新型膦酸酯类化合物DHBMGP2的免疫抑制作用  

Immunosuppressive effect of a novel phosphonate compound DHBMGP2

作　　者：陈少辉[1] 肖智勇[1] 杨日芳[1] 周文霞[1] 张永祥[1] 

机构地区：[1]军事医学科学院毒物药物研究所中药和神经免疫药理研究室,北京100850

出　　处：《中国药理学与毒理学杂志》2011年第2期176-181,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家"重大新药创制"科技重大专项(2009ZX09103-018);国家"重大新药创制"科技重大专项(2009ZX09301-002)~~

摘　　要：目的从细胞和整体动物水平研究新型膦酸酯类化合物DHBMGP2免疫抑制活性。方法分离正常BALB/c小鼠脾淋巴细胞,与DHBMGP2共培养72 h,用[3H]TdR掺入法测定淋巴细胞增殖反应;染料排斥法检测24 h细胞存活率,观察淋巴细胞毒性。雄性BALB/c小鼠用二硝基氯苯(DNCB)皮肤致敏2次(间隔1周),制备小鼠迟发型超敏反应模型,初次致敏当天开始ig给药,每天1次,连续10 d,第11天处死动物测定耳肿胀度。雌性BALB/c小鼠足跖sc注射鸡卵清蛋白(OVA)免疫2次(间隔2周),制备致敏模型,首次免疫当天开始ig给药,每天1次,连续4周,每周眼球后静脉丛采血,ELISA法检测血清OVA特异性IgG,IgG1和IgG2a抗体水平,[3H]TdR掺入法检测OVA抗原特异性T细胞反应。结果 DHBMGP2 1～100μmol.L-1体外应用可明显抑制小鼠脾淋巴细胞增殖反应,对脾淋巴细胞24 h存活率无明显影响。DHBMGP2 20,40和80 mg.kg-1可以减轻DTH模型小鼠耳肿胀度,耳肿胀度由模型组的(8.1±3.4)mg分别降低为(5.2±2.1)(,5.1±1.0)和(5.4±1.3)mg,抑制其迟发型超敏反应。ig给予DHBMGP2 40和80 mg.kg-1可以明显抑制OVA致敏小鼠抗原特异性T细胞反应,[3H]TdR掺入值由模型组的(975±46)cpm分别降低为(769±30)和(601±45)cpm,但对血清OVA特异性抗体水平无明显影响。结论 DHBMGP2体内外应用对细胞免疫反应具有抑制作用,对体液免疫反应无明显影响。OBJECTIVE To investigate the immunosuppressive activity of the novel phosphonate compound DHBMGP2 in vitro and in vivo.METHODS In vitro,splenocytes from BALB/c mice was co-cultured with DHBMGP2 for 72 h,splenocyte proliferation was mearsured by thymidine uptake assay,and the 24 h survival rate was determined using trypan blue rejection assay.Dinitrochlorobenzene（DNCB）-induced delayed type hypersensitivity（DTH） models and ovalbumin（OVA）-immunized mouse model in BALB/c mice were used to assess immunosuppressive property of DHBMGP2 in vivo.Male BALB/c mice were sensitized with 5% DNCB on the first day and then challenged by DNCB on the eighth day to induce the DTH model.Vehicle or DHBMGP2 20,40 or 80 mg·kg^-1 was administered ig on the first day,then once a day,for a total of 10 d.Mice were sacrificed to measure the ear swelling on the eleventh day.Female BALB/c mice were immunized subcutaneously with OVA on the first and fourteenth day to establish an OVA-immunized mice model.From the first day of immunization on mice were ig administered with vehicle or DHBMGP2 20,40,and 80 mg·kg^-1,once a day,for 4 weeks.Blood was collected from the veniplex behind the eyeball using glass capillary once a week,and antigen-specific antibodies in serum were measured with ELISA.The mice were sacrificed at the end of 4 weeks of drug administration.Splenocyte suspension was prepared sterilized and OVA-specific T-cell responses were measured by thymidine incorporation assay.RESULTS DHBMGP2 significantly suppressed concanavalin A（Con A）-and lipopolysaccharide（LPS）-stimulated splenocyte proliferation in vitro,but had little effect on 24 h survival of the splenocytes.In the DTH model,DHBMGP2 20,40 and 80 mg·kg^-1 significantly decreased ear swelling from 8.1±3.4 to 5.2±2.1,5.1±1.0 and（5.4±1.3）mg.After treatment with DHBMGP2 40 and 80 mg·kg^-1,OVA-specific T-cell responses in OVA-immunized mice were significantly suppressed,and the thymidine uptake was decreased from 975±46 to 769±30 and（601±45）cpm.Bu

关 键 词：膦酸酯 DHBMGP2 免疫抑制剂 超敏反应 迟发型 抗原 敏感性与特异性 

分 类 号：R967[医药卫生—药理学]