补体抑制活性物质鱼腥草总多糖中内毒素的去除Removal of lipopolysaccharides from anti-complementary crude polysaccharides of Houttuynia Herba
张娟娟;张庆琳;卢燕;陈道峰;
ZHANG Juan-juan;ZHANG Qing-lin;LU Yan;CHEN Dao-feng;School of Pharmacy,Fudan University;
摘要(Abstract):
为建立补体抑制活性鱼腥草总多糖中内毒素的去除方法,将琼脂糖疏水色谱及聚丙烯酰胺凝胶过滤色谱分别与多粘菌素亲和吸附色谱联用纯化鱼腥草多糖。以显色基质比色法检测纯化前后内毒素的含量变化,细胞溶血法检测纯化前后体外抗补体活性变化。结果发现,疏水色谱与亲和吸附色谱的联用对内毒素的清除效果优于凝胶过滤色谱与亲和吸附色谱的联用,其清除率达42.85%。内毒素清除前后,抗补体活性没有明显变化。表明疏水色谱与亲和吸附色谱联用可用于清除鱼腥草总多糖中的内毒素且不影响其抗补体活性。
An Affi-Prep Polymyxin column was combined with a Phenyl Sepharose column and a Sephacryl S-300 column,respectively,to remove the lipopolysaccharides( LPS) in the anti-complementary crude polysaccharides of Houttuynia Herba. The contents of LPS in the polysaccharides were determined by chromogenic tachypleus amebocyte lysate( TAL) method during the procedure of purifying. The anti-complementary activities of the polysaccharides were also compared before and after the removal of LPS. Less remanent LPS was detected after purified using Penyl Sepharose combined with polymyxin column,with the clearance rate of 42. 85%. All the columns had no effect on the anti-complementary activity of the polysaccharides. Penyl Sepharose combined with polymyxin column would be sound for LPS removal of the anti-complementary polysaccharides without reducing their bioactivity.
关键词(KeyWords):
鱼腥草总多糖;内毒素;抗补体;疏水色谱;亲和吸附色谱
crude polysaccharides of Herba Houttuynia;LPS;anti-complementary;Penyl Sepharose;polymyxin
基金项目(Foundation): 国家“重大新药创制”科技重大专项(2012ZX09301001-003);; 国家自然科学基金项目(81330089,30801508)
作者(Author):
张娟娟;张庆琳;卢燕;陈道峰;
ZHANG Juan-juan;ZHANG Qing-lin;LU Yan;CHEN Dao-feng;School of Pharmacy,Fudan University;
Email:
DOI: 10.19540/j.cnki.cjcmm.20170728.011
参考文献(References):
- [1]Wu Y S,Chen Y Y,Ueng P S,et al.Effects of medicinal herbs"Plantago asiatica","Houttuynia cordata"and"Mentha haplocalyx"on non-specific immune responses of cobia(Rachycentron canadum)[J].Fish Shellfish Immun,2016,58:406.
- [2]许贵军,李志军,王琦,等.鱼腥草的抗炎活性成分[J].中国药科大学学报,2016,47(3):294.
- [3]Sekita Y,Murakami K,Yumoto H,et al.Anti-bacterial and antiinflammatory effects of ethanol extract from Houttuynia cordata poultice[J].Biosci Biotechnol Biochem,2016,80(6):1205.
- [4]Werner J.Functional mechanisms of temperature regulation,adaptation and fever:complementary system theoretical and experimental evidence[J].Pharm Thera,1988,37(1):1.
- [5]Jiang Y,Lu Y,Zhang J J,et al.Anti-complementary constituents of Houttuynia cordata and their targets in complement activation cascade[J].Nat Prod Res,2014,28(6):407.
- [6]Wu M,Li H,Zhang H,et al.Development of a C3c-based ELISA method for the determination of anti-complementary potency of Bupleurum polysaccharides[J].Acta Pharmacol Sin,2015,5(4):316.
- [7]姚士,徐乃玉,陈道峰,等.蓝萼香茶菜的抗补体活性成分研究[J].中国中药杂志,2013,38(2):199.
- [8]阮姝楠,卢燕,陈道峰.广藿香的抗补体活性成分[J].中国中药杂志,2013,38(13):2129.
- [9]张娟娟,卢燕,陈道峰.鱼腥草抗补体活性多糖的制备工艺研究[J].中国中药杂志,2012,37(14):2071.
- [10]Xu Y Y,Zhang Y Y,Ou Y Y,et al.Houttuynia cordata Thunb.polysaccharides ameliorates lipopolysaccharide-induced acute lung injury in mice[J].J Ethnopharmacol,2015,173:81.
- [11]陈道峰,姜韵,卢燕,等.鱼腥草总多糖及其制备方法和在制药中的用途:中国,ZL201010022880.4[P].2010-10-02.
- [12]Wyns H,Plessers E,Backer P D,et al.In vivo porcine lipopolysaccharide inflammation models to study immunomodulation of drugs[J].Vet Immunol Immunopathol,2015,166(3/4):58.
- [13]邓仲端,瞿智玲,杨丽敏.内毒素脂多糖诱导培养的人脐静脉内皮细胞表达巨噬细胞炎性蛋白-1α[J].中华病理学杂志,2003,32(5):51.
- [14]李荣芳,李晓文,王吉,等.细菌脂多糖及其免疫应激研究进展[J].中兽医医药杂志,2015(4):23.
- [15]王雪薇,王建华.Triton X-114萃取法去除脑膜炎球菌荚膜多糖中的内毒素[J].中国生物制品学杂志,2008,21(5):438.
- [16]李友,龚慕辛,关怀,等.动态浊度法比较不同方法对内毒素的清除效率[J].中国医院药学杂志,2010,30(14):1175.
- [17]周康,刘康达,陈梅芳,等.去除内毒素的研究及临床应用(一种新型亲和层析膜内毒素清除初步应用)[J].镇江医学院学报,1998,8(4):465.
- [18]张顺财,朱无难,周康,等.多粘菌素B琼脂糖层析柱清除体液中内毒素[J].上海医科大学学报,1992,19(1):9.
- [19]唐守万,孔亮,欧俊杰,等.柱亲和介质用于内毒素去除的研究[J].分析化学研究报告,2006,34(4):455.
- [20]商振华,周冬梅,郭为,等.去除内毒素的柱亲和介质与膜亲和介质的制备与特征[J].分析化学,1997(9):1010.
- [21]曹春雨,刘婷,郭静,等.显色基质法检测6种中药注射液内毒素含量[J].中国实验方剂学杂志,2010,16(9):88.
- [22]李贺,张培因,卫红飞,等.重组卡介苗热休克蛋白70的纯化及内毒素去除的效果评价[J].吉林大学学报:医学版,2010,36(1):35.
- [23]王贻杰,刘勇,程海,等.提高内毒素去除效果的质粒DNA层析纯化工艺的优化[J].中国生物制品学杂志,2009,22(1):81.
- [24]郑丽君,姚勇,何国振.利用纯化工艺去除降纤酶细菌内毒素[J].广东药学,1999,9(2):29.
- [25]朱晓丽,范代娣,马戎.阴离子交换法去除类人胶原蛋白中的内毒素[J].化学工程,2011,39(3):67.
- [26]李存玉,郑云枫,李红阳,等.细菌内毒素定量法研究活性炭与超滤法热原去除工艺[J].医药导报,2010,29(2):142.
- [27]邵英光,李京华,魏桂林,等.内毒素的去除政策[J].广州化学,2003,28(2):38.
- 鱼腥草总多糖
- 内毒素
- 抗补体
- 疏水色谱
- 亲和吸附色谱
crude polysaccharides of Herba Houttuynia - LPS
- anti-complementary
- Penyl Sepharose
- polymyxin