氨茶碱对不同密度嗜酸细胞凋亡及其GM-CSF受体αmRNA表达的影响

Influences of aminophylline on expression of GM-CSFRα mRNA and apoptosis in eosinophils in vitro

目的观察氨茶碱(AM)对哮喘豚鼠体外不同密度嗜酸细胞(Eos)凋亡及粒细胞-巨噬细胞克隆剌激因子受体α(GM-CSFRα)mRNA表达的影响, 探讨AM促进哮喘Eos凋亡的机制.方法卵蛋白激发哮喘豚鼠动物模型48h后行支气管肺泡灌洗, 分离低密度及正常密度Eos(HEos,NEos).HEos及NEos分别与AM(10-6～10-3mol/L)共培养24h,以原位杂交方法检测不同Eos的GM-CSFRα mRNA表达, TUNEL法检测细胞凋亡.结果 AM干预体外不同密度Eos24h后,可见Eos细胞凋亡随AM浓度增加而增加,不同密度Eos GM-CSFRα mRNA表达也增加,并与AM浓度呈剂量依赖性.与NEos相比,HEos表达GM-CSFRα mRNA增加明显.HEos表达GM-CSFRα在10-4mol/L 时出现显著差异(P<0.05),而NEos在AM达到最高浓度10-3mol/L 时仍未出现显著差异(P>0.05).GM-CSFRα表达与Eos凋亡呈正相关(P<0.05).结论 AM可促进不同密度Eos表达GM-CSFRα,抑制其细胞因子活动,促进Eos凋亡.AM可通过增加嗜酸细胞GM-CSFRα表达调节Eos凋亡.

Objective To investigate the effects of aminophylline （AM） on expression of GM-CSFRa mRNA and apoptosis in eosinophils（Eos） of bronchalveolar lavage fluid（BALF） in asthmatic guinea pigs, and mechanism of AM promoting Eos apoptosis. Methods After guineas pigs were stimulated by ovalbumin（OVA） for 48h, hypodense Eos （HEos） and normodense Eos （NEos） were purified from BALF by gradients of percoll. Eos cultured in PRMI-1640 medium. Add AM （10^-6-10^-3 mol/L） in the wells. The mRNA of GM-CSFRain Eos was measured by hybridization, apoptosis was detected by TdT-mediated dUTP nick end labeling （TUNEL）. Results Eos cultured under the condition of AM administration in vitro, apoptotic Eos increased, GM-CSFRa mRNA expression increased in a close-dependent manner. There was positive correlation between Eos apoptosis and GM-CSFRa mRNA expression. Conclusion AM may promote Eos apoptosis, increase expression of GM-CSFRα in close-dependent manner in vitro. GM- CSFRα might be one of mechanisms that AM promoting apoptosis of Eos.