氨茶碱诱导T淋巴细胞自噬现象的观察

Autophagy induced by aminophylline in peripheral blood T lymphocytes

目的:观察氨茶碱对分离培养的人外周血T淋巴细胞自噬的影响。方法:密度梯度离心法及尼龙棉柱法分离健康成年人外周血T淋巴细胞,分对照组、氨茶碱组(10-8～10-3mol/L,即0.0018～180μg/ml)及地塞米松组(DXM),培养后用流式细胞术检测自噬率和凋亡率变化。结果:①T淋巴细胞的分离纯度为81.3%～94.5%。②氨茶碱组干预T淋巴细胞培养72小时后,10-5～10-3mol/L氨茶碱组自噬率、10-8～10-3mol/L氨茶碱组凋亡率与阴性对照组比较差异均有显著性(P值均<0.05);各浓度组的自噬率和凋亡率之间均无相关性(P值均>0.05);在10-5mol/L氨茶碱干预下,按不同细胞密度接种T淋巴细胞,观察0、24、48、72小时的自噬率,可见随着细胞密度的减少和时间的延长,其自噬率有增加趋势,但无显著性差异。③DXM干预T淋巴细胞72小时后,其凋亡率与阴性对照组比较有显著差异(P=0.000),而自噬率无显著差异(P=0.481)。结论:10-5～10-3mol/L的氨茶碱可诱导人外周血T淋巴细胞自噬率增加,10-8mol/L～10-3mol/L的氨茶碱可诱导人外周血T淋巴细胞凋亡率增加,且自噬与凋亡间相互独立;DXM可诱导人外周血T淋巴细胞凋亡率增加,表明不同的药物可诱导T淋巴细胞启动不同的程序性死亡方式。

Objective：To observe autophagy induced by aminophylline in human peripheral blood T lymphocytes.Methods：Peripheral blood T lymphocytes from health adults were separated with Percoll（1.073 g/ml） and harvested by using nylon column.The cultured cells were divided into control group,aminophylline group（10^-8～10^-3mol/L,or 0.001 8～180 μg/ml）and dexamethasone（DXM） group,and the rates of autophagy and apoptosis were analyzed by flow cytometry.Results：①The purities of T lymphocyte were 81.3%～94.5%.②In control group for 72 h cultivation,the autophagic rate of T lymphocytes was significantly lower than those of 10^-5～10^-3mol/L aminophylline group（P〈0.05）;The apoptotic rate of T lymphocytes was significantly lower than those of 10^-8～10^-3mol/L aminophylline group（P〈0.05）;There was no significant statistic difference in correlation between autophagy and apoptosis of T lymphocytes after 72 h cultivation in aminophylline group（P〉0.05）;In 10^-5mol/L aminophylline group,the autophagic rate of different T lymphocyte density after cultivation（0,24,48,72 h） had a tendency of increase along with time lasting and decrease of cell density,but the difference did not show significance.③There was significant statistic difference in incidence rate of apoptotic T lymphocytes between control and DXM group in 72 h cultivation（P=0.000）;But there was no significant statistic difference in the rate of autophagic（P=0.481）.Conclusion：10^-3～10^-5mol/L aminophylline can induce autophagy in peripheral blood T lymphocytes,10^-3～10^-8mol/L aminophylline could induce both autophagy and apoptosis,and there is no significant correlation between autophagy and apoptosis;DXM can induce apoptosis in peripheral blood T lymphocytes,which indicates different programmed cell death of T lymphocytes could be induced by different medicines.