支气管哮喘不同发作状态时人类白细胞抗原DPB1基因mRNA表达量的变化研究

The study on humon leukocyte antigen-DPB1 gene mRNA expression in different asthma states by FQ-RT-PCR

目的应用实时荧光定量RT-PCR方法检测支气管哮喘急性发作期患者和缓解期患者人类白细胞抗原-DPB1(human leukocyte antigen-DPB1,HLA-DPB1)基因mRNA定量表达情况,探讨HLA-DPB1基因的表达量与支气管哮喘发作状态之间的相关性。方法抽取43例支气管哮喘急性发作期患者、17例支气管哮喘缓解期患者以及34例健康对照者外周血各2ml,Trizol试剂提取外周血白细胞总RNA,利用随机引物逆转录总RNA成cDNA,根据HLA-DPB1第二外显子序列设计序列特异性引物,实时荧光定量PCR检测其mRNA的定量表达情况,以人磷酸甘油醛脱氢酶(g1ucera1dehyde phosphatedehydrogenase,GAPDH)基因为内对照,统计分析三组间差异,P<0.05(双侧)表示差异有显著性。结果HLA-DPB1基因mRNA在支气管哮喘急性发作期、支气管哮喘缓解期以及健康对照组定量表达的情况经过单因素方差分析,其模板cDNA量对数值在支气管哮喘急性发作期患者组为4.243±0.957,支气管哮喘缓解期患者组为2.709±0.629,健康对照组为2.295±0.698。支气管哮喘急性发作期患者组HLA-DPB1基因mRNA表达明显高于健康对照组和支气管哮喘缓解期患者组(P<0.01);支气管哮喘缓解期患者组表达量与健康对照组比较无显著性差异(P>0.05)。结论HLA-DPB1基因在支气管哮喘急性发作期患者中表达明显增高,与支气管哮喘患病状态有相关性。

Objective To study the human leukocyte antigen DPBI（HLA-DPB1） mRNA expression level in acute asthmatics and released asthmatics patients using fluorogenic quantitative reserve transcription polymerize chain reaction （FQ-RT-PCR）, and explore the possible relationship between the HLA-DPB1 mRNA expression level and different asthma states, Methods Forty-three acute asthmatics patients （AA）, 17 released asthmatics patients （RA） and 34 healthy control subjects （HC） were chosen in this experiment, and 2 ml anticoagulated peripheral whole blood was taken from the above three groups, The total RNA was extracted by Trizol reagent in all samples from all the above groups respectively, and the mRNA was transcribed into cDNA using oligo primer. The HLA-DPB1 extron 2 gene was chosen to amplify in vitro with gene specific primer by PQ-RT-PCR. The statistical software SPSS10.0 for Windows was used to compare and analyze the data of the three groups to see if there were any statistical differences. Results The logarithmic value of the cDNA in acute asthmatics group was 4.343±0.957;that of the released asthmatic group was 2.709±0.69; that of the asthmatics group was 2.295±0.698. There was a significant difference in the HLA DPB1 mRNA expression level between acute asthmatic group and healthy control group （P〈0.001）,but in released asthmatic group there was no significant change compared with that of healthy control group （P〉0.05）. Conclusions The expression of HLA-DPB1 mRNA is higher in acute asthmatics patients than in healthy controls, but not in released asthmatics patients, There was a positive relationship between the asthma attacking and the HLA-DPB1 gene expression.