目的 探讨姜黄素提高γδT细胞对神经胶质瘤细胞U251杀伤作用的机理.方法 用10名健康人外周血单核细胞(PBMC)在体外经多种细胞因子培养γδT细胞;收集培养至第7天的γδT细胞,与不同浓度(0,1.25,2.5,5,10,20 μmol/L)的姜黄素共培养,同时设无水乙醇组.用FACS法测γδT细胞表型、穿孔素、颗粒酶B、细胞凋亡率和凋亡蛋白capease-3;用LDH法测定γδT细胞对U251细胞杀伤活性.结果 γδT细胞经2.5～10μmol/L姜黄素作用后能增加γδT细胞增殖,增加穿孔素、颗粒酶B的表达,杀伤U251活性明显增强(P＜0.05).当姜黄素浓度达20μmol/L时,各组检测数据均低于对照组.低浓度的姜黄素能减低凋亡率和凋亡蛋白capease-3表达,浓度为5 μmol/L时最为明显(P＜0.05).结论 一定浓度的姜黄素能增强γδT细胞对U251的杀伤活性.γδT细胞杀伤活性增强与细胞内穿孔素和颗粒酶含量相关.一定浓度姜黄素能减少其凋亡率和capease-3含量,促进U251细胞生长.%Purpose To explore the mechanism of the cytotoxicity of γδT cells induced by curcumin to U251. Methods With 10 healthy human peripheral blood mononuclear cells(PBMC) in vitro by a variety of cytokines γδT cells in culture. Collection of culture to 7 days γδT cells with different concentrations of curcumin (0,1.25,2.5,5,10,20 μmol/L respectively) co-culture. The same time, ethanol-based groups of five re-established control,at 37 ℃ ,5％ CO2 continued to train with the FACS after 72 h measuring γδT cell phenotype, perforin, granzyme B, apoptosis rate and the apoptosis protein capease-3, with the LDH determination of γδT cells to U251 cells in vitro. Results γδT cells treated with curcumin increased after 2.5-10 μmol/L when γδT cell proliferation,as compared with the control group were significantly different(P ＜ 0.05 ). γδT cells treated with curcumin after 1.25-10 μmol/L expressed by perforin,granzyme B than the control group increased(P ＜ 0.05). The 2.5-10 μmol/L the γδT curcumin induced anti-U251 cell activity was significantly enhanced in 10 μmol/L when the cytotoxic activity of up to (62.32 ± 1.28) ,was significantly higher(37.45 ± 1.12). When the curcumin concentration of up to 20 μmol/L,when the above test data of each group were lower than the control group. Low concentrations of curcumin can reduce the apoptosis rate and apoptosis proteins capease-3 expression. When the concentration of 5 μmol/L,it was the most obvious (8.68％ and 39.64％ respectively), significantly lower than the control group( 11.25％ and 42.64％ respectively). Conclusion A certain concentration of curcumin can enhance the γδT cell killing activity of U251. γδT cell killing activity was enhanced and the intracellular content of perforin and granzyme were related. A certain concentration of curcumin can reduce the apoptosis rate and capease-3 content,promoting U251 cell growth.
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