板蓝根F022部位抗内毒素分子机理研究_药学论文.docVIP

板蓝根F022部位抗内毒素分子机理研究_药学论文 板蓝根F022部位抗内毒素分子机理研究_药学论文 作者：林爱华，刘云海，刘奕明 【关键词】 板蓝根；,,F022部位；,,内毒素；,,分子机理 　　摘要：目的探讨板蓝根F022部位抗内毒素分子机理。方法内毒素定量检测法测定F022样品液对内毒素的破坏作用；研究样品液对内毒素致鼠巨噬细胞分泌炎性因子的抑制作用、对蛋白激酶MAPKp38活性的影响、对内毒素刺激鼠组织moesin mRNA分子表达及对内毒素诱导鼠体内TNFα,IL6和NO的抑制作用。结果1%F022对内毒素破坏率达到86.5%，F022能显著抑制内毒素刺激巨噬细胞分泌TNFα，IL6水平；抑制内毒素刺激蛋白激酶MAPKp38活性；降低内毒素刺激鼠肝、肾、脾组织moesin mRNA和体内TNFα，IL6和NO等炎性因子的表达。结论F022部位不仅直接破坏内毒素结构，且阻断内毒素引起的信号传导通路，从而抑制机体炎性因子的过度释放、抑制膜结构伸展刺突蛋白和丝裂原活化蛋白激酶的表达，从分子水平阐明了板蓝根中活性部位F022抗内毒素的分子机理。 　　关键词：板蓝根； F022部位； 内毒素； 分子机理 　　Studies on the Antiendotoxic Molecular Mechanism of F022 from Radix Isatidis 　　Abstract：ObjectiveTo study the anti-endotoxic molecular mechanism of F022 from Radix Isatidis. MethodsThe content of F022pretreated ET was quantitatively determined with limulus test．The inhibition of TNFα and IL6 of murine peritoneal macrophages stimulated by LPS,the molecular expression of moesin mRNA, MAPK p38, TNFα,IL6 and NO in mice tissues induced by LPS were studied on the antiendotoxic mechanism of F022. ResultsIt was found that the ET reduction rate was 86.5%, if 1%F022was added to macrophages culture before addition of 50 ng#12539;ml 1LPS．Production of TNFα, IL6 and MAPKp38 by macrophages were inhibited in vitro．F022 remarkably decreased the molecular expression of moesin mRNA in liver，kidney，spleen induced by LPS in mice and inhibited production of TNFα, IL6 and NO in mice stimulated by LPS.ConclusionF022 from Radix Isatidis can not only destroy the structure of endotoxin, but also obstruct the signal transduction pathways of LPS. F022 may be a strong antiendotoxin fraction in drugs research and development. 　　Key words：Radix Isatidis; F022 fraction; Endotoxin; Molecular mechanism 　　内毒素(endotoxin，ET)是革兰氏阴性菌细胞壁外膜中的脂多糖(1ipopolysaccharide，LPS)成分。可刺激机体防御系统过度释放炎性因子肿瘤坏死因子(TNFα)、白细胞介素6(IL6)及一氧化氮(NO)等而引起发热、脓毒性休克、弥漫性血管内凝血(DIC)、多器官功能衰竭综合征(MODS)，以至死亡。中医对内毒素性疾病的早期表现常辨证为“实热证”，内毒素是“热毒、邪毒”的物质基础［1］。有关内毒素的研究一直是世界生物医学研究热点，而内毒素拮抗药的研究则是防治内毒素性疾病的重要策略之一。先前研究已证实板蓝根有抗大肠杆菌O111B4内毒素作用［2，3］，我们从中筛选出抗