白鲜皮生物碱抗内毒素活性的研究.docVIP

白鲜皮生物碱抗内毒素活性的研究 杨东1，张瑛1，郑新川2，刘鑫2，鲁永玲2，郑江2 （1. 解放军第163医院，长沙410003；2. 第三军医大学西南医院综合实验研究中心，重庆400038） 从中药白鲜皮中分离抗脂多糖/内毒素的有效成分，对分离得到的成分进行定性鉴别和体、内外抗LPS活性研究。 采用大孔吸附树脂、离子交换、聚酰胺和硅胶柱层析等方法从白鲜皮水煎液中分离有效成分，并以试管法进行定性鉴别；在体外，以动态浊度法检测该成分对LPS的中和作用；ELISA 法检测其对LPS刺激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；在体内，观察该成分对热灭活大肠埃希菌攻击小鼠的保护实验。从白鲜皮中分离得到具有抗LPS作用的生物碱类成分BXP-A。体外实验结果显示，BXP-A (0.5～1.0mg/L )可抑制LPS (0.2 (g /L) 介导的鲎试剂的凝结反应（P0.05）；BXP-A(100~200 mg/L）可显著抑制LPS(100 (g /L)诱导RAW264.7细胞释放TNF-α及IL-6（P0.05或0.01）；BXP-A(40 mg/Kg)可提高注射热灭活大肠埃希菌(1.3×1011 CFU/Kg)攻击小鼠存活率约40%。BXP-A在体外能较好地中和LPS并抑制其诱导的炎症细胞因子释放，对热灭活大肠埃希菌攻击所致脓毒症模型小鼠具有保护作用。 [关键词] 白鲜皮 内毒素 细胞因子 脓毒症 中图法分类号：R285. 5 Study on anti-lipopolysaccharide activity of alkaloid components from densefruit pittany root-bark Yangdong1, Zhang Ying1, Zheng Xinchuan2, Liu Xin2, Lu Yongling2, Zheng Jiang2 (1. No.163 Hospital of PLA，Changsha 410003, China; 2. Medical Research Center, Southwestern Hospital, Laboratory, Third Military Medical University, Chongqing 400038, China) Abstract: Objective To isolate and identify the anti-lipopolysaccharide (LPS) components from densefruit pittany root-bark and investigate its potential anti-LPS activities in vitro and in vivo. Methods The components were isolated by macroporous adsorptive resins, ion exchange, polyamide and silica gel column chromatography from the aqueous extract of densefruit pittany root-bark, and identified by tube test. The neutralization of LPS was assayed by Limulus Amebocyte Lysate (LAL) test. The inhibition of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) release induced by LPS in RAW264. 7 cells was assessed by enzyme-linked immunosorbent assay (ELISA). The protective effect was observed in mice challenged by lethal dose of heat-killed Escherichia coli. Results The alkaloids components with anti-LPS activity, BXP-A, were isolated and identified from densefruit pittany root-bark. In vitro, BXP-A (0.5～1.0mg/L ) inhibited the coagulation of LAL induced by LPS(0.2 (g /L)（P0.05）,