糖酵解抑制剂3-溴丙酮酸对异基因小鼠混合培养脾细胞增殖的影响及其机制初步研究  

Effect of glycolytic inhibitor 3-BrPA on the proliferation and apoptosis of mouse splenic lymphocytes in mixed lymphocytes culture

作　　者：周睿卿[1] 王静[2] 邱大发[1] 牛晓敏[1] 郭子文[1] 何慧清[1] 许晓军[1] 

机构地区：[1]中山大学附属中山医院血液内科,中山市528403 [2]南方医科大学

出　　处：《中华血液学杂志》2014年第7期637-640,共4页Chinese Journal of Hematology

基　　金：广东省科技计划项目[粤科社字（2011）106号];中山市科技计划重大专项（20113A002）

摘　　要：目的 研究糖酵解抑制剂3-溴丙酮酸（3-BrPA）对单向混合培养的异基因小鼠脾细胞增殖的影响及其机制.方法 以BALB/c（H-2d）小鼠脾细胞为刺激细胞,以C57BL/6（H-2b）小鼠脾细胞为反应细胞,建立单向混合淋巴细胞培养（MLC）体系,给予不同浓度3-BrPA（0～200 μmol/L）处理后,采用CCK-8法检测淋巴细胞增殖活性,流式细胞术检测MLC体系中反应细胞CD3、CD4、CD8等表面标志表达,ELISA法检测MLC体系上清中IL-4、IFN-γ等细胞因子水平.结果 CCK-8检测结果显示,高浓度（20 μmol/L以上）3-BrPA可抑制MLC体系中的T细胞增殖,并呈剂量依赖性,培养24、36、48 h的半数抑制浓度（IC50值）分别为48.6、41.2和41.9 μmol/L.50 μmol/L 3-BrPA处理24 h后MLC体系上清中IFN-γ的浓度为（164.25±20.14）ng/L,明显低于对照组[（277.61±18.46）ng/L] （P＜0.05）;IL-4的浓度为（31.06±6.06）ng/L,高于对照组[（28.64±3.97） ng/L],IFN-γ/IL-4比值明显降低.结论 浓度为20和50 μmol/L的3-BrPA对体外异基因MLC体系小鼠T细胞增殖有抑制作用,使Thl/Th2细胞平衡向Th2细胞偏移.Objective To study the effect of glycolytic inhibitor 3-Bromopyruvate （3-BrPA） on the proliferation and apoptosis of mouse spleen lymphocytes and explore its mechanism.Methods An one-way mixed lymphocyte culture （MLC） system was established,including BALB/c mouse spleen cells （H-2d） as stimulator and C57BL/6 mouse spleen cells （H-2b） as responder.With treatment of 3-BrPA at different concentrations （0-200 μmol/L）,lymphocyte proliferation capacity was detected by the CCK-8 method,the expression of CD3,CD4,and CD8 by flow cytometry,and the concentrations of cytokine interleukin （IL）-4 and interferon （IFN）-γ in the supernatant by ELISA.Results At a middle or high dose （over 20 μmol/L）,3-BrPA displayed a dose-dependent inhibitory effect on lymphocyte proliferation in the MLC system.The 50％ inhibitory concentration （IC50） were 48.6,41.2,and 41.9 μmol/L after 24,36,and 48 h culture,respectively.With treatment of 50 μmol/L 3-BrPA,the IFN-γ level [（164.25±20.14） ng/L] was significantly lower,compared with control [（277.61 ± 18.46） ng/L].The IL-4 level [（31.06 ± 6.06）ng/L] was significantly higher,compared with control [（28.64 ± 3.97） ng/L].Consequently,the IFN-γ/IL-4 ratio decreased significantly.Conclusions These results indicate that 3-BrPA had a significant inhibitory effect on the proliferation of mouse spleen lymphocytes cultured in MLC system,accompanied with the Th2-biased secretion of cytokines.

关 键 词：糖酵解抑制剂 淋巴细胞培养试验 混合 白细胞介素4 干扰素-Γ 

分 类 号：R965[医药卫生—药理学]