山羊IL-10蛋白在金黄色葡萄球菌感染免疫反应中调节机制的研究  

Study on the regulation mechanism of goat IL-10 on immune response to Staphylococcus aureus infection

作　　者：郑姚 李娟[1] 王利[1] 魏勇[2] ZHENG Yao;LI Juan;WANG Li;WEI Yong(Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Ministry of Educationand Sichuan Province,Southwest Minzu University,Chengdu 610041,China;Animal Science Academy of Sichuan Province,Chengdu 610041,China)

机构地区：[1]西南民族大学青藏高原动物遗传资源保护与利用教育部和四川省重点实验室,四川成都610041 [2]四川省畜牧科学研究院,四川成都610041

出　　处：《中国预防兽医学报》2020年第11期1167-1171,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：四川省肉羊创新团队防疫岗位(sccxtd-2020-14);农业农村部农业重大技术协同推广计划试点四川省肉羊高效生产配套技术推广应用。

摘　　要：为探究金堂黑山羊白介素-10蛋白(IL-10)在金黄色葡萄球菌感染免疫反应中的调节机制,本研究构建了重组表达质粒pET32a-IL-10,经诱导后表达纯化获得山羊重组IL-10蛋白(rIL-10),SDS-PAGE和western blot鉴定结果显示,山羊IL-10在大肠杆菌中以包涵体形式表达,重组蛋白大小约为27 ku。每组5只通过金黄色葡萄球菌感染小鼠试验研究rIL-10的免疫调节作用,将15只SPF小鼠随机分为A、B和C组,A组为正常对照组,B和C组腹腔注射500μL金黄色葡萄球菌(1×10^8 cfu/mL),感染24 h后C组腹腔注射200μg/kg rIL-10,感染48 h后检测各组小鼠血清学指标和3种炎症因子在其6种组织中的转录水平,结果显示,B组小鼠血清丙二醛MDA含量极显著高于A和C组(p<0.01),总抗氧化能力T-AOC和谷胱甘肽过氧化物GSH-PX酶活性极显著低于A和C组(p<0.01);IL-1β基因转录水平在B组小鼠心、肝、肺和肾中极显著高于A组(p<0.01),在C组小鼠心、肝、脾和肺中低于B组(p<0.01),在脾和肾中低于A组(p<0.01);炎症因子IL-2在B组小鼠肝、脾、肺和小肠中的转录水平高于A组(p<0.05或p<0.01),在C组小鼠肝、脾、肺、肾和小肠中低于B组(p<0.05或p<0.01),在心、脾和肾中显著低于A组(p<0.05);TNF-α基因转录水平在B组小鼠的小肠中高于A组(p<0.05),在C组小鼠心、脾、肺和小肠中低于B组(p<0.05或p<0.01),在心、脾和肺中低于A组(p<0.05或p<0.01)。上述结果表明,山羊rIL-10通过调节血清MDA含量、T-AOC和GSH-PX活性,以及下调炎症因子基因转录水平,改善因金黄色葡萄球菌引起的小鼠免疫因子紊乱。本研究为深入探究山羊IL-10作用及机制提供科学依据。In order to investigate the anti-inflammatory effects of Jintang black goat interleukin-10 on immune response to Staphylococcus aureus infection.In this study,a recombinant expression plasmid pET32a-IL-10 was constructed.Then,the recombinant IL-10 protein was expressed,and identified by SDS-PAGE and western blot.The results showed that goat IL-10 was expressed as inclusion bodies in E.coli and the size of the recombinant protein was about 27 ku.To explore the immune effect of recombinant protein IL-10 through an experiment of Staphylococcus aureus infection in mice,the test were divided into groups A,B,and C.Group A was the control group.Groups B and C were injected intraperitoneally with 500μL Staphylococcus aureus(1×108 cfu/mL).After 24 hours,group C was injected intraperitoneally with 200μg/kg of recombinant IL-10 protein.48 hours after infection,the serological indicators of each group of mice and the transcription levels of three inflammatory factors in their six murine tissues were detected.The results of the mouse test showed that the serum MDA content in the serum of group B mice was significantly higher than that of groups A and C(p<0.01).The total antioxidant capacity and glutathione peroxidase activity in group B were extremely significantly lower than those in groups A and C(p<0.01).The transcription level of IL-1βin the heart,liver,lung and kidney of group B was significantly higher than that of group A(p<0.01),and that of group C was lower than group B in the heart,liver,spleen and lung(p<0.01).The IL-1βtranscription level of group C was lower than group A,in spleen and kidney(p<0.01).The inflammatory factor IL-2 transcription level of group B was higher than group A in the liver,spleen,lung and small intestine(p<0.05 or p<0.01).The IL-2 transcription level of group C was lower than group B in the liver,spleen,lung,kidney and small intestine(p<0.05 or p<0.01),and significantly lower than A group in the heart,spleen and kidney(p<0.05).The TNF-αtranscription level was higher in the small intesti

关 键 词：白介素-10 原核表达 金黄色葡萄球菌 炎症因子 山羊 

分 类 号：S852.61[农业科学—基础兽医学]