黄芩苷对BCG感染后巨噬细胞表达NKG2 D配体及NK细胞杀伤活性的影响  

Baicalin promotes the expression of NKG2D ligands on BCG-infected macrophages and enhances the cytotoxic activity of NK cells

作　　者：车芸华 王莉新[1] 王易[1] 刘丹[1] 

机构地区：[1]上海中医药大学免疫学与病原生物学教研室,201203

出　　处：《中华微生物学和免疫学杂志》2016年第9期647-653,共7页Chinese Journal of Microbiology and Immunology

基　　金：上海市卫生和计划生育委员会科研课题（20124182）;上海市自然科学基金（13ZR1441800）;教育部高等学校博士学科点专项科研基金（20133107120011）：国家自然科学基金（81401300）

摘　　要：目的：探讨卡介苗（BCG）感染对巨噬细胞表面NKG2D配体（MICA、MICB、ULBP1和ULBP2）表达的影响，以及黄芩苷对配体表达及NK细胞杀伤活性的影响。方法佛波醇酯（ PMA）诱导THP-1细胞分化为巨噬细胞，建立BCG感染巨噬细胞模型，进一步用黄芩苷干预感染模型，分别采用real-time PCR和Western blot检测MICA、MICB、ULBP1和ULBP2 mRNA水平和蛋白表达水平。人外周血单个核细胞（ PBMC）诱导分化扩增的NK细胞与感染后巨噬细胞共孵育4 h后，采用RTCA DP实时无标记细胞分析仪检测NK细胞的杀伤活性。结果 BCG感染可上调MICA、MICB、ULBP1和ULBP2 mRNA水平和MICA、ULBP1蛋白表达水平。1 mg/L黄芩苷处理72 h后可显著促进MICA和ULBP1 mRNA和蛋白表达，并增加NK细胞对BCG感染后巨噬细胞的杀伤率。结论 BCG感染可诱导人巨噬细胞NKG2D配体表达增加，但不能有效活化NK细胞；黄芩苷可进一步上调感染后巨噬细胞表达NKG2D配体，进而促进NK细胞杀伤活性。Objective To investigate the effects of BCG （ Bacillus Calmette-Guerin） infection on NKG2D （natural killer group 2, member D） ligands （MICA, MICB, ULBP1 and ULBP2） expressed on macrophages and to further analyze the effects of baicalin on these NKG2D ligands and the cytotoxic activity of NK cells. Methods PMA （ phorbol 12-myristate 13-acetate） was used to induce the differentiation of THP-1 cells into macrophages. The THP-1-derived macrophages were infected with BCG and then treated with baicalin. The expression of MICA, MICB, ULBP1 and ULBP2 at mRNA and protein levels were meas-ured by real-time PCR and Western blot assay. The BCG-infected macrophages were co-cultured with NK cells derived from human PBMC for 4 h. Real-Time Cell Analyzer （ RTCA DP） was used to evaluate the cy-totoxic activity of NK cells. Results The expression of MICA, MICB, ULBP1 and ULBP2 at mRNA level and the expression of MICA and ULBP1 at protein level were upregulated after infecting the macrophages with BCG. The expression of MICA and ULBP1 at mRNA and protein levels and the killing activity of NK cells were significantly enhanced after treating the BCG-infected macrophages with baicalin （1 mg/L） for 72 h. Conclusion BCG infection could induce the expression of NKG2D ligands on human macrophages, but could not effectively active the NK cells. Baicalin could enhance the cytotoxic activity of NK cells by further up-regulating the expression of NKG2D ligands on BCG-infected macrophages.

关 键 词：黄芩苷 结核分枝杆菌 巨噬细胞 NK细胞 配体 

分 类 号：R392[医药卫生—免疫学]