苗药黑骨藤多糖部位HGT-5A的细胞免疫抑制作用及其可能的活性成分  

Cellular immunosuppression effect of polysaccharides HGT-5A from Periploca forrestii Schlecht and its possible active components

作　　者：张令令[1] 肖智勇[1] 马渊[1] 齐春会[1] 周文霞[1] 张永祥[1] 乔善义[2] 孙磊[2] 

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 [2]军事医学科学院毒物药物研究所植物化学研究室,北京100850

出　　处：《中国药理学与毒理学杂志》2011年第6期538-542,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家"重大新药创制"科技重大专项(2009ZX09103-361);国家"重大新药创制"科技重大专项(2009ZX09301-002)~~

摘　　要：目的观察苗药黑骨藤多糖部位HGT-5A的细胞免疫抑制作用及探讨其可能的活性成分。方法采用二硝基氯苯建立小鼠迟发型超敏反应模型,初次致敏当天开始ig给予HGT-5A,每天1次,连续10 d,第11天处死小鼠测定耳肿胀度,观察HGT-5A体内对细胞免疫反应的影响;用[3 H]TdR掺入法检测HGT-5A及其多糖组分对小鼠脾细胞增殖反应的影响;用MTT法检测HGT-5A对脾淋巴细胞存活的影响。结果HGT-5A 50和100 mg.kg-1可以明显抑制迟发型超敏反应模型小鼠耳肿胀,耳肿胀度由模型组的(8.9±2.2)mg分别降低至6.4±1.7和(7.1±1.5)mg;HGT-5A 50～500 mg.L-1体外应用可促进小鼠脾细胞自发增殖反应,但在HGT-5A 100 mg.L-1以上时可明显抑制刀豆蛋白A诱导的小鼠脾细胞增殖反应(P<0.05),对脂多糖诱导的小鼠脾细胞增殖反应无明显影响;HGT-5A与脾细胞共培养24,48和72 h对脾细胞存活无明显影响。从HGT-5A分离获得的中性糖部位HP1,酸性糖部位HP2,从HP1中分离得到的多糖成分HP1-3,以及从HP2中分离得到的多糖成分HP2-3和HP2-4 0.5～50 mg.L-1可促进脾细胞自发增殖反应;HP2,HP1-3,HP1-4,HP2-2和HP2-4明显抑制刀豆蛋白A诱导的T细胞增殖反应。结论 HGT-5A可抑制T细胞活化增殖,对细胞免疫反应具有抑制作用,多糖成分HP1-3,HP1-4,HP2-2和HP2-4可能是黑骨藤发挥免疫抑制作用的活性成分。OBJECTIVE To investigate the cellular immunosuppression effect of polysaccharide part HGT-5A from Periploca forrestii Schlecht and its possible active components.METHODSIn vivo,the dinitrochlorobenzene（DNCB）-induced delayed type hypersensitivity（DTH） model was employed.Male BALB/c mice were sensitized with 5% DNCB on the 1st day and then challenged by DNCB on the 8th day to induce DTH.HGT-5A 50,100 or 200 mg·kg-1 was ig administered from the 1st day,once a day,for 11 d.The mice were sacrificed on eleventh day after drug administration to measure the ear swelling on the 11th day.In vitro,the splenocytes from BALB/c mice were co-cultured with HGT-5A 50-500 mg·L-1 or its polysaccharide ingredients 0.5-50 mg·L-1 for 72 h,then cell proliferation was measured by TdR uptake assay.In addition,splenocyte survival was detected with MTT assay.RESULTS Compared with model group,HGT-5A 50 and 100 mg·kg-1 significantly inhibited DTH by decreasing ear swelling from 8.9±2.2 to 6.4±1.7 and（7.1±1.5）mg.HGT-5A enhanced the primary proliferation of splenocytes,but significantly suppressed concanavalin A（Con A） stimulated lymphocyte proliferation in vitro（P〈0.05）.Splenocytes co-cultured with HGT-5A 50-500 mg·L-1 for 24,48 and 72 h manifested little effect on splenocyte survival.The neutral and acid polysaccharide parts HP1 and HP2 from HGT-5A,the polysaccharrides HP1-3 from HP1,and HP2-3 and HP2-4 from HP2 enhanced the primary splenocyte proliferaction.HP2,HP1-3,HP1-4,HP2-2 and HP2-4 obviously inhibited the Con A induced T cell proliferation.CONCLUSION HGT-5A shows immunosuppressive activity on cellular immune response,and its ingredients HP1-3,HP1-4,HP2-2 and HP2-4 may contribute partly to the immunosuppression activity of HGT-5A.

关 键 词：黑骨藤 多糖 超敏反应 迟发型 免疫抑制 

分 类 号：R285[医药卫生—中药学] R963[医药卫生—中医学]