類黃酮成分(Flavonoids)存在於許多天然植物中,目前經研究已知有抗氧化、抗發炎與抗癌等功能。而本實驗即以RAW 264.7 巨噬細胞與Thioglycollate誘導巨噬細胞之in vitro模式與Balb/c小鼠in vivo模式探討天然物Wogonin、Rutin、Quercetin對於脂多醣(Lipopolysaccharide;LPS)誘導一氧化氮(Nitric oxide;NO)、環氧化酵素第二型 (Cyclooxygenase-2;COX-2)與前列腺素E2(Prostaglandin;PGE2)引起發炎反應之抑制作用。在in vitro模式中,結果發現Wogonin與Quercetin具有明顯抑制NO的功效。Rutin則較不明顯。在in vivo模式中,我們先以尾靜脈注射給藥方式投與Wogonin、Rutin與Quercetin後再注射LPS (10 mg / kg),並且在不同時間點施以眼窩採血,血液中NO含量將以Griess reaction檢測。由實驗結果得知,經LPS誘導後,血液中NO濃度呈現time dependent方式的增加,Wogonin、Rutin、Quercetin在本模式中具有明顯NO抑制作用。但三種天然物均無較明顯之COX-2、PGE2抑制反應,僅Wogonin在RAW 264.7 巨噬細胞中產生微弱的COX-2抑制作用。綜合以上結果,我們得知Wogonin、Rutin、Quercetin具有抑制LPS 誘導NO而產生抗發炎功能,並同時建立了一個可供天然物進行發炎反應研究之實驗模式。
Flavonids are major components of natural products and several biological activities have been explored extensively including antioxidant , anti-inflammation and anti-cancer . Our present study indicated that wogonin , rutin and quercetin showed the significant inhibition on Lipopolysaccharide(LPS)-induced inducible nitric oxide synthase(iNOS) and Cyclooxygenase-2 (COX-2), Prostaglandin(PGE2)production in RAW 264.7 macrophages. As the same part of experiment, wogonin and quercetin inhibited LPS-induced iNOS gene expression and NO production in primary macrophages. Upon intravenous injection of LPS 10 mg/kg in tail vein of Balb/c mice, blood were collected from retro-orbital sinus at various time-period after injection .The amount of NO in serum was measured by griess reaction. The results appeared that the amount of NO in serum was increased in a time dependent manner in LPS-treated mice, but not in control group. Pre-treatment of Balb/c mice with Nw-nitro-L-arginine methyl ester(L-NAME)followed by LPS treatment inhibited LPS induced NO induction. Pre-treatment of Balb/c mice with wogonin, rutin and quercetin, at the concentration, was able to suppress LPS induced NO production. However, no effect was detected on LPS-induced PGE2 production. Supression of LPS induced iNOS gene expression by wogonin and quercetin was demonstrated in RAW 264.7 macrophages, primary peritoneal macrophages and liver specimen of Balb/c mice. This study firstly provided in vivo evidences to identify the inhibitory activities of wogonin, rutin and quercetin on LPS-induced NO and PGE2 production