苦参碱诱导自然杀伤细胞对白血病K562细胞杀伤的实验研究  

Effects of matrine on inducing cytotoxicity of human natural killer cells against leukemia K562 cells

作　　者：马玲娣[1] 朱志超[1] 蒋丽佳[1] 卢绪章[1] 贾祝霞[1] 周民[1] 钱思轩[2,3] 李建勇[2,3] 

机构地区：[1]南京医科大学附属常州市第二人民医院中心实验室,213000 [2]南京医科大学附属常州市第二人民医院血液科,213000 [3]南京医科大学第一附属医院江苏省人民医院血液科

出　　处：《白血病．淋巴瘤》2014年第11期664-668,共5页Journal of Leukemia & Lymphoma

基　　金：基金项目：国家自然科学基金（81101647）;南京医科大学博士后课题

摘　　要：目的 探讨中药苦参碱对人自然杀伤（NK）细胞体外杀伤白血病细胞的作用及可能的分子机制.方法 以人慢性粒细胞白血病K562细胞为靶细胞,采用CFSE/PI双染色法流式细胞术检测不同质量浓度（02、0.5、0.8 mg/ml）苦参碱处理后,人NK细胞在不同效靶比下对K562细胞的体外杀伤活性.流式细胞术分析不同浓度苦参碱处理24 h对NK细胞主要活化性受体NKG2D和抑制性受体CD158a、CD158b表达的影响及K562细胞膜上NKG2D配体MICA/B、ULBP1、ULBP 2、ULBP 3表达的改变.结果 效靶比为5∶1时,NK细胞对0.2、0.5和0.8 mg/ml苦参碱处理后的K562细胞杀伤率分别为32.8％、38.1％和40.5％,较处理前均有不同程度增高（29.2％）;但进一步增加效靶比（10：1）后,NK细胞杀伤活性改变差异无统计学意义（P＞0.05）.苦参碱处理24 h,NK细胞抑制性受体CD158a、CD158b的表达均较处理前降低,而活化受体NKG2D的表达则增高.K562细胞表面NKG2D配体ULBP1和ULBP2分子的表达也较处理前增高（平均荧光强度分别为174.33±39.93比275.67±32.88,517.6±47.97比1368.6±49.43,P＜0.05）.结论 苦参碱可增强NK细胞对白血病K562细胞的体外杀伤活性,其机制可能与NK细胞受体及配体表达调节作用有关.Objective To study the effect of matrine on inducing cytotoxicity of human natural killer （NK） cell to leukemic cells and the probable molecular mechanism.Methods The cytotoxicity of NK cells was analyzed after CFSE/PI staining at different effect-to-target cell ratios by FCM using human chronic myelogenous leukemia K562 cells as target cells after treatment with different doses of matrine.The expression of NK cells main activated receptor NKG2D and the inhibitory receptor CD158a,CD158b in NK cell,as well as the expressions of four NKG2D ligands,major histocompatibility complex class Ⅰ chain-related molecules A or B （MICA,MICB）,UL16-binding proteins （ULBP） 1,2,and 3 on K562 cells were also detected before and after treatment with matrine by FCM.Results The cytotoxicity activity of NK cells against K562 cells were 32.8 %,38.1% and 40.5 % at effect-to-target ratio of 5∶1 after 0.2,0.5 and 0.8 mg/ml matrine treatment,respectively,which were increased compared with that in untreated cells （29.2 %）.At the effect-to-target ratio of 10∶1,there was no significant alternation in NK cells cytotoxicity （P 〉 0.05）.After 24 h treatment of matrine,NKG2D expression was increased in NK cells,while there was an obvious change in the levels of CD158a and CD158b.The expression of ULBP1 and ULBP2,especially ULBP2,on K562 cells dramatically were increased after treated with matrine compared with that on untreatment cells,with the mean fluorescence intensity of 174.33±39.93 vs 275.67±32.88,517.6±47.97 vs 1 368.6±49.43,respectively （P 〈 0.05）.Conclusions Matrine could enhanced the tumor-killing activity of NK cells to K562 cells,which may berelated to the modulation of NK cell receptors and their ligands expressing on the leukemia cells via matrine treatment.

关 键 词：苦参碱 自然杀伤细胞 NKG2D配体 白血病细胞 受体 自然杀伤细胞 

分 类 号：R285[医药卫生—中药学]