| 黑嘴病病原菌人工感染对中间球海胆吞噬作用相关免疫指标的影响 |
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| 引用本文： | 王中,张伟杰,刘雷,冷晓飞,高洪涛,江会杰,常亚青.黑嘴病病原菌人工感染对中间球海胆吞噬作用相关免疫指标的影响[J].大连海洋大学学报,2021,36(2):241-247. |
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| 作者姓名： | 王中 张伟杰 刘雷 冷晓飞 高洪涛 江会杰 常亚青 |
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| 作者单位： | 大连海洋大学 农业农村部北方海水增养殖重点实验室,辽宁 大连116023;大连海宝渔业有限公司,辽宁 大连116045 |
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| 基金项目： | 农业农村部农业科研杰出人才及创新团队项目;国家重点研发计划项目;国家自然科学基金 |
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| 摘 要： | 为探讨黑嘴病病原菌感染对中间球海胆Strongylocentrotus intermedius吞噬作用相关免疫指标的影响,选用壳径约2 cm的中间球海胆108只,用注射器针头刺入海胆体腔,再用浓度102 CFU/mL的黑嘴病病原菌液浸泡,经病原菌胁迫后0、1、6、12、24、48 h时测定各体腔细胞密度、细胞凋亡率、细胞坏死率、吞噬作用相关免疫指标(酸性磷酸酶(ACP)活性、活性氧(ROS)含量和总抗氧化能力(T-AOC))及吞噬相关免疫基因表达量,并利用有效吞噬细胞比例计算几种免疫指标的单细胞贡献值。结果表明:中间球海胆体腔液中变形吞噬细胞密度在黑嘴病病原菌胁迫1 h后下降约80%,同时吞噬细胞凋亡率上升至60.92%;体腔细胞中ACP、ROS和T-AOC在胁迫后呈现不同程度的变化趋势,计算为单个吞噬细胞平均贡献值后,这些指标及C3-pre和Clec4g基因相对表达量均一致呈现先升高后下降趋势,且均在胁迫1 h时达到最高,分别较胁迫前升高3、6、7、4、7倍;在胁迫后6~24 h,虽然吞噬细胞密度逐渐恢复,但其凋亡率呈下降趋势,坏死率逐渐上升至63.98%,C3-pre和Clec4g基因相对表达量,以及ACP、ROS和T-AOC的单个吞噬细胞平均贡献值均出现下降趋势;在胁迫后48 h,Caspase-8基因相对表达量上调至最高值,约为0 h的2倍。研究表明,中间球海胆在病原菌入侵前期,通过提高ACP活性、ROS含量和T-AOC及吞噬相关免疫基因表达等方式增强吞噬作用,并通过细胞凋亡和再生保持细胞数量,在病原菌入侵后期,由于不能清除病原菌,上述指标均下降,吞噬作用逐渐减退,细胞坏死率大幅上升,导致海胆发病。
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| 关 键 词： | 中间球海胆 黑嘴病 吞噬细胞 免疫指标 基因表达 |
Effects of artificial challenge of black mouth disease pathogen on phagocytosis related immune parameters in sea urchin Strongylocentrotus intermedius |
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| Authors: | WANG Zhong ZHANG Weijie LIU Lei LENG Xiaofei GAO Hongtao JIANG Huijie CHANG Yaqing |
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| Institution: | (Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture and Rural Affairs, Dalian Ocean University, Dalian 116023, China;Dalian Haibao Fisheries Limited Company, Dalian 116045, China) |
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| Abstract: | In order to investigate the effect of black mouth disease pathogen infection on phagocytosis of sea urchin Strongylocentrotus intermedius,the coelomocytes density,apoptosis rate,cell necrosis rate and phagocytosis related immune indices,including acid phosphatase activity(ACP),reactive oxygen species(ROS),total antioxidant capacity(T-AOC)and phagocytosis related immune gene expression were determined 0,1,6,12,24 and 48 h post pathogen stress in the sea urchin damaged by piercing the mouth membrane with a needle into the body cavity and then exposed to black mouth disease pathogen fluid(102 CFU/mL).The single phagocyte contribution value of several immune parameters was calculated by using the proportion of effective phagocyte.The density of phagocytes in coelomic fluid was shown to be decreased by 80%,and the apoptosis rate of phagocytes to be increased to 60.92%1 h post stress.There were different relative trends of ACP activity,ROS level and TAC in coelomocytes after stress.However,the average contribution of single phagocyte showed a consistent trend of first increase and then decrease,with the maximal expression level of C3-pre and Clec4g genes at 1 h post stress,increased by 3 times,6 times,7 times,4 times and 7 times,respectively.The apoptosis rate showed a downward trend,and the necrosis rate gradually was increased to 63.98%.C3-pre and Clec4g genes,as well as the single phagocytemean relative expression of ACP,ROS and T-AOC was decreased to the level before stress from 6 h to 24 h post stress,although the density of phagocytes gradually recovered.The maximal expression level of Caspase-8,as about 2 times as that at 0 h stress,was observed at 48 h post stress.It was found that the phagocytosis of the sea urchin was enhanced by the increase in ACP activity,ROS,T-AOC,and up regulation of phagocytic related immune genes at the beginning of infection.Meanwhile up-regulation of apoptosis rate of phagocytes and cell regeneration were also found to maintain the number of phagocytes.At the late stage of the invasion of pathogens,phagocytes failed to clear all the pathogen,the above phagocytosis related indices gradually were decreased,and the necrosis rate of phagocytes so significantly increased that the sea urchin became diseased. |
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| Keywords: | Strongylocentrotus intermedius black mouth disease phagocyte immune parameter gene expression |
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