冬凌草甲素对NK-92 MI杀伤THP1细胞的活性影响及机制  

Influence of Oridonin on the Icilling Acitivity of NK-92 MI Cells Targeting Cell THP1 and Its Mechanism

作　　者：刘雁峰 贾妍[1] 贺鹏程[2] 张梅[2] 何群[1] LIU Yan-Feng;JIA Yan;HE Peng-Cheng;ZHANG Mei;HE Qun(Department of Hematology,Xiangya Hospital,Central South University,Changsha 410008,Hunan Province,China;Department of Hematology,The First Affliated Hospital,Xi'an Jiaotong University,Xi'an 710061,Shaanxi Province,China)

机构地区：[1]中南大学湘雅医院血液科,湖南长沙410008 [2]西安交通大学第一附属医院血液内科,陕西西安710061

出　　处：《中国实验血液学杂志》2019年第5期1374-1379,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金青年科学基金项目(81600135)

摘　　要：目的:观察冬凌草甲素对效应细胞NK-92 MI杀伤靶细胞THP1活性的影响并探讨其作用机制。方法:LDH释放法检测冬凌草甲素作用前后NK-92 MI对THP1的杀伤效率;应用qRT-PCR和Western blot分别检测THP1细胞表面自然杀伤细胞激活性受体D(NKG2D)相关配体表达情况;ELISA法检测效靶细胞共培养上清中细胞因子表达变化。结果:冬凌草甲素处理后NK-92 MI在不同效靶比均可提高对THP1细胞的杀伤效率;冬凌草甲素可上调THP1细胞表面MICB、ULBP1和ULBP2表达,同时增加效靶细胞共培养上清中IFN-γ和TNF-β释放,而对MICA、ULBP3表达及TNF-α释放无明显影响。结论:冬凌草甲素通过增加MICB、ULBP1和ULBP2表达以及促进IFN-γ和TNF-β释放提高THP1细胞对NK-92 MI的杀伤敏感性。Objective:To investigate the influence of oridonin on the killing activity of NK-92 MI cells targeting THP1 and the related mechanism.Methods:The killing activity of NK-92 MI to THP1 before and after oridonin treatment was detected by LDH release assay;the expression of natural killer cell ligands activating receptor D(NKG2 D,including MICA,MICB,ULBP1,ULBP2 and ULBP3)was detected by real-time quantitative polymerase chain reaction(qRT-PCR)and Western blot respectively;the expression of cytokine TNF-α,TNF-βand IFN-γin the co-culture supernatant of NK-92 MI cells and THP1 cells were measured by ELISA.Results:The killing efficiency after oridonin treatment at different effector-target ratio(1:1,5:1,10:1)was all significantly up-regulated in comparison with that before oridonin treatment(P<0.05).QRT-PCR and Western blot showed that the expressions of mRNA and protein levels of MICB,ULBP1,ULBP2 increased to varying degree(P<0.05),but the expression levels of MICA and ULBP3 were not statistically significant between experimental group and control group(P>0.05).ELISA results indicated that IFN-γand TNF-βrelease were significantly increased after oridonin treatment(P<0.05),however,the TNF-αrelease was not statistically different in comparison with control group(P>0.05).Conclusion:Oridonin can significantly improve killing efficiency of NK-92 MI on THP1,that might be related with up-regulation of MICB,ULBP1 and ULBP2 expression and promotion of IFN-γand TNF-βrelease.

关 键 词：冬凌草甲素 NK-92 MI THP1 效靶杀伤 免疫治疗 

分 类 号：R733.71[医药卫生—肿瘤] R392.33[医药卫生—临床医学]