大豆抗原蛋白7S和11S对凡纳滨对虾肠道细胞的致敏机制研究  

Study on Sensitization Mechanism of Soybean Antigen Proteins on Intestinal Cells of Litopenaeus vannamei

作　　者：田仁红 邱建强 彭凯 孙文豪 董芮绮 鲁慧杰 赵红霞 陈冰 陈文淳 郭慧[1] 朱喜锋 李国立 黄文 TIAN Renhong;QIU Jianqiang;PENG Kai;SUN Wenhao;DONG Ruiqi;LU Huijie;ZHAO Hongxia;CHEN Bing;CHEN Wenchun;GUO Hui;ZHU Xifeng;LI Guoli;HUANG Wen(College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,China;Key Laboratory of Animal Nutrition and Feed Science in South China,Ministry of Agriculture in Rural Affairs,Guangdong Key Laboratory of Animal Breeding and Nutrition,Collaborative Innovation Center of Aquatic Sciences,Institute of Animal Science,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China;College of Fisheries and Life,Shanghai Ocean University,Shanghai 201306,China;Guangzhou Fishtech Biotechnology Co.,Ltd.,Guangzhou 510640,China)

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省农业科学院动物科学研究所,广东省农业科学院水产协同创新中心,广东省畜禽育种与营养研究重点实验室,农业农村部华南动物营养与饲料重点实验室,广州510640 [3]华中农业大学水产学院,武汉430070 [4]上海海洋大学水产与生命学院,上海201306 [5]广州飞禧特生物科技有限公司,广州510640

出　　处：《动物营养学报》2025年第1期536-549,共14页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家外国专家项目(G2023030042L);广东省乡村振兴战略专项(2022SPY00007,2022SDZG01);广东省农业科学院水产协同创新中心课题(XT202301)。

摘　　要：本试验旨在探究大豆抗原蛋白7S和11S对凡纳滨对虾肠道细胞的致敏机制。以凡纳滨对虾肠道细胞为研究对象,试验分为对照组(NC组)、7S组、11S组,每组3个重复。细胞培养48 h后,测定各组细胞活性、细胞膜完整性相关指标、细胞抗氧化指标、细胞炎症因子含量、细胞凋亡率、细胞凋亡与紧密连接蛋白相关基因表达。结果表明:与NC组相比,7S组和11S组碱性磷酸酶(AKP)、乳酸脱氢酶(LDH)活性及活性氧(ROS)、丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)含量显著升高(P<0.05),总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及白细胞介素-10(IL-10)含量显著降低(P<0.05),细胞凋亡率显著升高(P<0.05),半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、B淋巴细胞瘤/白血病-2相关X蛋白(Bax)、c-Jun氨基末端激酶(JNK)、p38丝裂原活化蛋白激酶(p38 MAPK)mRNA相对表达量显著上调(P<0.05),紧密连接蛋白-3(Claudin-3)、闭合蛋白(Occludin)和闭锁小带蛋白-1(ZO-1)mRNA相对表达量显著下调(P<0.05)。大豆抗原蛋白7S诱导ROS累积、加剧炎症反应、促进细胞凋亡及破坏细胞屏障的效果强于大豆抗原蛋白11S。综上所述,大豆抗原蛋白7S和11S通过破坏细胞膜完整性,释放AKP和LDH,诱导ROS累积,促进细胞促炎因子分泌,上调细胞凋亡因子和紧密连接蛋白基因表达水平,抑制细胞活性并导致细胞过敏反应。This experiment was conducted to explore the sensitization mechanism of soybean antigen proteins 7S and 11S on intestinal cells of Litopenaeus vannamei.The intestinal cells of Litopenaeus vannamei were divided into control group(NC group),7S group and 11S group with three replicates in each group.After 48 h of cell culture,the cell activity,cell membrane integrity-related indexes,cell antioxidant indexes,cell inflammatory factor contents,apoptosis rate,and apoptosis and tight junction protein-related gene expression were determined in each group.The results showed that compared with NC group,the alkaline phosphatase(AKP),lactate dehydrogenase(LDH)activities and reactive oxygen species(ROS),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)contents of 7S group and 11S group were significantly increased(P<0.05),the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),catalase(CAT)activities and interleukin-10 content were significantly decreased(P<0.05),the apoptosis rate was significantly increased(P<0.05),the mRNA relative expression levels of cysteine aspartate protease-3(Caspase-3),B-lymphoblastoma/leukaemia-2 related X protein(Bax),c-Jun amino-terminal kinase(JNK)and p38 mitogen-activated protein kinase(p 38 MAPK)were significantly up-regulated(P<0.05),and the mRNA relative expression levels of tight junction protein-3(Claudin-3),Occludin and zonula occludens-1(ZO-1)was significantly down-regulated(P<0.05).The soybean antigen protein 7S induced ROS accumulation,exacerbated the inflammatory response,promoted apoptosis and disrupted the cellular barrier more strongly than soybean antigen protein 11S.In summary,soybean antigenic proteins 7S and 11S inhibit cellular activity and lead to cellular anaphylaxis by disrupting cell membrane integrity,releasing AKP and LDH,inducing the accumulation of ROS,promoting cellular pro-inflammatory factor secretion in cells,and up-regulating the expression levels of genes associated with apoptotic factors and tight junction proteins.

关 键 词：凡纳滨对虾 大豆抗原蛋白 肠道细胞 致敏机制 

分 类 号：S963[农业科学—水产养殖]