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[1]张国良 王玲玲 詹森林 曹廷智 汪文斐 张红梅 邓群益 聂广.不同中医证型肺结核患者外周血CD14+单核细胞IL-1β、IL-1Ra表达水平的差异[J].环球中医药,2016,9(10):1170-1174.[doi:10.3969/j.issn.1674-1749.2016.10.003]
　ZHANG Guo-liang,WANG Ling-ling,ZHAN Sen-lin,et al.Discrepancy between IL-1β and IL-1Ra expression in CD14+ monocytes of different TCM syndrome types of pulmonary tuberculosis patients[J].,2016,9(10):1170-1174.[doi:10.3969/j.issn.1674-1749.2016.10.003]
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不同中医证型肺结核患者外周血CD14+单核细胞IL-1β、IL-1Ra表达水平的差异()
     
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《环球中医药》[ISSN:1006-6977/CN:61-1281/TN]

卷:
第9卷
期数:
2016年10期
页码:
1170-1174
栏目:
论著
出版日期:
2016-10-06

文章信息/Info

Title:
Discrepancy between IL-1β and IL-1Ra expression in CD14+ monocytes of different TCM syndrome types of pulmonary tuberculosis patients
作者:
张国良 王玲玲 詹森林 曹廷智 汪文斐 张红梅 邓群益 聂广
518112 深圳市第三人民医院肝病研究所(张国良、詹森林、汪文斐、聂广),结核病科(王玲玲、曹廷智、张红梅、邓群益)
Author(s):
ZHANG Guo-liang; WANG Ling-ling; ZHAN Sen-lin; et al.
Institute of Hepatology,Shenzhen Third People's Hospital,Shenzhen 518112,China.
关键词:
肺结核;  中医证型;  白介素-1β;  白介素-1受体拮抗剂;  单核细胞
Keywords:
Pulmonary tuberculosis;  TCM syndrome types;  Interleukine-1β;  Interleukine-1 receptor antagonist;  Monocytes
分类号:
R249
DOI:
10.3969/j.issn.1674-1749.2016.10.003
文献标志码:
A
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点。
Abstract:
Objective To explore the discrepancy between IL-1β and IL-1Ra expression in CD14+ monocytes of different TCM syndrome types of pulmonary 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-1β and IL-1Ra levels using ELISA,and total RNA was purified from cells and gene expression of IL1B and IL1RN was determined with real-time qPCR assay. Finally,the discrepancy was compared with ANOVA analysis. Results(1)The expression of IL1B and IL1RN could be detected by real-time quantitative PCR assay with gene specific primers.(2)After Mtb stimulation,IL-1β level 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-Deficiency 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 in Lung-Yin deficiency and Yin-Deficiency and Fire-Hyperactivity groups,which was higher than that in Qi and Yin deficiency(P<0.01, P<0.05). Conclusion Expression of IL-1β and IL-1Ra was associated with TCM pathogenesis evolution of pulmonary TB patients,which could be considered as biomarkers of microcosmic syndrome differentiation and effectively therapeutic targets.

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备注/Memo

备注/Memo:
基金项目: 国家自然科学基金(81501714); 广东省自然科学基金(2014A030313789); 深圳市科技计划项目(JCYJ20140411111718166)
作者简介:张国良(1982- ),博士,副主任医师。研究方向:感染病中西医结合诊治研究。E-mail:szdsyy@aliyun.com
更新日期/Last Update: 2016-10-06