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目的 :探讨氨茶碱对大鼠气道平滑肌细胞 (ASMCs)增殖抑制和促凋亡作用及在哮喘发生、发展中的机制 .方法分离大鼠气道平滑肌 ,在含有 10 0mL·L -1胎牛血清的DMEM中培养 ,第 4～ 6代细胞用于实验 ,细胞用不同浓度氨茶碱 (0 ,5 0 ,10 0 ,2 0 0 ,30 0 ,4 0 0mg·L -1)诱导培养12～ 72h ;用光学显微镜观察其形态学凋亡改变 ;用琼脂糖电泳分析DNA片段 .结果 :①氨茶碱以时间和浓度依赖的方式降低存活的细胞数 ;②当氨茶碱浓度 >5 0mg·L -1时 ,光镜下可见细胞核皱缩、碎裂 ,随着药物浓度增加 ,凋亡小体形成 ;③当氨茶碱浓度 >5 0mg·L -1时 ,琼脂糖电泳显示了典型的梯状条带 .结论 :实验表明氨茶碱可以诱导体外大鼠ASMCs发生凋亡样改变
Aims: To investigate the inhibitory and proapoptotic effects of aminophylline on rat airway smooth muscle cells (ASMCs) and its mechanism in the pathogenesis and development of asthma.Methods Rat airway smooth muscle was isolated and cultured in a medium containing 10 0 mL·L -1 fetal Bovine serum was cultured in DMEM. Cells from passage 4 to passage 6 were used for experiments. Cells were induced to culture with different concentrations of aminophylline (0, 50, 100, 200, 300, 400 mg · L -1) 12 ~ 72h.The morphological changes of apoptotic cells were observed by optical microscope.DNA fragments were analyzed by agarose gel electrophoresis.Results: ①Aminophylline reduced the number of viable cells in a time-and-concentration-dependent manner.②When the concentration of aminophylline was> 5 When the concentration of aminophylline was above 50 mg · L -1, agarose gel electrophoresis showed that the apoptotic bodies formed when the concentration of aminophylline was above 50 mg · L -1 Typical ladder-like ribbon.Conclusion: The experiment shows that aminophylline can induce apoptosis-like changes in rat ASMCs in vitro