乙醇降低人外周血T淋巴细胞活化并增加其凋亡

Ethanol decreases activation of T lymphocytes in human peripheral blood and increases their apoptosis

目的观察乙醇对人外周血T淋巴细胞亚群数量、活化功能及凋亡的影响.方法从健康供血者通过密度离心分离出外周血单个核细胞(PBMC),淋巴细胞与植物血凝素(PHA)和(30、50)mL/L乙醇共同培养.培养12、18、36、48 h后,采用流式细胞术检测各组淋巴细胞活化功能以及凋亡情况.结果PBMC与乙醇共培养12 h后,在(30、50)mL/L的乙醇培养条件下,T淋巴细胞的数量在12 h内无明显变化,但与空白对照相比,PHA组CD8^+T细胞出现活化,而且50 mL/L乙醇可以抑制CD8^+T细胞的活化.PBMC与乙醇共培养18 h,与单纯PHA组相比,乙醇和PHA联合组淋巴细胞数量减少,且随着乙醇剂量的增加淋巴细胞数量下降更明显,且PHA刺激T细胞活化减弱.PBMC与乙醇共培养36 h,与单纯PHA组相比,乙醇联合PHA处理组淋巴细胞的数量减少更显著,共培养48 h,淋巴细胞几乎消失.随着PBMC与乙醇共培养时间的延长和乙醇剂量的增加,CD3^+T淋巴细胞凋亡比例升高.结论较高剂量乙醇处理的外周血T淋巴细胞活化减少,随着乙醇剂量的增加,T淋巴细胞及其亚群的细胞数量减少,可能与促进T淋巴细胞及其亚群凋亡有关.

Objective To detect the effect of ethanol on T lymphocyte subsets,immune activity and apoptosis in peripheral blood.Methods Peripheral blood mono nuclear cells(PBMCs)were isolated from healthy donors by density centrifugation.Lymphocytes were co-cultured with phytohaemagglutinin(PHA)in 30 and 50 mL/L ethanol.At 12,18,36,48 hours after PBMCs were exposed to alcohol,flow cytometry was performed to examine the alterations of T lymphocyte subsets.Results Compared with the control group,at 12 hours after the exposure,the number of CD8+CD69+T lymphocytes increased significantly in both PHA and alcohol groups but the number of CD8+CD69+T lymphocytes in the PHA group was apparently higher than that of 50 ml/L ethanol group,while the number of T cells had no obvious change.At 18 hours after exposed to ethanol,the proliferation of T lymphocytes was reduced obviously in the ethanol groups.In addition,the proliferation of T cells in the ethanol(50 ml/L)group was markedly lower than that in 30 ml/L ethanol group.At the same time,the number of CD8^+CD69^+T cells was markedly lower.At 36 hours after exposed to ethanol,in comparison with the PHA group,the number of T lymphocytes decreased markedly in the ethanol groups.At 48 hours after exposed to etha nol,T cells almost disappeared in the ethanol groups.The apoptosis rate of CD3^+T lymphocytes in creased with the extension of co-culture time of PBMCs and ethanol as well as the increase of ethanol dose.Conclusion With the in crease of ethanol dose,the activity of T lymphocytes in the peripheral blood treated with higher dose of ethanol would go down and the number of T lymphocyte subsets and the function of T cells decrease,which might be related to the ethanol that promotes the apoptosis of T lymphocyte subsets.