii to host cells was determined.It can be seen from the toxicity experiment that high concentrations of A.veronii ECPs have obvious toxicity to grass carp pectoral fin (GCPF) cells,and the appearance of apoptotic bodies can be seen from Hoechst 33342 results.Meanwhile,TUNEL showed positive results with green fluorescence in test group and no fluorescence in the control group.It is indicated that A.veronii ECPs is obviously toxic to host cells and can cause apoptosis of host cells. |
| Key words: grass carp Aeromonas veronii ECPs pathogenesis apoptosis |
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