半边旗有效成分5F体内外的抗炎作用

In vitro and In vivo Anti-inflammatory Effects of 5F, an Extract From Pteris semipinnata

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(iNOS)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^-1 5F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iNOS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iNOS和 COX-2表达,减少炎症因子TNF-α,IL-1β,IL-6,NO和PGE₂含量有关.

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 RAW264.7 cell 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 (iNOS) 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 iNOS 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 iNOS and COX-2, decreasing the inflammatory factors such as TNF-α, IL-1β, IL-6, NO and PGE₂.