- 學位論文
小金英萃取物在脂多醣刺激RAW264.7巨噬細胞之抗發炎潛力
Anti-inflammatory potential of Ixeris chinensis extract in lipopolysaccharide-stimulated RAW264.7 macrophages
摘要
發炎反應為我們人體受到外來病原體感染所引起的抵禦機制，發炎反應亦被發現與許多疾病相關。使用類固醇是治療慢性發炎最有效的抗發炎藥物，而長期使用類固醇藥物做為抗炎藥物所帶來副作用是許多人關注的問題。因此，從中草藥萃取具有抗發炎作用開始受到藥物研究與開發的重視。小金英（Ixeris Chinensis），為菊科（Compositae）苦蕒菜屬（Ixeris），是在臺灣常見的傳統中草藥，傳統上常用於解熱、治療消炎與各種疾病。 本實驗初步證實小金英水萃取物與小金英有機溶劑萃取物具有潛在抗炎之潛力。在本實驗中，以LPS刺激RAW264.7小鼠巨噬細胞作為發炎反應之細胞模式。ICWE對細胞存活力之影響，數據顯示ICWE對RAW 264.7巨噬細胞顯示低毒性（2500 μg/mL）。ICWE通過Mouse Inflammation Antibody Array檢測抗發炎潛力，再透過RT-PCR評估促炎細胞因子，如IL-1β、IL-6與COX-2等mRNA的表達，結果顯示ICWE可以抑制促炎細胞因子的基因表現。此外，ICWE抗發炎活性藉由檢測一氧化氮（NO）與反應性氧氣物質（ROS）生產，本研究顯示ICWE在100μg/ mL顯著抑制一氧化氮與反應性氧氣物質的產生。而在西方墨點法的實驗結果顯示ICWE可以抑制磷酸化的MAPK與磷酸化NF-κB蛋白質的表現，說明ICWE會影響MAPK與NF-κB的活化。有機溶劑萃取之小金英藉由檢測一氧化氮的產生初步判斷抗發炎之潛力。以上的實驗結果顯示，ICWE發揮顯著的抑制發炎反應之效果，可望在將來被開發做為天然的抗發炎藥物。
並列摘要
Inflammation is the body's response to injury, has been found to be associated with many diseases. Treatments using drugs such as corticosteroids have been proven effective, but many people concern about the side effects by prolonged use of corticosteroids as anti-inflammatory regimens. Accordingly, herbal extracts with anti-inflammatory activity began to be valued for pharmaceutical research and development. Ixeris Chinensis is species of Compositae and belongs to the genus Ixeris. Perennial herb commonly found in Taiwan. Traditionally used to relieve fever, anti-inflammatory and treating various ailments. We preliminarily evaluated the water extract of Ixeris chinensis (ICWE) and Organic Solvent extracts of Ixeris chinensis with potential anti-inflammatory activity. In the current study, we used lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophages as the inflammatory cell model. The effect of ICWE on the cell viability, data showed that ICWE show low toxicity on RAW 264.7 macrophages cells (2500 μg/mL). Anti-inflammatory potential of ICWE evaluated by Mouse Inflammation Antibody Array, and analysis of the interleukin-1β (IL-1β), Interleukin 6 (IL-6) and COX-2 proinflammatory cytokines mRNA expressions inflammatory cytokines by RT-PCR. The data showed that ICWE significant inhibits proinflammatory mRNA expressions. Furthermore, The anti-inflammatory activity of ICWE was determined by the inhibition of nitric oxide (NO) and reactive oxygen species (ROS) production. Results showed that ICWE significant inhibits NO and ROS at 100 μg/mL. In western blot experiment, the resulting showed that ICWE inhibits phosphorylation of MAPK and phosphorylation of NF-κB proteins, implying that ICWE can also inhibit of MAPK and NF-κB activation. The potential anti-inflammatory activity of Organic Solvent extracts of Ixeris chinensis was initially judged by the inhibition of NO production. These results suggest that ICWE holds the potential of being developed as anti-inflammatory herb drugs in the future.