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目的筛选及鉴定支气管哮喘(简称哮喘)患者记忆 CD_4~+T 淋巴细胞活化相关基因。方法使用差异显示聚合酶链反应(DDPCR)方法筛选哮喘患者和健康人记忆 CD_4~+T 淋巴细胞差异表达基因,采用逆转录-聚合酶链反应(RT-PCR)法检测差异表达基因的 mRNA 表达水平。统计学处理采用 SPSS 10.0软件。计量资料采用±s表示,组间比较采用双因素方差分析,P<0.05为差异有统计学意义。结果经 DDPCR 克隆获得了19条差异片段,经同源性分析显示其中2条片段分别与白细胞介素-7(IL-7)和 MM-1基因高度同源,采用 RT-PCR 检测进一步证实了这2个基因在哮喘患者激活的记忆 CD_4~+T 淋巴细胞中表达白细胞介素-7和 MMI 基因激活后分别为0.390±0.029、0.629±0.047(F 值分别为983、1264,P 均<0.05)。结论哮喘患者激活的记忆 CD_4~+T 淋巴细胞中 IL-7和MM-1基因的上调表达可能是哮喘患者与健康人对于变应原出现不同反应的分子机制之一。
Objective To screen and identify memory related CD_4 ~ + T lymphocyte activation genes in patients with bronchial asthma (asthma). Methods Differential display polymerase chain reaction (DDPCR) was used to screen differentially expressed CD_4 ~ + T lymphocytes in asthmatic patients and healthy people, and RT-PCR was used to detect mRNA expression of differentially expressed genes Level. Statistical analysis using SPSS 10.0 software. Measurement data using ± s that between groups using two-factor analysis of variance, P <0.05 for the difference was statistically significant. Results A total of 19 differential fragments were obtained by DDPCR cloning. Two of the fragments were highly homologous to IL-7 and MM-1 genes by homology analysis. The results of RT-PCR confirmed that The expressions of IL-7 and MMI in memory CD_4 ~ + T lymphocytes activated by asthma were 0.390 ± 0.029,0.629 ± 0.047 (F = 983,1264, P <0.05 respectively) ). Conclusion The up-regulated expression of IL-7 and MM-1 in activated memory CD_4 ~ + T lymphocytes of asthmatic patients may be one of the molecular mechanisms that alleviates allergens in asthmatic patients and healthy individuals.