本研究主要目的為探討金針花苞不同生長階段及不同萃取方法(超音波水萃、攪拌水萃、超音波乙醇萃、攪拌乙醇萃)對抗發炎活性的影響。結果顯示萃取物中以超音波水萃取物(UWE) 抑制NO生成效果較佳，其中又以UWE-stage 2效果最佳，於125 ppm可達91.55 %抑制率。其次依據ELISA分析UWE-stage 2~4對發炎巨噬細胞分泌細胞激素的影響，UWE-stage 2~4於75~125 ppm對於促發炎激素TNF-α及PGE2並無抑制作用，且會顯著增加TNF-α之生成量。相反的三個生長階段的UWE均可顯著降低促發炎激素IL-6，其中仍以UWE-stage 2抑制效果最為顯著，於125 ppm可達85.2 %抑制率。此外，在抗發炎激素IL-10方面，UWE-stage 2~4隨著劑量上升而下降，透過IL-6/IL-10比值降低的結果指出UWE-stage 2最具抗發炎潛力，比值可由4.22降至2.97。最後，以Western blot分析UWE對發炎巨噬細胞中COX-2及iNOS蛋白質表現量的影響，結果顯示UWE-stage 2抑制NO生成效果是透過抑制iNOS的表現量。 The aim of this study was to investigate the effects of different growth stages and extraction methods (ultrasonic water extraction, stir water extraction, ultrasonic ethanol extraction, stir ethanol extraction) on anti-inflammatory activity of daylily buds. The result showed ultrasonic water extract (UWE) had the better inhibition on NO production. UWE-stage 2 has the best inhibition among the extracts, up to 91.55 % at 125 ppm level. Base on ELISA analysis the effects of UWE from daylily growth stage 2, 3 and 4 with different dosages on cytokine in LPS-induced mouse macrophage RAW 264.7 were determined. The result indicated UWE of the three stages had no inhibitory effect on pro-inflammatory cytokine TNF-α and PGE2. Moreover, UWE could significantly increase TNF-α. On the contrary, UWE of the three stages significantly reduced pro-inflammatory cytokine IL-6. Among the extracts UWE-stage 2 also had the highest inhibition up to 85.2 % at 125 ppm level. UWE of the three stages also decreased anti-inflammatory cytokine IL-10 with increase of their dosages. Among them UWE-stage 2 had the most anti-inflammatory potential by decreasing the ratio of IL-6/IL-10. The ratio decreased from 4.22 to 2.97. Finally, Western blot was applied to analyze the effects of UWE from daylily growth stages 2~4 with different dosage on inflammatory enzyme in LPS-induced mouse macrophage RAW 264.7. The result showed UWE-stage 2 reduced NO production by inhibiting iNOS expression.
