第二型糖尿病的發生主因為胰島素阻抗 (Insulin resistance, IR)，而免疫系統失調造成發炎會加重 IR。自然殺手細胞 (Natural killer cells, NK) 為一種淋巴細胞，構成先天免疫系統之一環，可快速清除外來病毒及體內不正常細胞。在動物及人類之第 2 型糖尿病下觀察 NK 數量與活性均大幅受損，但 NK 在第 2 型糖尿病之角色目前尚不明確。本實驗室參考 NK-based 免疫療法，將 NK 注射至 Streptozotocin (STZ) 誘發之糖尿病鼠 BALB/c 與 C57BL/6，發現可改善 IR，然而其中機制與能否在體外大量培養 NK 仍待深入研究。因此，本實驗目的為以體外培養 NK，探討注射活化或死亡 NK (1×106 cells) 對於糖尿病鼠 BALB/c 之 IR及肝臟脂肪代謝之影響。結果顯示，添加 Interleukin-15 可大幅提高 NK 在體外培養的存活率。注射高劑量 NK 可顯著回復糖尿病鼠的葡萄糖耐受性、HOMA-IR 及 Glucose transporter type-4 (GLUT-4) 表現量。另外，給予高劑量 NK 注射亦提升肝臟脂肪代謝調控因子 peroxisome proliferator-activated receptors-α 表現量及降低脂質抗氧化。此外，BALB/c 鼠之血漿中促發炎細胞激素 Interleukin-6 會隨 NK 劑量增加而降低。但注射死亡 NK 者不明顯。綜合以上，注射以 Interleukin-2 及 Interleukin-15 誘導之 NK 可改善糖尿病小鼠模式下胰島素抗性和脂質氧化及發炎現象。

Insulin resistance (IR) plays a critical role in Type 2 diabetes mellitus (T2D) development. Immune cells dysfunction may exacerbate IR. The evidence revealed that natural killer cells (NK) were severely damaged in numbers and activity under diabetic condition. However, whether elevated NK numbers and activity contribute to IR is unclear. This study aims to investigate the effects of adoptive transfer NK on IR. The results revealed that exogenous interleukin-15 raised NK viability in vitro. In animal model, adoptive transfer NK (1 × 106 cells) improved glucose tolerance and IR and also the fat metabolic regulator expression in liver. Furthermore, adoptive transfer of dead NK (1 × 105 cells) did not restore glucose metabolism capacity suggesting that the mechanism may due to protein or cytokines released instead of ligands on NK surface. Adoptive transfer of NK also decreased interleukin-6 concentration in plasma, suggesting exogenous NK attenuated inflammation in diabetic mice. Collectively, adoptive transfer of NK improved insulin resistance and inflammation and liver fat oxidation in diabetic mice.

摘要 I
Abstract II
目錄 III
名詞縮寫表 V
圖目錄 VII
表目錄 VIII
附錄目錄 IX
壹、 緒論 1
貳、 文獻回顧 2
1. 糖尿病 2
1.1 第 1 型糖尿病 2
1.2 第 2 型糖尿病 3
1.3 胰島素阻抗 3
1.4 胰島素阻抗與脂肪代謝 3
1.5 糖尿病診斷標準 4
2. 自然殺手細胞 4
2.1 自然殺手細胞為基礎細胞免疫療法 5
2.2 糖尿病與自然殺手細胞 5
參、 實驗流程 9
1. 自然殺手細胞分離 9
2. 動物實驗 10
3. 實驗目的 11
肆、 實驗材料及方法 12
1. 實驗材料 12
1.1 動物 12
1.2 飼料 12
1.3 藥品 12
1.4 儀器設備 13
2. 實驗方法 15
2.1 細胞實驗 15
2.2 動物實驗 15
2.3 糖尿病相關分析 17
2.4 注射自然殺手細胞 18
2.5 分析樣品之取得 18
2.6 小鼠自然殺手細胞分選 19
2.7 細胞激素測定 20
2.8 自然殺手細胞表面標記分析 20
2.9 組織樣品蛋白質分析 20
3. 統計分析 23
伍、 結果 24
1. 細胞實驗部分 24
2. 動物實驗部分 25
陸、 討論 27
1. 細胞實驗部分 27
2. 動物實驗部分 27
3. 總結 30
柒、 參考文獻 31
捌、 圖表 36
玖、 附錄 53

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