大肠上皮细胞中MDP活化NOD2诱导耐受性CXCL1、CXCL2、CXCL3和CXCL8表达的分子机制  

The Molecular Mechanism of NOD2 Activated by MDP in Colorectal Epithelial Cells to Induce Tolerant CXCL1,CXCL2,CXCL3 and CXCL8

作　　者：张朝 曹婷[1] 胡玲[1] 胡锦跃 ZHANG Zhao;CAO Ting;HU Ling;HU Jinyue(Department of Clinical Laboratory,the Affiliated Changsha Central Hospital,Hengyang Medical School,University of South China,Changsha 410004,China;Central Laboratory,the Affiliated Changsha Central Hospital,Hengyang Medical School,University of South China,Changsha 410004,China)

机构地区：[1]南华大学附属长沙中心医院检验科,长沙410004 [2]南华大学附属长沙中心医院中心实验室,长沙410004

出　　处：《中国细胞生物学学报》2021年第11期2142-2148,共7页Chinese Journal of Cell Biology

基　　金：湖南省卫生健康委科研计划课题(批准号:2020JJ4636);湖南省自然科学基金(批准号:20201386);南华大学研究生科研创新项目(批准号:203YXC028)资助的课题。

摘　　要：该文探讨肠上皮细胞中胞壁酰二肽(muramyl dipeptide,MDP)活化NOD2诱导趋化因子表达的调控机制。采用RT-PCR和qRT-PCR的方法检测趋化因子、细胞因子及泛素编辑酶A20的mRNA表达水平。Western blot检测A20的蛋白表达水平。ELISA检测趋化因子CXCL8(又称IL-8)的水平。脂质体转染法过表达A20。结果显示,MDP处理HCT116细胞诱导趋化因子CXCL1、CXCL2、CXCL3和CXCL8的表达,但不诱导促炎因子TNFα、IL-1β和IL-6的表达;MDP所诱导的反应具有耐受性,初次处理后的再次处理所诱导的反应程度显著下降;MDP处理上调A20,但过表达A20并不下调MDP诱导的免疫反应;同时,IL-1β上调A20,但IL-1β预处理同样不能下调MDP诱导的反应。总之,大肠上皮细胞中MDP诱导耐受性CXCL1、CXCL2、CXCL3和CXCL8的表达,且A20不参与此耐受机制的形成。This study explored the regulatory mechanism of MDP (muramyl dipeptide) activated NOD2 to induce chemokine expression in colorectal epithelial cells.RT-PCR and qRT-PCR were used to detect mRNA levels of chemokines,cytokines and ubiquitin editing enzyme A20.Western blot was used to detect protein level of A20.ELISA was used to detect protein level of chemokine CXCL8 (also called IL-8).The liposome transfection method was used to over-express A20.The results showed that MDP treatment of HCT116 cells induced the expression of chemokines CXCL1,CXCL2,CXCL3 and CXCL8,but did not induce the expression of pro-inflammatory factors TNFα,IL-1β and IL-6.The production of chemokines induced by MDP was tolerent,because the chemokine levels induced by the re-treatment with MDP after a pre-treatment with MDP was decreased significantly compared with the non-pre-treated group.MDP treatment up-regulated A20,but A20 overexpression did not down-regulate the immune response induced by MDP.Meanwhile,IL-1β up-regulated A20,but IL-1β pre-treatment also failed to down-regulate MDP-induced immune response.In conclusion,MDP induces the tolerant expression of CXCL1,CXCL2,CXCL3 and CXCL8 in colorectal epithelial cells,and A20 does not participate in the formation of this tolerance mechanism.

关 键 词：核苷酸结合寡聚化结构域2 胞壁酰二肽 趋化因子 免疫耐受 大肠上皮细胞 

分 类 号：R392[医药卫生—免疫学]