蛋白激酶C在乙醇诱导人γδT细胞凋亡中的作用

梅传忠; 耿建; 李柏青

蛋白激酶C在乙醇诱导人γδT细胞凋亡中的作用

Effect of protein kinase C on the apoptosis of human γδT cells induced by ethanol

摘要

摘要: 目的: 观察蛋白激酶C(protein kinase C,PKC)在乙醇诱导人γδT细胞凋亡中的作用。方法: 用结核杆菌多肽抗原(MtbAg)刺激正常人外周血单个核细胞,获得Mtb-Ag激活的T细胞(γδT细胞为主);采用Annexin-V-FITC/PI染色流式细胞术观察不同浓度乙醇对人γδT细胞凋亡诱导情况,并观察Rottlerin (PKC抑制剂)预处理对乙醇诱导γδT细胞凋亡的影响。结果: 4.0%的乙醇作用结核杆菌抗原活化的γδT细胞4 h、12 h、24 h均可显著诱导γδT细胞凋亡(P<0.01);5.0 μmol/L和40.0μmol/L Rottlerin可显著抑制4.0%乙醇诱导γδT细胞凋亡(P<0.01),凋亡抑制率分别为75.17%和58.95%。结论: PKC途径参与乙醇诱导人γδT细胞凋亡信号传导。

Abstract: Objective: To investigate the effect of protein kinase C (PKC) on the apoptosis of human γδT cells induced by ethanol.Methods: The peripheral blood mononuclear cells of healthy donors were stimulated with the peptide antigen derived from Mycobacterium tuberculosis antigen (Mtb-Ag),to generate Mtb-Ag activated T cells (γδT cells were predominant).The apoptosis of human γδT cells induced by different concentrations of ethanol was measured by flowcytometry using Annexin-V-FITC/PI staining,and the effect of pretreatment with the PKC inhibitor Rottlerin on the apoptosis of human γδT cells induced by ethanol was also investigated.Results: The apoptosis of activated human γδT cells stimulated with Mtb-Ag were significantly induced by treatment of γδT ceils with 4.0% ethanol for 4,12 and 24 hours (P<0.01).The apoptosis of γδT cells induced by 4.0% ethanol were inhibited remarkably by 5.0 μmol/L and 40.0 μmol/L Rottlerin,the inhibitory rates of apoptosis were 75.17% and 58.95%(P<0.01),respectively.Conclusions: The results suggested the PKC signaling pathways participated in the apoptosis of γδT cells induced by ethanol.

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