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目的:在前期研究的基础上进一步观察氨茶碱对人外周血T淋巴细胞凋亡与胀亡的诱导作用。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,分空白组及氨茶碱组,培养后观察细胞光镜及电镜形态学、FDA/PI荧光染色,并用流式细胞仪检测凋亡和胀亡细胞比例变化。结果:①人外周血T淋巴细胞经体外培养,可自然出现典型的细胞凋亡与胀亡形态学改变。②人外周血T淋巴细胞在氨茶碱诱导作用下,凋亡率有显著增高,但胀亡率相对无显著增高。结论:人外周血T淋巴细胞存在凋亡与胀亡现象,氨茶碱可诱导人外周血T淋巴细胞凋亡,但不能有效的诱导健康人外周血T淋巴细胞胀亡。
OBJECTIVE: To further observe the induction effect of aminophylline on the apoptosis and expansion of T lymphocytes in human peripheral blood on the basis of previous studies. METHODS: Peripheral blood T lymphocytes were isolated from healthy adults by density gradient centrifugation and nylon cotton column method. The cells were divided into blank group and aminophylline group. The morphological changes of cells were observed by light microscopy, electron microscopy and fluorescence staining with FDA / PI. Cytometry was used to detect the ratio of apoptotic and inflamed cells. Results: (1) T lymphocytes from human peripheral blood were cultured in vitro, and typical morphological changes of apoptosis and bulging cells were observed. ②Apoptosis rate of human peripheral blood T lymphocytes was significantly increased under the induction of aminophylline, but the relative rate of expansion and death was not significantly increased. Conclusion: Apoptosis and expansion of T lymphocytes in human peripheral blood are observed. Aminophylline can induce T lymphocyte apoptosis in human peripheral blood, but it can not effectively induce the expansion of T lymphocytes in healthy human.