摘要
提要:目的筛选与类脂A(lipidA)具有较高结合活性的中药,从中提取具有拮抗内毒素(lipopolysaccharide,LPS)活性的物质。方法采用生物传感技术,以lipidA为靶点,检测42种中药的水提物与lipidA的结合活性;以白鲜皮为研究对象,通过大孔吸附树脂、溶剂萃取、聚酰胺柱层析和高效液相色谱(HPLC)法,分离提取与lipidA结合活性较高的组分;动态比浊法鲎试验检测HPLC产物DPR-1～3对LPS(0.1ng/ml)的中和作用;ELISA法检测DPR-2对LPS(100ng/ml)刺激的RAW264.7细胞释放肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)的影响。结果8种中药水提物与lipidA具有较高的结合活性(RU>100arc/s);从白鲜皮中逐步分离出与lipidA结合活性较高的组分,经HPLC法获得3种物质DPR-1～3;DPR-2对LPS具有一定的中和作用,在8μg/ml浓度以上可显著抑制LPS诱导的RAW264.7细胞释放TNF-α和IL-6(P<0.05),并具有剂量效应关系。结论从42种中药中筛选出与lipidA具有较高结合活性的中药白鲜皮,并从中分离提取出抗LPS活性物质DPR-2,其抗LPS活性可能与DPR-2对LPS的中和活性有关。
Objective To screen the Chinese herbs with high affinity for lipid A and extract the anti-lipopolysaccharide （LPS） compound from densefruit pittany root-bark （DPR）. Methods Forty-two kinds of aqueous extracts of traditional Chinese medicine were screened to determine their binding to lipid A by the biosensor technology. The chemical components with high affinity for lipid A were gotten by macroporous adsorptive resins, solvent extraction, polyamide laminar analysis and high performance liquid chromatogram （HPLC） from DPR. The neutralization of HPLC products DPR1 -3 for LPS （0.1 ng/ml） was detected by kinetic turbidimetric limulus test. The release of TNF-α and IL-6 in RAW264.7 cells after exposure to LPS （100 ng/ml） were detected by ELISA. Results Eight kinds of aqueous extracts including DPR displayed high affinity for lipid A （RU 〉 100 arc/s）. Three chemical components DPR1 -3 with high affinity for lipid A were gotten. DPR-2 neutralized LPS and inhibited the release of TNF-α and IL-6 in LPS-induced RAW264.7 cells in a dose-dependent manner. Conclusion Eight herbs possess the components with high affinity for lipid A. DPR-2, extracted from DPR, shows anti-LPS activity probably by binding lipid A and neutralizing LPS.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第17期1654-1656,共3页
Journal of Third Military Medical University
基金
国家重点基础研究发展规划项目("973"项目)(2005CB522600)
国家自然科学基金(30371767)~~
关键词
白鲜皮
内毒素
生物传感器
高效液相色谱
densefruit pittany root-bark
lipopolysaccharide
biosensor
high performance liquid chromatogram
