中文摘要：

目的探讨肺结核患者不同中医证型与CD14＋单核细胞白介素-1β（interleukine-1β,IL-1β）、白介素-1受体拮抗剂（interleukine-1 receptor anlagenist,IL-1Ra）表达水平的相关性。方法选取104例肺结核确诊患者为研究对象,其中肺阴亏虚33例,阴虚火旺36例,气阴两虚35例。采集抗凝血并利用密度梯度离心法分离外周血单个核细胞,与CD14＋免疫磁珠孵育后获得单核细胞,使用结核杆菌裂解物刺激单核细胞,同时设立空白对照。12小时后收集细胞上清通过酶联免疫吸附法测定IL-1β、IL-1Ra水平,同时裂解细胞提取总RNA,利用实时定量PCR技术测定IL1B、IL1RN基因相对表达量。通过方差分析比较不同证型肺结核患者IL-1β、IL-1Ra表达水平的差异。结果（1）成功建立检测IL1B、IL1RN基因的实时定量PCR技术,靶基因可以被有效扩增,并且引物具有良好特异性;（2）结核菌刺激后IL-1β蛋白表达水平显著升高,其中肺阴亏虚组显著高于阴虚火旺组和气阴两虚组（P〈0.05）,阴虚火旺组IL-1β分泌水平又显著高于气阴两虚组（P〈0.05）。IL-1B mRNA表达趋势与蛋白表达趋势一致,肺阴亏虚、阴虚火旺、气阴两虚三组中IL-1B基因表达水平依次降低,组与组之间差异均有统计学意义（P〈0.05、P〈0.01）;（3）结核菌刺激能够显著增加IL-1Ra分泌水平,气阴两虚组显著高于其它两组（P〈0.05）,而肺阴亏虚组和阴虚火旺组差异无统计学意义（P〉0.05）。同时,在mRNA层面气阴两虚组IL1RN表达水平显著高于其它两组,差异有统计学意义（P〈0.05、P〈0.01）;（4）在结核菌刺激后,IL-1β/IL-1Ra比值在肺阴亏虚和阴虚火旺组中明显增高,显著高于气阴两虚组（P〈0.01、P〈0.05）。结论 IL-1β、IL-1Ra表达水平与肺结核中医病机演变密切相关,由此推测它们不仅可以作为肺结核中医微观辨证的生物标识,也可以作为未来结核病治疗的有效分子靶

英文摘要：

Objective To explore the discrepancy between IL-1β and IL-1Ra expression in CD14＋ monocytes of different TCM syndrome types of puhnonary tuberculosis patients. Methods Totally 104 pulmonary tuberculosis patients were recruited, the patients were divided into Lung-Yin deficiency（ n = 33） ,Yin-Deficiency and Fire-Hyperactivity （ n = 36）, and Qi and Yin deficiency （ n = 35 ）. The density gradient centrifugation was used to separate PBMCs from peripheral blood, then CD14 ＋ monocytes were purified after incubation with magnetic beads. Monocytes were stimulated with or without Mtb lysate for 12 hours,the supernatant was collected to detect IL-1 β and IL-1Ra levels using ELISA, and total RNA was purified from cells and gene expression of IL1B and ILl RN was determined with real-time qPCR assay. Finally,the discrepancy was compared with ANOVA analysis. Results （ 1 ） The expression of ILl B and ILl RN could be detected by real-time quantitative PCR assay with gene specific primers. （2） After Mtb stimulation, IL-1 βlevel was the highest in Lung-Yin deficiency group compared to Yin-Deficiency and Fire- Hyperactivity group and Qi and Yin deficiency group （P〈 0. 05, P〈 0. 01 ）, and it was higher in Yin- Deficiency and Fire-Hyperactivity group than that in Qi and Yin deficiency group（ P〈0.05 ）. There was similar pattern observed in IL1B mRNA level, and IL1B level decreased in the order of Lung-Yin deficiency, Yin-Deficiency and Fire-Hyperactivity, and Qi and Yin deficiency （ P〈0. 05 ,0. 01 ）. （ 3 ） IL-1 Ra expression increased significantly after Mtb stimulation, it was the highest in Qi and Yin deficiency group compared to other two groups （P〈0. 05）, and there was no difference between Yin-Deficicncy and Fire-Hyperactivity group and Qi and Yin deficiency group（ P〉0. 05 ）. Similar pattern was also observed in IL1RN level,which Qi and Yin deficiency group had the highest level（ P〈0. 05, P〈0. 01 ）. （4） After Mtb stimulation, IL-1β/IL-1Ra ratio increased