摘要
目的观察地骨皮提取物（CL-5）对内毒素（LPS）的体内外拮抗活性。方法应用生物传感器技术检测CL-5与LPS的活性中心类脂A（Lipid A）的结合活性,ELISA法检测不同浓度CL-5（50、100、200mg/L）对LPS（100μg/L）刺激RAW264.7细胞释放肿瘤坏死因子-α（TNF-α）的影响,进一步观察不同剂量（30、60、90mg/kg）的CL-5对致死剂量热灭活大肠杆菌（E.coil）攻击小鼠的72小时存活率。结果10～640mg/L的CL-5,特异性结合值（RU）为36.5～481.2arc/s,随CL-5浓度递增而升高;单纯LPS刺激RAW264.7细胞（对照组）释放的TNF-α为（2 479.03±61.43）ng/L,给予50、100、200mg/L CL-5预处理后TNF-α浓度依次为（2 210.29±70.95）、（1 998.27±111.24）、（1 423.28±86.35）ng/L,随CL-5剂量递增而减少,均显著低于对照组（P〈0.05）;以1.30×1011CFU/kg热灭活E.coil攻击小鼠（对照组）72小时动物死亡率为100%,而以30、60、90mg/kg CL-5处理后,小鼠72小时存活率分别为50%、60%和80%,随CL-5注射剂量的增加而升高,均显著高于对照组（P〈0.05）。结论CL-5与Lipid A有一定的结合活性,可抑制LPS诱导的RAW264.7细胞活化,并对致死剂量热灭活E.coil攻击的小鼠具有保护作用。
Objective To investigate the biological activity of CL-5 in antagonizing endotoxin or lipopolysaccharide（LPS） in vitro and in vivo.Methods The binding effect to Lipid A of CL-5 was detected by the biosensor technology.The release of TNF-α in RAW264.7 cells after exposure to LPS（100μg/L） with or without CL-5 pretreatment was measured by ELISA.Furthermore,the protective effect on mice subjected to lethal dose of heat-killed E.coli challenge with or without CL-5 treatment was observed.Results The RU value of CL-5（10～640mg/L） was 36.5～481.2 arc/s,with concentration increasing,the binding effect of CL-5 to Lipid A was strengthened.In RAW264.7 cells,the TNF-α level with 50,100,200mg/L CL-5 pretreatment induced by LPS（100μg/L） was 2210.29±70.95,1998.27±111.24,1423.28±86.35ng/L,respectively,lower than that without CL-5 pretreatment（2479.03±61.43ng/L）.The survival rate of mice subjected to lethal dose of heat-killed E.coil challenge with CL-5 treatment was 50%,60%,80%,respectively in a dose of 30,60,90mg/kg,however,all mice died without CL-5 treatment.Conclusion CL-5 has a binding ability to Lipid A of LPS,It can inhibit TNF-α release in RAW264.7 cells induced by LPS and have protection effect on sepsis model animal.
出处
《创伤外科杂志》
2008年第3期260-263,共4页
Journal of Traumatic Surgery
基金
国家重点基础研究发展计划资助项目(2005CB522600)
重庆市自然科学基金项目资助(2005BB5290)
关键词
内毒素
地骨皮
脓毒症
endotoxin
cortex lycii
sepsis
