- 學位論文
黑鳳梨及乳酸菌發酵黑鳳梨之機能性探討及對小鼠巨噬細胞RAW 264.7的抗發炎作用之評估
Functional investigation of black pineapple and lactic acid bacteria fermented black pineapple, and evaluation of their anti-inflammatory effects on RAW 264.7 cells
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摘要
鳳梨為台灣盛產的水果之一 (產季3~7月)，但鳳梨的水分含量高，不易長期儲存且容易褐變腐敗，市面上已有多種延長鳳梨儲存期限的方法；例如製作成果乾、果醬、蜜餞等產品，其中台灣又以金鑽鳳梨 (台農17號) 為大宗，為了解決鳳梨有時生產過剩或者滯銷等問題，除了上述之處理方式之外，本研究欲利用金鑽鳳梨開發保健食品素材，因其價格相對一些具有機能性之特殊食材較便宜，若將鳳梨進行陳化製作成黑鳳梨，不僅能解決上述問題亦能為保健食品產業多一項新選擇，實驗中進行糖度、pH值、彩度等物性分析，並篩選出最適合在鳳梨中生長的乳酸菌，將乳酸菌擴大培養並對鳳梨及黑鳳梨進行乳酸發酵，觀察乳酸菌於鳳梨及黑鳳梨中的生長狀況及發酵效率。並且初步分析鳳梨、黑鳳梨及乳酸發酵液中的生物活性、抗氧化能力及抗菌活性，結果顯示黑鳳梨及乳酸菌發酵黑鳳梨具有較佳的效果，後續實驗以黑鳳梨及乳酸菌發酵黑鳳梨進行RAW 264.7巨噬細胞中脂多醣 (LPS) 誘導的抗發炎作用和抑制分子機制，RAW 264.7 細胞以LPS刺激細胞發炎，用不同濃度的黑鳳梨 (BP)、乳酸菌發酵黑鳳梨 (FBP) 培養，通過Griess試驗測定NO含量及使用ELISA kit測定促發炎激素含量，包括IL-6、IL-1β和TNF-α及抗發炎激素IL-10，結果顯示，NO含量隨著樣品濃度提高呈依賴性下降， BP及FBP以預防及治療模式皆可抑制IL-6產生，對於IL-1β促發炎激素而言，黑鳳梨主要通過預防模式發揮抑制作用，而乳酸菌發酵黑鳳梨則通過治療模式發揮抑制作用。TNF-α促發炎激素方面，無論BP或FBP皆是以治療模式在抑制激素的產生，而IL-10抗發炎激素之分泌量，BP及FBP皆藉由治療模式促進細胞激素的產生。ROS活性氧化物之含量也可藉由BP及FBP抑制產生，進一步確立抗發炎之能力。綜合整體表現，黑鳳梨和乳酸菌發酵黑鳳梨極具有潛力成為保健食品素材，未來於產品開發上有著不錯的前景。
並列摘要
Pineapple is one of the abundant fruits produced in Taiwan, and its season is from March to July. However, due to its high-water content, pineapples are not easy to store for long periods and are prone to browning and spoilage. Various methods are available on the market to extend the shelf life of pineapples, such as making dried fruit, jam, and candied products. Among these, Taiwan mainly produces the Golden Diamond Pineapple (Tainung No. 17). To address the issues of occasional overproduction or sluggish sales of pineapples, this study aims to develop a functional food ingredient using the Golden Diamond Pineapple. Compared to some functional specialty ingredients, pineapples are relatively cheaper. If the pineapples are aged to produce black pineapples, this not only resolves the aforementioned issues but also offers a new option for the functional food industry. In the experiments, physical properties such as sugar content, pH value, and color were analyzed, and the most suitable lactic acid bacteria for growth in pineapples were selected. These bacteria were cultured and used for lactic acid fermentation of both pineapples and black pineapples, with the growth and fermentation efficiency of the bacteria being observed. Initial analyses of the bioactivity, antioxidant capacity, and antibacterial activity in pineapples, black pineapples, and their fermentation liquids were conducted. Results indicated that black pineapples and lactic acid-fermented black pineapples exhibited better effects. Subsequent experiments focused on the anti-inflammatory effects and inhibitory mechanisms of black pineapples and lactic acid-fermented black pineapples on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophages. The RAW 264.7 cells were stimulated with LPS to induce inflammation and then cultured with varying concentrations of black pineapples (BP) and lactic acid-fermented black pineapples (FBP). The NO content was measured using the Griess assay, and pro-inflammatory cytokine levels, including IL-6, IL-1β, TNF-α, and the anti-inflammatory cytokine IL-10, were determined using ELISA kits. The results showed that the NO content decreased in a dose-dependent manner with increasing sample concentrations. Both BP and FBP could inhibit IL-6 production in both prevention and therapy models. For the pro-inflammatory cytokine IL-1β, black pineapples primarily exhibited inhibitory effects in the prevention model, while lactic acid-fermented black pineapples showed inhibitory effects in the therapy model. Regarding the pro-inflammatory cytokine TNF-α, both BP and FBP inhibited cytokine production in the therapy model. For the anti-inflammatory cytokine IL-10, both BP and FBP promoted cytokine production in the therapy model. The levels of reactive oxygen species (ROS) were also inhibited by BP and FBP, further establishing their anti-inflammatory capabilities. Overall, black pineapples and lactic acid-fermented black pineapples show great potential as functional food ingredients, with promising prospects for future product development.
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