DON、ZEA及其联合作用对小鼠T淋巴细胞活化及MAPK信号通路的影响  

Effects of DON, ZEA and their combination on T lymphocyte activation and MAPK signaling pathway in mice

作　　者：王珺 孙凯[1] 郑豪 邹辉[1] 袁燕[1] 顾建红[1] 刘学忠[1] 刘宗平[1,2] 卞建春 WANG Jun;SUN Kai;ZHENG Hao;ZOU Huil;YUAN Yan;GU Jianhong;LIU Xuezhong;LIU Zongping;BIAN Jianchun(College of Veterinary Medicine,Yangzhou Uni versity,Yangzhou 225009,China;Jiangsu Co-Imnovation Center for Prevention and Control of Im portant Animal In fectious Diseases and Zoonoses,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2020年第1期7-15,共9页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家重点研发计划项目(2016YFD0501208);扬州大学大学生学术科技创新基金项目(x20180683、x20180943);江苏高校优势学科建设工程项目(PAPD)。

摘　　要：为揭示脱氧雪腐镰刀菌烯醇(DON)、玉米赤霉烯酮(ZEA)及其联合作用的免疫抑制机制,试验研究霉菌毒素DON、ZEA单独及联合染毒对小鼠离体脾脏淋巴细胞活性、T淋巴细胞活化及其MAPK信号通路的影响。以小鼠原代混合淋巴细胞为材料,以刀豆蛋白A(Con A)为混合淋巴细胞活化刺激剂,设ZEA、DON单独及联合染毒组,CCK-8法检测不同毒素组处理细胞24 h后细胞活性的变化;流式细胞术检测T淋巴细胞活化标识分子CD25以及共刺激分子CD278(ICOS)的表达情况;Western blot检测T淋巴细胞内MAPK通路相关蛋白ERK、JNK、p38等表达情况。结果表明:与细胞对照组相比,Con A组混合淋巴细胞相对存活率极显著上升;与Con A组相比,ZEA或DON单染组及联合染毒组细胞相对存活率显著或极显著下降。与对照组相比,Con A组CD4^+CD25^+/CD4^+及CD4^+CD278^+/CD4^+比例均极显著上升;与Con A组相比,ZEA、DON及联合染毒组CD4^+CD25^+/CD4^+比例均极显著下降,CD4^+CD278^+/CD4^+比例均不同程度下降,染毒浓度越高下降越明显。此外,不同浓度毒素处理组对T淋巴细胞MAPK通路相关蛋白ERK、JNK、p38等表达均有促进作用。这一研究提示,DON、ZEA及其联合可通过抑制CD25、CD278(ICOS)的表达来干扰T淋巴细胞的活化过程,DON、ZEA及其联合作用通过对MAPK信号通路的影响调控T淋巴细胞的存活。In order to reveal the immunosuppressive mechanism of deoxynivalenol(DON), zearalenone(ZEA) and their combined effects, the effects of mycotoxin DON and ZEA on isolated spleen lymphocytes of mice, T lymphocytes activation and MAPK signaling pathway were studied. In this experiment, mixed primary lymphocytes of mice were used as model. Con A was used as a mixed lymphocyte activation stimulant,setting up ZEA and DON groups separately and jointly. CCK-8 method was used to detect the changes of cell activity after 24 hours treatment in different toxin groups. Flow cytometry was used to detect the expression of CD25, a marker of T lymphocytes activation, and CD278(ICOS), a costimulatory molecule. Western blot was used to detect MAPK pathway-related proteins in T lymphocytes(e.g. The expression of ERK, JNK, p38, etc). The results showed that compared with the control group, the relative survival rate of mixed lymphocytes in Con A group increased significantly(P<0.01);compared with Con A group, the relative survival rate of T lymphocytes in ZEA or DON single exposure group and combined exposure group decreased significantly or very significantly(P<0.05 or P<0.01);compared with the control group, the proportion of CD4^+CD25^+/CD4^+and CD4^+CD278^+/CD4^+in Con A treatment group increased significantly,compared with Con A group,the proportion of CD4^+CD25^+/CD4^+in ZEA,DON and combined exposure group decreased significantly(P<0.01),and the proportion of CD4^+CD278^+/CD4^+in ZEA,DON and combined exposure group decreased to varying degrees,and the higher the concentration of exposure,the more obvious the decrease was.In addition,different toxin treatments promoted the expression of MAPK pathway-related proteins(such as ERK,JNK,p38,etc.)in T lymphocytes.This study shows that DON,ZEA and their combination can interfere with the activation of T lymphocytes by inhibiting the expression of CD25 and CD278(ICOS).In addition,DON,ZEA,and their combination regulate T lymphocytes survival by affecting the MAPK signaling pathway.

关 键 词：玉米赤霉烯酮 脱氧雪腐镰刀菌烯醇 T淋巴细胞活化 CD25 CD278(ICOS) MAPK 

分 类 号：S852.4[农业科学—基础兽医学]