冬虫夏草菌菌丝体提取物对 LPS诱导的炎症反应的抑制作用-微生物学专业论文.docxVIP

μg/mL,作用时间为 12h 时（相对于 6h，24h），为样品对 COX-2 的下调作用 最强；在一定浓度范围内（50,100μg/mL）,样品均可下调 COX-2 的产生并呈 现量效关系。以冬虫夏草水提物为样品研究 p38 蛋白发现浓度为 100μg/mL， 样品对 p38 的量没有影响，处理时间为 15min 时， p-p38 的下调作用最强（相 对于 30min）；在一定浓度范围内（25，50,100μg/mL）,样品对于 p38 没有影 响，均可下调 p-p38 的产生并呈现量效关系。 综上所述，冬虫夏草菌丝体提取物具有明显的体外抗炎作用。 关键词：冬虫夏草菌丝体提取物;LPS 炎症模型;小鼠腹腔巨噬细胞;炎症因子 Abstract The study is based on the inflammation model in which lipopolysaccharide (LPS) is used to stimulate mouse peritoneal macrophage RAW264.7. It studies the in vitro inflammation effects of extracts from Ophiocordyceps sinensis which is proved to be the real anamorph of precious traditional Chinese herbal medicine——Cordyceps. Meanwhile, the anti-inflammation mechanism of aqueous extract is pilot studied. This research provide theoretical and practical basement to further in vivo study, clinical study , clinical medicament as well as the exploitation of Ophiocordyceps Sinensis market. In this study the cytotoxic effects of six extracts(ethanol extract, aqueous extract and fractionated extracts of ethanol extract—— petroleum ether extract, chloroform extract, acetic ether extract ) and the residual on mouse peritoneal macrophage are examined by MTT in a proper concentration range. Then the impact on nitric oxide releasing is determined by Griess method. Western Blot is used to examine both time-effect and dose-effect of aqueous extract inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), p38 in MAPK pathway and p56 in NF-κB pathway in LPS-stimulated macrophage. The results show that OS extracts has no cytotoxicity on LPS-stimulated macrophage. Ethanol extract and aqueous extract could inhibit iNOS production in dose-dependent manner in concentrtion of 10,100,250,500μg/mL. The inhibition of chloroform extract is the strongest of all the fractionated extracts in 100μg/mL and it is the only extract that could inhibit NO production in 10μg/mL. In time-dependent reaserch, aqueous extract could inhibit iNOS, COX-2 and p3S production most in 6h,1