摘要
目的:研究从黄芩中分离提取拮抗内毒素(LPS)组分的制备方法及其生物学活性。方法:采用大孔吸附树脂AB-8将黄芩水提物分离成若干组分,应用生物传感器技术检测各组分与LPS的活性中心LipidA的结合活性,ELISA法检测各组分对LPS刺激RAW264.7细胞释放肿瘤坏死因子α(TNF-α)的影响;由此筛选出能与LipidA结合并对LPS刺激的RAW264.7细胞活化具有抑制作用的组分,进一步观察所得组分对脓毒血症模型动物的保护作用。结果:采用大孔吸附树脂AB-8从黄芩中共分离到5种组分(SbG-1～5),其中SbG-4、SbG-5与LipidA具有较高结合活性,特异性结合值(RU)分别为113.36arc/s、73.75arc/s;SbG-4、SbG-5(100mg/L)可显著抑制LPS(100μg/L)诱导的RAW264.7细胞释放TNF-α(P<0.05);SbG-4能显著提高热灭活大肠杆菌(1.33×1011CFU/Kg)所致脓毒血症小鼠的72h存活率(P<0.05),并呈剂量效应关系。结论:从黄芩中分离提取的具有拮抗LPS活性的SbG-4可抑制LPS刺激的RAW264.7细胞活化,并对脓毒血症模型动物具有显著的保护作用。
Objective:To investigate the preparation methods of extracting anti-LPS active compounds from Scutellaria baicalensis Georgi and evaluate its biological activity.Method:A series of fraction was prepared by macroporous adsorptive resins AB-8 from the aqueous extract of Scutellaria baicalensis Georgi.Their binding activity to Lipid A and effect on the release of TNF-α in LPS-stimulated RAW264.7 cells were detected by biosensor method and ELISA respectively.Thus,the compound which has binding activity to Lipid A and inhibitory action on LPS-stimulated RAW264.7 cells activation was screened.Subsequently,its protective effect on sepsis model mice induced by heat-killed E.coli was further observed.Results:Five kinds of compounds isolated from Scutellaria baicalensis Georgi were named SbG-1～5.Among them,the binding activity of SbG-4 and SbG-5 to Lipid A were higher than the others,with their RU value being 113.36 arc/s,73.75 arc/s,respectively.Both SbG-4 and SbG-5(100 mg/L)could inhibit the release of TNF-α in LPS-induced RAW264.7 cells.Interestingly,the experimental data indicated that SbG-4 could significantly increase the survival rate of sepsis mice induced by heat-killed E.coli in a dose-dependent manner.Conclusion:SbG-4 extracted from Scutellaria baicalensis Georgi possess anti-LPS activity,which could suppress the activity of LPS-stimulated RAW264.7 cells in vitro and have protective effect on sepsis model mice in vivo.
出处
《微循环学杂志》
2008年第1期20-23,27,共5页
Chinese Journal of Microcirculation
基金
国家重点基础研究发展计划资助项目(2005CB22600)
