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標題: 維生素與礦物質對雜交種吳郭魚巨噬細胞和周邊血液單核球免疫反應的影響
Effects of Vitamins and Minerals on the Immune Responses of Hybrid Tilapia (Oreochromis niloticus Oreochromis Mossambicus) Macrophages and Peripheral Blood Monocytes
作者: 洪紹文
Hung, Shao-Wen
關鍵字: In vivo;體內;In vitro;Macrophage;Minerals;Tilapia;Vitamins;體外;巨噬細胞;礦物質;吳郭魚;維生素
出版社: 獸醫學系暨研究所
引用: Abate A, Yang G, Dennery PA, Oberle S, Schroder H. 2000. Synergistic inhibition of cyclooxygenase-2 expression by vitamin E and aspirin. Free Radic Biol Med 29: 1135
本實驗著重於體外與體內實驗來探討單獨添加或合併添加抗氧化維生素 (維生素A、C及E) 和/或礦物質 (硒、鋅、銅、錳及鐵) 對雜交種吳郭魚 (Oreochromis niloticus × Oreochromis mossambicus) 週邊血液單核球來源、頭腎來源及脾臟來源之巨噬細胞的免疫功能影響。在體外試驗方面,結果發現適量的抗氧化維生素與礦物質會增加雜交種吳郭魚巨噬細胞增生能力和溶素酶的活性。然而,在合併添加高劑量的維生素 (A + C + E;每種維生素濃度為300 μg/mL) 或單獨添加礦物質 (硒、鋅、銅、錳或鐵,每種礦物質濃度為200、800或1,000 μg/mL) 則結果顯示反而會降低雜交種吳郭魚巨噬細胞增生能力和溶素酶的活性。合併添加上述所提到之任兩種維生素 (每種維生素濃度為100 μg/mL),則結果指出可以延長雜交種吳郭魚巨噬細胞增生能力直到添加維生素後72小時。但是,當單獨添加100 μg/mL維生素或40 μg/mL礦物質後,在雜交種吳郭魚巨噬細胞吞噬能力上並無顯著差異。另外,在感染前或感染後添加維生素或礦物質,結果發現皆可以降低雜交種吳郭魚巨噬細胞的死亡率,尤其當合併添加40或80 μg/mL礦物質和100 μg/mL維生素具有較大的保護能力。低劑量的維生素 (20-100 μg/mL vitamin A和C與20-40 μg/mL vitamin E) 會增加雜交種吳郭魚巨噬細胞一氧化氮的產生,同時會降低超氧化物的生成。但高劑量維生素 (超過100 μg/mL vitamin A和C與40 μg/mL vitamin E) 則會降低雜交種吳郭魚巨噬細胞的一氧化氮和超氧化物之生成。此外,結果顯示礦物質 (40、80、200、800或1,000 μg/mL) 亦會降低一氧化氮的生成。當給予濃度大於1,000或800 μg/mL 之維生素或礦物質時,雜交種吳郭魚巨噬細胞則會產生apoptosis或壞死。總結所有結果發現適當濃度的維生素或礦物質會增加雜交種吳郭魚巨噬細胞的免疫力,然而高濃度之維生素和礦物質則會導致細胞死亡。並且,在飼料中添加適當劑量之維生素和/或礦物質會增加雜交種吳郭魚巨噬細胞增生能力和保護力、維持巨噬細胞的活性、增加個體體重和體長及增加溶素酶活性。相反地,在飼料中若添加不適當劑量的維生素或礦物質和不適當合併之維生素和礦物質則會降低雜交種吳郭魚巨噬細胞超氧化物和一氧化氮的產生。因此,在飼料中添加適當劑量且適當合併給予維生素和礦物質是會增加雜交種吳郭魚巨噬細胞的免疫力。

The aim of this study was to determine the in vitro and in vivo effects of singular or combined the anti-oxidative vitamins (A, C, and E) and/or minerals (Se, Zn, Cu, Mn, and Fe) on the immune functions of hybrid tilapia, Oreochromis niloticus × Oreochromis mossambicus, peripheral blood monocyte-derived, anterior kidney-derived, and spleen-derived macrophages. Results from our in vitro study indicated that an optimal dose of vitamins and minerals increased cell proliferation and lysozyme activity. On the other hand, the above activities decreased at the high doses of combined vitamins (A + C + E group, each 300 μg/mL) or singular minerals (Se, Zn, Cu, Mn, and Fe groups, each 200, 800 or 1,000 μg/mL). Interestingly, we found that combining two of the aforementioned vitamins (A + C, A + E, and C + E groups, each 100 μg/mL) was able to prolong cell proliferative time up to 72 h compared with singular vitamin addition. The results of the phagocytosis assay showed that there was no difference between 100 μg/mL vitamin-treated or 40 μg/mL minerals-treated macrophages. Before or after adding vitamins or minerals during infection, addition of vitamins decreased the percentage of dead cells and a greater effect was observed for mineral (each 40 or 80 μg/mL) and vitamin (each 100 μg/mL) combinations. At the same time, we found that a low dose of vitamins increased nitric oxide production and decrease superoxide production, but high dose of vitamins decreased superoxide and nitric oxide production. Furthermore, minerals also decreased nitric oxide production at concentrations of 40, 80, 200, 800 or 1,000 μg/mL. The threshold concentrations for cell death by necrosis and/or apoptosis were more than 1,000 and 800 μg/mL for vitamins and minerals, respectively. In conclusion, appropriate concentration of vitamins or minerals can increase tilapia macrophage immunity; nevertheless, extreme concentrations of vitamins or minerals are lethal to cells. In the in vivo study, an optimal dose of vitamins and/or minerals in diets increased macrophage proliferation and protective activity, maintained macrophage viability, increased body weight and length, and increased lysozyme activity; however, at improper doses and combinations of vitamins or minerals a decrease was observed. Furthermore, vitamins and/or minerals at any doses and combinations in diets decreased superoxide and nitric oxide production. Therefore, appropriate doses and combinations of vitamins and/or minerals in diets may increase tilapia macrophages immunity.
其他識別: U0005-2706200701143200
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