三种腺苷受体mRNA表达与哮喘发病及第1秒用力呼气值的相关性

Association of three kinds of adenosine receptors mRNA expression with asthmatic attack and forced expiratory volume in the first second

目的:通过诱导痰技术检测3种腺苷受体mRNA的表达与哮喘发病及第1秒用力呼气值的相关性。方法:实验于2003-10/2004-06在南京大学医学院附属鼓楼医院中心实验室完成。选择该院呼吸科门诊及住院患者9例(哮喘组),女7例,男2例;均无吸烟史,至少1个月内未用过糖皮质激素类药物。选择南京大学医学院研究生和职工9例为正常组,皆为健康正常人,无吸烟及过敏性疾病史。实验经本院伦理委员会批准,并与受试者签定知情同意书。取两组对象的诱导痰;贴壁法分离和纯化巨噬细胞及炎性细胞,并计算相对构成计数的百分比;利用反转录-多聚酶链反应技术和图像分析半定量法检测诱导痰肺泡巨噬细胞腺苷受体mRNA的表达;计算第1秒用力呼气量占预测值百分比与腺苷受体mRNA的相关性。结果:按意向处理分析,18例均进入结果分析。①哮喘组诱导痰肺泡巨噬细胞表达A1及A2b和A3腺苷受体mRNA吸光度积分扫描相对比值均明显高于正常组(0.44±0.11,0.48±0.07,0.48±0.11;0.29±0.06,0.30±0.10,0.36±0.09,t=2.8,4.63,2.85,P<0.05～0.01)。②哮喘组嗜酸性粒细胞、淋巴细胞相对计数构成比明显高于正常组(P<0.01),而巨噬细胞相对计数构成比明显低于正常组(P<0.01)。③第1秒用力呼气量占预测值百分比在哮喘组与A2b腺苷受体mRNA呈正相关(r=0.49,P<0.05)。而与A1及A3腺苷受体mRNA表达无相关性(P>0.05)。结论:A1,A2b,A3腺苷受体可能参与哮喘发病过程,且在哮喘患者诱导痰巨噬细胞与外周血单核细胞表达均高于正常人。但腺苷受体亚型与第1秒用力呼气量占预测值百分比的相关性并不一致,对气道阻塞程度的影响可能不同,有待于进一步证实。

AIM:To investigate the relationship of the expression of three adenosine receptors mRNA with asthmatic attack and forced expiratory volume in the first second by means of induced sputum.METHODS:The experiment was carried out in the central laboratory of Gulou Hospital affiliated to the Medical College of Nanjing University between October 2003 and June 2004.Nine patients with asthma(asthma group,7 females and 2 males),who were hospitalized or treated in the Department of Respiratory Medicine in this hospital,were selected,they all had no smoking history,and had not taken any drugs of glucocorticoid within at least 1 month.Nine healthy postgraduates and employees(control group),who had no history of smoking and allergic disease,were selected from the Medical College of Nanjing University.This experiment obtained the approval from the ethics committee of this college,and all the subjects were involved in this experiment with their permission in written form.The induced sputum samples of the subjects were collected.The macrophages and inflammatory cells were separated and purified with the stick to wall method,and the percentage of relative constitute counting was calculated.The expressions of adenosine receptors mRNA on induced sputum alveolar macrophages were determined with the reverse transcription polymerase chain reaction(RT PCR) and image pattern analysis semi quantitation.The correlation between the percentage of forced expiratory volume in the first second to the predicted value and adenosine receptors mRNA was calculated.RESULTS:According to intention to treat analysis,all the 18 cases were involved in the analysis of results.① The absorbance score scanning relative ratios of adenosine receptors with the expressions of A1,A2b and A3 on induced sputum alveolar macrophages were obviously higher in the asthma group than in the normal group(0.44± 0.11,0.48± 0.07,0.48± 0.11;0.29± 0.06,0.30± 0.10,0.36± 0.09,t=2.8,4.63,2.85,P< 0.05 to 0.01).② The percentages of relative constitute counting of eosinophil and lymphocytes were obviously higher in the asthma group than in the normal group(P< 0.01),but that of macrophage was markedly lower in the asthma group than in the normal group(P< 0.01).③ In the asthma group, the percentage of forced expiratory volume in the first second to the predicted value was positively correlated with A2b adenosine receptors mRNA(r =0.49,P< 0.05),but had no association with A1,A3 adenosine receptors mRNA(P >0.05).CONCLUSION:A1,A2b and A3 adenosine receptors may be involved in the process of asthmatic attack,and the expressions of macrophage in induced sputum and the expression of monocyte in peripheral blood are higher in patients with asthma than in normal people.But the correlation between the subtype of adenosine receptors with the percentage of forced expiratory volume in the first second to the predicted value is not concordant, and the influence on the degree of airway obstruction may be different,which needs further identification.