民猪源唾液联合乳杆菌B112株调控内质网应激预防炎症反应的作用机制  

Mechanism of regulation of plasmic reticulum stress and prevention of inflammation in combination with Ligilactobacillus salivarius B112 from Min pigs

作　　者：吴佳鑫 杨晓钰 李卓然 项晴 刘郁蕾 王晓娜 李佳璇[1,2] 姜艳平 崔文[1,2] 李一经 唐丽杰[1,2] WU Jiaxin;YANG Xiaoyu;LI Zhuoran;XIANG Qing;LIU Yulei;LI Jiaxuan;JIANG Yanping;CUI Wen;LI Yijing;TANG Lijie(College of Veterinary Medicines,Northeast Agricultural University,Harbin 150030,China;Northeast Scientific Inspection Observation Station,Key Laboratory of Animal Pathogen Biology of Ministry of Agriculture,Harbin 150030,China)

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]农业部动物疫病病原生物学重点实验室/东北科学观测实验站,哈尔滨150030

出　　处：《中国兽医科学》2024年第6期784-792,共9页Chinese Veterinary Science

基　　金：国家自然科学基金联合基金项目U22A20527。

摘　　要：本研究旨在探讨民猪源唾液联合乳杆菌(Ligilactobacillus salivarius)通过调控内质网应激预防LPS刺激猪小肠上皮细胞(IPEC-J2)引起炎症的作用机制。本试验从抗病性和耐寒性较好的优良猪种民猪的肠道中分离得到一株菌株B112确定为唾液联合乳杆菌。应用单层平板打孔法测定该菌株和三元猪源唾液联合乳杆菌对大肠杆菌、沙门菌和金黄色葡萄球菌的抑菌圈直径大小,以此来反映菌株的抑菌能力。采用体外细胞模型构建的方法,使用唾液联合乳杆菌B112预处理IPEC-J2,然后用LPS刺激IPEC-J2模拟肠道炎症,并通过实时荧光定量PCR检测炎症相关因子和内质网应激相关因子的变化情况。抑菌试验结果表明,民猪源唾液联合乳杆菌B112对3种致病菌均有抑菌作用,对金黄色葡萄球菌和大肠杆菌的抑菌效果较好;三元猪源唾液联合乳杆菌JCM1231对金黄色葡萄球菌并无抑菌作用,但对沙门菌的抑菌效果强于民猪源杆菌。体外抗炎试验结果表明,民猪源唾液联合乳杆菌B112可以降低由LPS刺激引起的IL-6等炎性相关因子和GRP78等内质网应激相关因子的m RNA表达量。综上所述,从民猪肠道内分离出的唾液联合乳杆菌B112可以抑制大肠杆菌、沙门菌和金黄色葡萄球菌的生长繁殖,并且可以通过调控内质网应激预防LPS刺激IPEC-J2引起的炎症反应,本研究有望为后续进一步开发利用唾液联合乳杆菌在预防仔猪炎症方面的应用研究提供理论基础和技术参考。The aim of this study was to investigate the mechanism by which Ligilactobacillus salivarius of porcine origin alleviates LPS-induced inflammation in porcine small intestine epithelial cells(IPEC-J2)by regulating endoplasmic reticulum stress.In this study,a strain B112 was isolated from the intestinal tract of Min pigs with good disease resistance and cold tolerance and identified as Ligilactobacillus salivarius.The diameter of inhibition zone of Ligilactobacillus salivarius from this strain and Ternary hybrid pig against Escherichia coli,Salmonella and Staphylococcus aureus was detected by single plate punching method.In addition,in vitro cell model construction method was used to pretreat IPEC-J2 with Ligilactobacillus salivarius B112,and then stimulate IPEC-J2 with LPS to simulate intestinal inflammation,and real-time fluorescence quantitative PCR was used to detect the changes of inflammatory factors and endoplasmic reticulum stress related factors.The results of bacteriostatic test showed that Ligilactobacillus salivarius B112 from Min pigs had bacteriostatic effect on all three pathogenic bacteria,and had better bacteriostatic effect on Staphylococcus aureus and Escherichia coli,while Ligilactobacillus salivarius JCM1231 from Ternary hybrid pig had no bacteriostatic effect on Staphylococcus aureus,but its bacteriostatic effect on Salmonella was stronger than that of Bacillus from porcine.The results of in vitro anti-inflammatory tests showed that Ligilactobacillus salivarius B112 from Min pigs could decrease the m RNA expression of LPS induced inflammatory factors such as IL-6 and ER stress factors such as GRP78.In summary,Ligilactobacillus salivarius B112isolated from the intestinal tract of Min pigs can inhibit the growth and reproduction of Escherichia coli,Salmonella and Staphylococcus aureus,and can regulate endoplasmic reticulum stress to relieve the inflammation caused by IPEC-J2 stimulated by LPS.It is expected to provide theoretical basis and technical reference for further development and applicat

关 键 词：民猪 唾液联合乳杆菌 分离鉴定 内质网应激 肠道炎症 

分 类 号：S852.612[农业科学—基础兽医学]