结核分枝杆菌抗原对人单核细胞核苷酸结合寡聚化结构域样受体蛋白12表达的影响  

M. tuberculosis antigens inhibit the NLRP12 expression in monocytes from patients with active pulmonary TB

作　　者：刘艳华 王若 程小星 

机构地区：[1]解放军第三0九医院全军结核病研究所全军结核病防治重点实验室结核病诊疗新技术北京市重点实验室,北京100091

出　　处：《中国防痨杂志》2015年第4期339-343,共5页Chinese Journal of Antituberculosis

基　　金：国家自然科学基金青年基金项目（81101219）

摘　　要：目的 研究结核分枝杆菌（Mycobacterium tuberculosis, Mtb）抗原对人单核细胞中核苷酸结合寡聚化结构域样受体蛋白12（NLRP12）表达的影响，为深入研究NLRP12在结核病中的作用奠定实验基础。 方法 采集15例活动性肺结核患者和15名健康人的抗凝血，分离外周血中单核细胞，用Mtb抗原H37Rv全菌裂解液、早期分泌靶抗原6（EAST-6）和培养分泌蛋白10（CFP-10）的抗原多肽刺激，提取单核细胞的总RNA，采用荧光定量PCR方法检测NLRP12 mRNA的表达，采用配对t检验分析刺激前后单核细胞NLRP12的表达差异。以P〈0.05为差异有统计学意义。结果 全菌裂解液、EAST-6和CFP-10抗原多肽刺激后的活动性肺结核患者单核细胞与未刺激前的单核细胞相比，其NLRP12 mRNA的表达显著下调，由未刺激前的（1.8±1.26）×10^-2分别下降到（1.09±0.78）×10^-2（t=3.826，P〈0.01）和（1.14±0.87）×10^-2（t=3.695，P〈0.01）。全菌裂解液、EAST-6和CFP-10抗原多肽刺激后，健康人单核细胞中NLRP12 mRNA的表达与未刺激前相比也降低，由未刺激前的（1.65±0.99）×10^-2分别降至（1.2±0.87）×10^-2（t=1.909，P〉0.05）和（1.38±1.02）×10^-2（t=1.151，P〉0.05），差异无统计学意义。结论 Mtb抗原体外刺激能够抑制活动性肺结核患者单核细胞NLRP12的表达。Objective To study whether Mtb antigens have effect on the NLRP12 expression in peripheral blood monocytes, and provide experimental data to investigate the role of NLRP12 in tuberculosis. Methods Peripheral blood were collected from 15 healthy controls and 15 patients with active pulmonary tuberculosis. Monocytes were isolated from peripheral blood and then stimulated with Mtb antigens, including whole cell lysate of H37Rv strain and ESAT-6/CFP-10 peptide pool. Total RNA was extracted from unstimulated and stimulated cells, NLRP12 mRNA was detected by fluorescence quantitative PCR, and the difference of NLRP12 mRNA expression between stimulated and unstimulated cells was analyzed by paired t test.P value less than 0.05 was considered statistically significant. Results The reduction of NLRP12 mRNA in monocytes from active pulmonary TB was significant following stimulation by whole cell lysate and peptide pool, which reduced to （1.09±0.78）×10^-2 （t=3.826, P〈0.01） and （1.14±0.87）×10^-2 （t=3.695, P〈0.01） from （1.8±1.26）×10^-2, respectively. The expression of NLRP12 mRNA in monocytes from healthy subjects was no significant change following stimulation by whole cell lysate and peptide pool, although which reduced to （1.2±0.87）×10^-2 （t=1.909, P〉0.05） and （1.38±1.02）×10^-2 （t=1.151, P〉0.05） from （1.65±0.99）×10^-2, respectively. Conclusion Mtb antigens can inhibit the NLRP12 expression in monocytes from patients with active pulmonary Tin vitro.

关 键 词：结核 肺 结核分枝杆菌 细胞内信号肽和蛋白质类 单核细胞 重组融合蛋白质类 

分 类 号：R392-33[医药卫生—免疫学]