重症继发性肺结核患者差异表达基因和IL-1β表达的验证

Differentially expressed gene and IL-1β expression in severe secondary pulmonary tuberculosis patients

目的对基因表达谱芯片中筛选的差异表达基因IL-1β进行验证,分析其表达下调与重症继发性肺结核免疫病理的关系。方法 (1)用Affymetrix基因表达谱芯片对重症继发性肺结核(重症)、轻症继发性肺结核(轻症)患者和正常对照者(对照)筛选差异表达基因;(2)用实时荧光定量PCR(RT-PCR)扩增来测定重症、轻症患者和对照IL-1β相对表达量;(3)ELISA法测定IL-1β在重症、轻症患者和对照血浆中的蛋白表达量;(4)用方差分析和非参数检验统计方法判断组间比较的差异,P<0.05为差异有统计学意义。结果 (1)芯片筛选差异基因显示重症肺结核患者IL-1β表达下调13.06倍,参与30个信号通路;(2)RT-PCR结果显示IL-1β在重症和对照组表达下调,P<0.05;(3)ELISA验证IL-1β在重症及轻症组和重症及对照组均表达下调,P<0.05,与芯片结果相符。结论重症继发性肺结核患者IL-1β表达下调,IL-1β可能是其肺结构免疫病理损伤发生发展的重要因素之一。

ObjectiveTo investigate the relationship between downregulation of IL-1β and immunopathology of severe secondary TB with the gene chip selection results. Methods Differentially expressed genes were screened with Affymetrix Gene expression chips for severe and mild secondary TB patients, and healthy controls （they were severe group and mild group and healthy controls respectively）. The relative transcript level of IL-1β was examined by real time PCR （RT-PCR）. The protein level of IL-1βwas detected by ELISA. ANOVA and non-parametric tests were used for statistic analysis among the groups.Results The result of gene chip analysis showed that IL-1β wasreduced 13.06 times in severe group compared with mild group patients. IL-1β was involved in 30 signaling pathways. The result of RT-PCR verified that IL-1β wasreduced in severe group versus healthy controls （P〈0.05）. The result of ELISA verified that IL-1β wasreduced in severe group versus mild group and in severe group versus healthy controls （P〈0.05）. Conclusion IL-1β wasreduced in severe secondary TB. It may involve in immunopathology mechanism for lung tissue damages.