摘要
目的对传统中药牡丹皮(Paeonia suffruticosa Andr,PSA)抗内毒素(LPS)活性组分进行定向分离及体内外活性评价。方法利用生物传感器、硅胶柱层析、聚酰胺层析、离子交换-HPLC等技术对牡丹皮进行活性组分的定向分离。通过活性组分对LPS介导RAW264.7分泌细胞因子的抑制实验、对LPS和热灭活大肠杆菌攻击小鼠的保护实验,评价所得活性组分对内毒素的拮抗作用。结果从牡丹皮水提取物中分离到一个与Lipid A具有较高结合活性的组分(PSA-3),PSA-3在体外能抑制LPS介导的RAW264.7细胞释放TNF-α(P45μg<0.05,P90、180μg<0.01),对致死剂量LPS攻击小鼠的保护率为50%(P<0.01),对致死剂量热灭活大肠杆菌攻击小鼠的保护率为50%(P<0.05),而低结合活性PSA-1组分则不具有抑制作用,对LPS和热灭活大肠杆菌(HK-E.coli)攻击小鼠也无保护效果。结论从牡丹皮中分离到一个与Lipid A具有较高结合活性的组分,该组分在体内外对LPS均具有一定的拮抗作用。
Objective To isolate the antiendotoxin active components （LPS） from Paeonia suffrutico- sa Andr （PSA） using biosensor and purification technology and assess their bioactivities in vitro and in vivo. Methods LPS were isolated from PSA using biosensor, silica gel column chromatography, polyamide chroma- tography, and IEC-HPLC. Inhibitory effect of LPS on antiendotoxin-mediated secretion of cytokines from RAW264.7 and protective effect of LPS on mice against antiendotoxin and heat killed E. coli （ HK-E. coli） were studied. Results PSA-3, an antiendotoxin active component with a high binding activity to lipid A, isolated from PSA, could inhibit TNF-α release from LPS-mediated RAW264. ？ （ P45 μg 〈 0.05, P90,180 μg 〈 0.01 ）. The protective efficiency of PSA-3 on mice was 50% against lethal LPS dose （P 〈 0.01 ）and lethal HK-E. coli dose, respectively（P 〈0.05）. PSA-I with a lower binding activity to lipid A had no inhibitory and protective effect on mice against LPS and HK-E. coli. Conclusion PSA-3, a component isolated from PSA, activity to lipid A and an antagonistic effect on LPS both in vivo and in vitro. has a high binding
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第14期1434-1437,共4页
Journal of Third Military Medical University
关键词
生物传感器
内毒素
牡丹皮
脓毒症
affinity biosensor
lipopolysaccharide
Paeonia suffruticosa Andr： sepsis
