白藜芦醇对白血病细胞系KG-1a细胞被免疫细胞杀伤敏感性的影响

Enhanced sensitivity of leukemia cell line KG-1a to activated immune cell-mediated cytolysis after treated with resveratrol

目的 探讨白藜芦醇对白血病细胞系KG-1a细胞被活化免疫细胞杀伤敏感性的影响及其作用机制.方法 用锥虫蓝染色法检测白藜芦醇对KG-1a细胞增殖的半数抑制浓度（IC50值）.分离健康人外周血单个核细胞,用IL-2和IL-15刺激活化免疫细胞.用乳酸脱氢酶（LDH）释放法检测白藜芦醇作用前后KG-1a细胞对免疫活性细胞杀伤的敏感性.流式细胞术检测活化免疫细胞表面肿瘤坏死因子相关凋亡诱导配体（TRAIL）的表达及白藜芦醇作用前后KG-1a细胞表面TRAIL受体（DR4/5和DcR1/2）的表达.结果 白藜芦醇能抑制KG-1a细胞增殖,作用24 h IC50值约为25 μmol/L.在效靶比为10∶1和20∶1时,白藜芦醇作用后活化免疫细胞对KG-1a细胞的杀伤率分别为（55.80±10.88）％和（72.31±13.06）％,高于对照组的（24.96±9.25）％和（37.93±5.21）％,差异有统计学意义（P＜0.05）.白藜芦醇作用后的KG-1a细胞,DR5的表达为（9.05±3.57）％,高于作用前的（3.110.54）％,差异有统计学意义（P＜0.05）;DcR1的表达为（13.23±3.56）％,低于作用前的（53.75±10.51）％,差异有统计学意义（P＜0.05）.阻断TRAIL后,在效靶比为10∶1和20∶1时,白藜芦醇作用后活化免疫细胞对KG-1a细胞的杀伤率分别为（35.97±6.36）％和（49.80±10.68）％,低于对照组的（52.92±6.98）％和（70.73±9.79）％,差异有统计学意义（P＜0.05）.结论 白藜芦醇可增强KG-1a细胞被活化免疫细胞杀伤的敏感性,其途径可能是通过调节TRAIL通路实现的.

Objective To explore the enhanced sensitivity of leukemia cell line KG-1a to activated immune cell-mediated cytolysis after treated with resveratrol.Methods The value of 50％ inhibition concentration （IC50） for KG-1a by resveratrol was analyzed using trypan blue staining.Peripheral blood mononuclear cells were separated,and then activated by interleukin （IL）-2 and IL-15.The sensitivity of KG-1a treated with and without resveratrol to activated immune cell-mediated cytolysis was assayed by lactate dehydrogenase （LDH）-releasing assay.The expression of tumor necrosis factor related apoptosis inducing ligand （TRAIL） on the surface of activated immune cells and its receptors （DR4/5 and DcR1/2）on the surface of KG-la were detected by flow cytometry.ResuLts Resveratrol could inhibit the proliferation of KG-1a and IC50 at 24 h was 25 mmol/L.At a ratio of 10∶1 or 20∶1 between effect and target,the cytolytic rates of treated KG-1a by activated immune cells were （55.80± 10.88）％ and （72.31 ±13.06） ％,significantly higher than （24.96±9.25） ％ and （37.93±5.21） ％ of untreated KG-1 a （P＜0.05）.The expression of DR5 on the surface of KG-1 a treated with resveratrol was （9.05±3.57） ％,significantly higher than （3.11 ± 0.54） ％ of untreated KG-1 a （P＜0.05）.Conversely,the expression of DcR 1 on the surface of treated KG-la was （13.23 ± 3.56）％,lower than （53.75 ± 10.51）％ of KG-1a （P＜0.05）.When TRAIL pathway on the surface of activated immune cells was blocked,the cytolytic rates of treated KG-1a were （35.97±6.36）％ and （49.80± 10.68）％,significantly lower than （52.92±6.98）％ and （70.73±9.79）％ of untreated KG-1a （P＜0.05） at the same ratio of effector and target.Conclusions Resveratrol could enhance cytolytic sensitivity of KG-1a by activated immune cells through TRAIL pathway.