【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。更多还原

【Abstract】 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-β and IFN-γ in 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-γ and TNF-β release were significantly increased after oridonin treatment(P<0.05), however, the TNF-α release 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-γ and TNF-β release.更多还原