Caspase-1对小胶质细胞中NOD2介导的免疫耐受的调节  

Caspase-1 activation contributes to Nod2-induced tolerance in microglia cells

作　　者：袁睿莉 孙若鹏[1] 刘心洁[1] 

机构地区：[1]山东大学齐鲁医院小儿内科,山东济南250012

出　　处：《山东大学学报（医学版）》2015年第2期34-38,42,共6页Journal of Shandong University:Health Sciences

基　　金：山东省科技发展计划(2010GSF10262);济南市科学技术局自主创新项目(201004057);国家自然科学基金(81371337)

摘　　要：目的观察小胶质细胞中是否存在核苷酸结合寡聚化结构域蛋白2(NOD2)介导的免疫耐受现象,并探讨其与Caspase-1的关系。方法分别采用不同浓度的胞壁酸二肽(MDP)或肺炎链球菌刺激小胶质细胞24 h,Western blotting检测各组NOD2和核因子κB(NF-κB)的表达情况,ELISA检测白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的表达情况;用MDP(100μg/m L)分别预刺激细胞0、3、24 h后再用同浓度MDP刺激24 h,用ELISA检测IL-6和TNF-α的表达量;Caspase-1活性被ATP激活或被其抑制剂抑制后检测IL-6和TNF-α的变化情况。结果与无刺激处理相比,MDP或肺炎链球菌刺激小胶质细胞24 h后,NOD2、NF-κB、IL-6和TNF-α的表达量均明显增加(P<0.05);MDP分别预刺激细胞0、3、24 h后再刺激24 h,发现预刺激3 h和24 h的细胞IL-6和TNF-α的表达量均低于预刺激0 h;通过ATP激活Caspase-1活性可抑制细胞IL-6和TNF-α的表达,抑制Caspase-1活性可促进IL-6和TNF-α的表达。结论长时间MDP刺激可以诱导小胶质细胞的免疫耐受现象,此现象与Caspase-1的活性具有一定关系。Objective To investigate whether the nucleotide-binding oligomerization-2（NOD2）-mideated tolerance phe-nomenon exists in microglia cells and its relationship with Caspase-1 activity.Methods After microglia cells were stimu-lated with different concentrations of MDP or S.pneumonia for 24 h,the expressions of NOD2 and NF-κB were detected by Western blotting and the expressions of IL-6 and TNF-αwere detected by ELISA.After microglia cells were pre-trea-ted with MDP （100 μg/mL）for 0,3 or 24 h,the expressions of IL-6 and TNF-αwere detected by ELISA to verify the NOD2-mediated tolerance in microglia.After Caspase-1 activity was activated or suppressed by ATP or Caspase-1 inhibi-tor,the expressions of IL-6 and TNF-αwere detected by ELISA to observe the relationship between NOD2-mediated toler-ance and Caspase-1.Results Compared with no treatment,MDP and S.pneumonia treatment for 24 h significantly increased the levels of NOD2,NF-κB,IL-6 and TNF-αin microglia cells（P 〈0.05）.MDP pretreatment for 3 or 24 h reduced secretion of IL-6 and TNF-αupon subsequent stimulation of MDP for 24 h.Levels of IL-6 and TNF-αwere sig-nificantly down-regulated by ATP pretreatment,while up-regulated by Caspase-1 inhibitor treatment.Conclusion Prolonged periods of MDP exposure can induce NOD2-mediated tolerance in microglia cells and NOD2-mediated tolerance is associated with Caspase-1 activity.

关 键 词：小胶质细胞 核苷酸结合寡聚化结构域蛋白 2 耐受 半胱氨酸蛋白酶-1 三磷酸腺苷 

分 类 号：R392.32[医药卫生—免疫学]