冬凌草甲素衍生物对小鼠外周淋巴细胞免疫调节功能的影响  

Effect of oridonin derivative on immunoregulation of mouse lymphocytes in vitro

作　　者：王海郦[1] 王舒蓓[1] 徐莹[1] 陈柯[1] 王颖[2] 孙蕴伟[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院消化内科,上海200025 [2]上海交通大学医学院上海市免疫学研究所,上海200025

出　　处：《内科理论与实践》2014年第2期138-142,共5页Journal of Internal Medicine Concepts & Practice

基　　金：上海卫生系统学科人才建设项目(项目编号:XBR2013125)

摘　　要：目的:研究冬凌草甲素(oridonin)衍生物HAO472对小鼠淋巴细胞增殖及分泌细胞因子的影响。方法:无菌分离小鼠肠系膜淋巴结细胞,体外以抗小鼠CD3/CD28磁珠刺激淋巴细胞增殖及诱导分化,并加入不同浓度HAO472与淋巴细胞共培养,采用细胞计数试剂盒8(CCK8)检测HAO472对淋巴细胞增殖的影响;以羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)染色结合流式细胞术(FCM)检测48 h 0.5μg/mL和1.0μg/mL HAO472对CD4+T细胞增殖的作用,FCM检测48 h 0.5μg/mL和1.0μg/mL HAO472作用后CD4+T细胞分泌干扰素γ(IFN-γ)、白介素(IL)-2、IL-17的变化。结果 :0.5、1.0、1.5、2.0μg/mL的HAO472能明显抑制抗CD3/CD28磁珠诱导的淋巴细胞增殖(P<0.05),并呈剂量和时间依赖关系;淋巴细胞存活率说明0.5μg/mL和1.0μg/mL的HAO472对小鼠淋巴细胞无较大毒性作用。0.5μg/mL和1.0μg/mL的HAO472可降低CD4+T细胞的增殖指数(P<0.05)。抗CD3/CD28磁珠刺激的CD4+T细胞中,IL-17+CD4+T细胞和IFN-γ+CD4+T细胞比例为2.177%±0.229%、6.190%±0.806%。0.5μg/mL和1.0μg/mL HAO472作用后,IL-17+CD4+T细胞比例分别为1.156%±0.419%、0.806%±0.317%,IFN-γ+CD4+T细胞比例为3.964%±0.900%、2.684%±0.839%;HAO472明显抑制了CD4+T细胞分泌IL-17、IFN-γ的水平(P<0.05)。结论:水溶性冬凌草甲素衍生物HAO472可显著抑制抗CD3/CD28磁珠刺激下的小鼠淋巴细胞体外增殖及细胞因子分泌。Objective To investigate the effect of oridonin derivatitve HAO472 on proliferation and immunologic function of murine lymphocytes. Methods Lymphocytes were prepared from mesenteric lymph node （MLN） of mouse, and were stimulated with anti-CD3/CD28 coated beads for inducing proliferation and differentiation, and then were co-cultured with different concentration of HAO472. The effect of HAO472 on proliferation of lymphoeytes was determined by cell counting kit-8 （CCK8）. Proliferation of CD4＋T lymphocytes was detected by carboxyfluorescein diacetate suecinimidyl ester （CFSE） staining combined with flow cytometry. The expression of inteIferon-5,（IFN-γ）, intedeukin-17 （IL-17）, interleukin- 2 （IL-2） were assessed by flow cytometry. Results HAO472 （0.5, 1.0, 1.5, 2.0 μg/mL） could inhibit the proliferation of T lymphocytes in a dose- and time-dependent manner （P〈0.05）. The survival rate of lymphocytes showed that HAO472 0.5,1.0 p.g/mL had little toxic effect on murine lymphocytes in vitro. HAO472 0.5 , 1.0 μg/mL could inhibit the proliferation index of CD4＋T lymphoeytes（P〈0.05） . Stimulated with anti-CD3/CD28 coated beads, the percentage of IL-17＋ CD4＋T cells was 2.177%±0.229%, and that of IFN-γ＋CD4CF cells was 6.190%＋0.806%. After treated with HAO472 0.5 and 1.0 μg/mL, the percentages of IL-17＋CD4Vr cells were 1.156%±0.419% and 0.806%±0.317%, the percentages of IFN-γ＋CD4＋T cells were 3.964%_＋0.900% and 2.684%~0.839%, respectively. HAO472 obviously reduced the expression of intracellular cytokines such as IFN-γ IL-17 （P〈0.05）. Conclusions Water-soluble oridonin derivatitve HAO472 caninhibit proliferation of T lymphocytes and expression of intracellular cytokines.

关 键 词：冬凌草甲素 淋巴细胞 干扰素Γ 白介素17 

分 类 号：R285.5[医药卫生—中药学]