目的对基因表达谱芯片筛选出的重症比对轻症继发性肺结核患者基因CCL3、CCL4、CXCL2进行验证,探索其表达下调与重症继发性肺结核免疫病理机制的关系。方法用Affymetrix基因表达谱芯片检测重症和轻症继发性肺结核患者以及与健康对照相比的差异表达基因;用real-time PCR法测定重症、轻症患者和健康对照CCL3、CCL4、CX-CL2的相对表达量;用方差分析和非参数检验统计方法判断组间比较的统计学意义。结果表达谱芯片筛选出重症继发性肺结核患者有大量差异表达基因,一些免疫基因表现为下调,可能与病变严重程度有相关性。real-time PCR结果显示CCL3在重症vs轻症表达下调(P<0.05),重症vs健康有下调趋势;CCL4在组间比较虽无统计学差异但重症vs轻症、重症vs健康有下调表达趋势;CXCL2在重症vs轻症有下调趋势,相对于健康人有轻度上调(P<0.05)。结论基因表达谱芯片检测CCL3、CCL4、CXCL2在重症vs轻症继发性肺结核患者表达下调,验证结果与芯片基本相符,趋化因子下调可能在重症继发性肺结核免疫病理机制中有重要作用。 Objective To verify the expression of genes CCL3, CCL4 and CXCL2 in critically ill patients with mild secondary pulmonary tuberculosis by gene expression profiling and to explore the relationship between the expression of CCL3 and CXCL2 and the pathological mechanism of severe secondary pulmonary tuberculosis. Methods Affymetrix gene expression microarrays were used to detect differentially expressed genes in patients with severe and mild secondary pulmonary tuberculosis and those in healthy controls. The levels of CCL3, CCL4 and CX-CL2 in severe, mild and healthy controls were determined by real-time PCR Of the relative expression levels; analysis of variance and non-parametric test statistical methods to determine the statistical significance between groups. Results There were a large number of differentially expressed genes in patients with severe secondary pulmonary tuberculosis after expression profiling. Some of the immune genes were downregulated, which may be related to the severity of the disease. The results of real-time PCR showed that CCL3 was down-regulated in critically ill patients and mild ones (P <0.05), while those in severe cases vs healthy ones were down-regulated. CCL4 showed no significant difference between groups (P <0.05). CXCL2 showed a downward trend in critically ill patients and mild patients, with a slight increase (P <0.05). Conclusion The gene expression microarray detection of CCL3, CCL4 and CXCL2 is down-regulated in patients with severe and mild secondary pulmonary tuberculosis. The validation results are consistent with that of the chip. The down-regulation of chemokines may play an important role in the immunopathogenesis of severe secondary tuberculosis.