評估不同陳化程度之黑檸檬水萃物抗氧化活性與其對小鼠巨噬細胞之抗發炎影響

Evaluation of the antioxidant properties of aqueous extract of black lemon (Citrus limon L. Brum. f.) at different aged periods and the anti-inflammatory effect in murine macrophages

葉科宏(Yeh Ke-Hung)

指導教授 ： 朱永麟

國立屏東科技大學/國際學院/食品科學國際碩士學位學程/碩士(2023年)

https://doi.org/10.6346/NPUST202300037

摘要

檸檬 (Citrus limon L. Burm. f.) 是台灣的一種重要的作物，其含有大量的類黃酮，已有多項研究證明能對於健康帶來益處，然而，在產季期間巨大的產量常造成市場滯銷的問題，因此，開發檸檬相關的多樣化產品是十分必要的。黑檸檬，又稱作陳年檸檬，通常是由檸檬製成的一種柑橘類乾果，經過微熱加工，直到呈現出黑褐色的外觀，並經過乾燥後保存。一般來說，炎症 (發炎) 是身體的免疫系統對刺激物的反應。腸道的炎症與一氧化氮 (NO) 的合成有著高度的相關性，這是由於誘導性一氧化氮合成酶 (iNOS) 上調的結果所導致。本研究將透過福林試劑 (Folin Ciocalteu) 比色法和氯化鋁比色法分別測定黑檸檬水萃物 (ABLE) 中的總酚含量和總黃酮含量，並通過鐵還原抗氧化能力 (FRAP) 來測定ABLE 的抗氧化能力，除此之外也測定了陳化過程中其總含糖量的變化。利用 Griess 反應測量由脂多醣 (LPS) 誘導的小鼠巨噬細胞 (RAW264.7) 釋放到培養基中的亞硝酸鹽之含量，在比較不同濃度之 ABLE 對於其抑制效果。對於抗炎之研究，通過西方墨點法(Western blotting) 進行對iNOS、COX-2、IL-6 以及 TNF-α 等促炎介質之評估。綜上所述，研究結果表明，ABLE 具有較高的總酚及總類黃酮含量，以及較佳的鐵環原能力，同時，在分析促炎因子蛋白的方面，也呈現出不錯的抑制效果，尤其是對 iNOS 蛋白的抑制效果有特別突出的表現，由此可知 ABLE 同時具有傑出的抗氧化以及抗發炎能力，未來能夠被應用於開發機能性食品原料之潛力。

關鍵字

陳化 ； 抗氧化能力 ； 黑檸檬 ； 抗發炎反應 ； 梅納反映

並列摘要

Lemon (Citrus limon L. Brum. f.) is an important crop in Taiwan. It contains a high amount of flavonoids, which have been reported for several health benefits. However, the abundant production usually causes sluggish market sales problems during the harvest period. Hence, it is necessary to develop diversified products of lemon. Black lemon (aged lemon) is a dried citrus fruit, normally made from lemon, which is processed through lightly thermal until it takes on the appearance of blackish-brown leather and becomes dry and preserved. Generally, inflammation is the body`s immune system's response to an irritant and could be classified into chronic and acute two major types of inflammation. Intestinal inflammation is highly correlated with the increased synthesis of nitric oxide (NO) as the consequence of up-regulated inducible nitric oxide synthase (iNOS). The study aimed to determine the total phenolic content and total flavonoid content in aqueous black lemon extract (ABLE) using Folin-Ciocalteu`s method and aluminum chloride colorimetric assay respectively, and the antioxidant capacity of ABLE was determined by FRAP (Ferric reducing antioxidant power). The effects of ABLE on
lipopolysaccharide (LPS) induced NO production in mouse macrophages
(RAW264.7) were investigated by measuring the amount of nitrite released into
the culture medium using the Griess reaction. For the anti-inflammatory effects,
pro-inflammatory mediators, including iNOS, tumor necrosis factor-α (TNF α), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), were examined by Western blotting. In summary, the result shows that ABLE has a high amount of total phenolic and flavonoid content, besides, it also shows great ferric-reducing ability, therefore, Besides, the results of the analysis of pro-inflammatory factor proteins also showed a great inhibitory effect, especially on the iNOS protein. Overall, based on the results we consider thatABLE might have outstanding antioxidant and anti-inflammation properties, in the future, it has the potential to be an ideal raw material for the development of functional food.

並列關鍵字

Aging process ； black lemon ； antioxidant ； anti-inflammatory ； Maillard reaction

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