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  • 學位論文

以脂多醣誘發RAW264.7小鼠巨噬細胞發炎模式探討台灣產南美假櫻桃乙醇萃取物的抗發炎功效

Study on Anti-inflammatory Effect of Ethanol Extract from Jamaica Cherry (Muntingia calabura Linn.) Fruits Gathered in Taiwan Estimated with LPS-Stimulated RAW264.7 Macrophage Cells

指導教授 : 楊登傑 林昭田

摘要


南美假櫻桃 (Muntingia calabura Linn.),屬於文丁科(Muntingiaceae)西印度櫻桃屬(Muntingia)之植物。原產於美洲熱帶,常分佈於台灣南部低海拔地區,目前在台灣大多作為行道樹和庭園樹,其果實可食用。在我們的預實驗中發現南美假櫻桃的乙醇萃取物有良好的抗氧化活性。因此,本研究再進一步利用管柱層析獲得抗氧化活性更高的南美假櫻桃乙醇萃取物之區分物 (50%丙酮+50%甲醇),並利用脂多醣 (lipopolysaccharide, LPS)誘發RAW264.7細胞株之發炎模式,探討南美假櫻桃乙醇萃取物之區分物的抗發炎活性及其可能之機轉。實驗結果發現,南美假櫻桃乙醇萃取物之區分物能有效降低一氧化氮合成酶 (inducible nitric oxide synthase, iNOS)的蛋白表現量,進而減少一氧化氮 (nitric oxide, NO)的生成量,也抑制促發炎細胞激素【腫瘤壞死因子-α (tumor necrosis factor-α, TNF-α)、介白素-6 (interleukin-6, IL-6)、介白素-1β (interleukins-1β, IL-1β)】的生成。南美假櫻桃的抗發炎調控機轉為:(1) 調控轉錄因子 NF-κB (nuclear factor kappa B )途徑:抑制inhibitor to κB kinase α/β (IKKα/β)的活化,並抑制NF-κB p65及p50的蛋白表現;(2) 調控有絲裂原活化蛋白質激酶 mitogen-activated protein kinases (MAPKs):抑制p38及c-Jun NH2-terminal kinase 2 (JNK2)的活化;(3) 調控janus kinase (JAK)/signal transducer and activator of transcription (STAT)途徑:抑制JAK2的活化,進而抑制STAT1及STAT3的活化;(4)調控nuclear factor erythroid-2-related factor 2 (Nrf2) / heme oxygenase-1 (HO-1)途徑:增加Nrf2的蛋白表現量,進而增加HO-1的蛋白表現量。

關鍵字

南美假櫻桃 RAW264.7 NF-κB MAPK JAK/STAT Nrf2/HO-1

並列摘要


Jamaica cherry (Muntingia calabura Linn.) belonging to Muntingiaceae family nativly grows in tropical America. It also grows in low altitude areas in southern Taiwan and is mostly planted as shade and contryyard tree. Its furit can be eaten directly. Our pre-experiment found that the ethanol extract of Jamaica cherry fruit had a good antioxidant capacity. The fraction with better antioxidant capacity was further prepared from the ethanol extract through column chromatography with the elution of 50% acetone and 50% methanol. It was used to evaluate the anti-inflammatory effect with lipopolysaccharide (LPS)-stimulated RAW264.7 cell line in the investigation. Results show that the ethanol extract fraction could suppress the expression of inducible nitric oxide synthase (iNOS) protein, production of nitric oxide (NO) and secretion of proinflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). The attenuation of LPS-induced inflammatory responses was closely related to inhibition of the inhibitor to κB kinase (IKK) α/β activation and NF-κB p65 and p50 subunits protein expression (NF-κB pathway), down-regulation of phosphorylation of p38 and c-Jun NH2-terminal kinase 2 (JNK2) (MAPKs pathway), suppression of phosphorylation of Janus kinase 2 (JAK2), signal transducer and activator of transcription 1 (STAT1) and signal transducer and activator of transcription 3 (STAT3) (JAK/STAT pathway), and enhancement of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein prodution (Nrf2/HO-1 pathway).

參考文獻


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被引用紀錄


郭麗淳(2016)。以肝癌細胞 (HepG2) 探討臺灣產南美假櫻桃果實乙醇萃取物調控鎳誘導 VEGF 之表現〔碩士論文,中山醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0003-1209201619002500

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