【摘要】 目的:通过建立体内外炎症模型,观察半边旗有效成分5F的抗炎作用。方法:建立脂多糖(LPS)诱导的RAW264.7细胞炎症模型,并用不同浓度5F处理,CCK-8法检测5F的细胞毒性,采用Griess法检测上清液中一氧化氮(NO)含量,ELISA法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和前列腺素E2(PGE2)含量,Western blot法检测诱导型一氧化氮合酶(i NOS)和环氧酶-2(COX-2)表达水平。雄性ICR小鼠60只,建立巴豆油和花生四烯酸诱导的耳肿胀模型,造模后随机分为模型组、阳性组和5F组,每组10只,5F组给予100 mg·kg-15F溶液,阳性组给予地塞米松0.8 mg·kg-1或吲哚美辛1 mg·kg-1,模型组给予同体积生理盐水。各组均连续给药7 d。观察5F对小鼠耳肿胀的抑制作用。结果:5F在0~40 mg·L-1的剂量内对RAW264.7细胞无明显毒性作用,10,20,40 mg·L-1的5F可依赖性抑制LPS诱导的NO释放(P<0.01),20,40 mg·L-1的5F可以降低PGE2含量(P<0.01),40 mg·L-1的5F显著性抑制i NOS和COX-2蛋白表达(P<0.05),10,20,40 mg·L-1的5F显著降低TNF-α和IL-1β含量(P<0.01),20,40 mg·L-1的5F显著降低IL-6含量(P<0.01),100 mg·kg-1的5F对巴豆油和花生四烯酸诱导的耳肿胀都有显著抑制作用(P<0.05)。结论:5F可以抑制LPS诱导的RAW264.7细胞炎症反应,抑制巴豆油和花生四烯酸诱导的耳肿胀,其抗炎作用与抑制i NOS和COX-2表达,减少炎症因子TNF-α,IL-1β,IL-6,NO和PGE2含量有关。更多还原

【Abstract】 Objective: To investigate the pharmacological activity of ent-11α-hydroxy-15-oxo-kaur-1b-en-19-oic-acid( 5F),a compound isolated from Pteris semipinnata,in vitro and in vivo inflammatory conditions.Method: The in vitro inflammation model was established using lipopolysaccharides( LPS)-induced RAW264. 7cell and different concentration of 5F was added in the culture medium. The cytotoxicity of 5F was measured by CCK-8 method. The contents of nitric oxide( NO) was examined by Griess method,tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β),interleukin-6( IL-6) and prostaglandin E2( PGE2) were determined by ELISA method. The expression of inducible nitric oxide synthase( i NOS) and cyclo-oxygen-ase-2( COX-2) were assayed by Western blot. The in vivo anti-inflammatory effect of 5F was evaluated with croton oil or arachidonic acid-induced mouse ear edema models. Result: 5F showed no cytotoxicity on RAW264. 7 cells at the concentrations from 0 to 40 mg·L- 1.5F( 10,20,40 mg·L- 1) could markedly decreased secretions of NO,TNF-α and IL-1β( P <0. 01) in LPS-induced RAW264. 7 cells. 5F( 20,40 mg·L- 1) exhibited the inhibitory effects on PGE2 and IL-6 releases( P < 0. 01). 5F( 40 mg·L- 1) could inhibit i NOS and COX-2 expression( P <0. 05). Similarly,oral administration of 5F( 100 mg·kg- 1) for 1 week diminished mouse ear edema induced by croton oil or arachidonic acid( P <0. 05). Conclusion:5F has anti-inflammtory effects,which might be mediated by inhibiting the expression of i NOS and COX-2,decreasing the inflammatory factors such as TNF-α,IL-1β,IL-6,NO and PGE2.更多还原