本研究目的在評估茶樹籽萃取物為非皂素與兒茶素之類黃酮 (NCF) 對於改善因腫瘤壞死因子 (tumor necrosis factor-α，TNF-α) 所誘導的肝細胞胰島素抗性與患高血壓之糖尿病老鼠的高血壓與血糖改善效果。
首先，我們在人類肝癌細胞 Hep G2培養液中，分別加入 5μM 胰島素、5μM 胰島素及 30 ng / ml TNF-α，經2~6小時觀察。結果顯示TNF-α會降低IR、 IRS-1、 IRS-2、 PI3K-α、 Akt / PKB、 GLUT-2、 AMPK、 GCK、丙酮酸激酶和 PPAR-γ等蛋白質表現；並活化了 p65、 MAPKs （p38、 JNK1/2 和 ERK1/2）、 iNOS 及 COX-2 等；這顯示 TNF-α 使 Hep G2 細胞表現出胰島素抗性。在已產生胰島素抗性的Hep G2 細胞中加入不同濃度 NCF （500、 1000、 2000 ppm），發現Hep G2 細胞不僅胰島素信號傳導的蛋白質表達增加，而且也降低了發炎蛋白，如 p38，JNK1/2， iNOS 和 COX-2。我們推測 NCF 可藉著抑制p38、 JNK1/2、 iNOS 和 COX-2 蛋白質表現達到改善 TNF-α 誘導的胰島素抗性。
第二，我們進行 NCF 對於患高血壓之糖尿病老鼠的高血壓與血糖改善實驗。8週齡高血壓老鼠 (SHR) 分為 4 組 (每組 8 隻)：(1) RD 組:常規飲食。(2) DM組:注射streptozotocin (STZ, 50 mg/kg) 和 nicotinamide (NA, 60 mg/kg)，並餵食高脂飲食（HFD）。(3) NCF 0.5 組別與 (4) NCF 5 組別 : 注射STZ / NA餵食HFD並分別加口服灌餵NCF 0.5 mg/kg/day、NCF 5 mg/kg/day，持續餵食四週後。老鼠的第二型糖尿病被成功的誘發出而造成體重減輕、高血壓、高血糖、高糖化血色素、低胰島素、低腸促胰激素、高的 IL-6、 TNF-α、 MDA 及惡化了血管收縮與舒張的蛋白調節 (AGTR1, AGTR2, ETB, eNOS) 及調降了骨骼肌與肝臟葡萄糖代謝蛋白 (AMPK, IRB, IRS-1, PI3K, Akt/PKB, PPAR-γ, GLUT-4, HKase, PKase, GSK-3α, PEPCK, G6Pase) 與肝臟的自由基清除蛋白 (catalase, GPx)。上述的高血糖、高血壓、高血脂等相關生化指標、葡萄糖代謝、血壓調控與抗氧化反應皆可被 NCF改善並呈現劑量依賴性地改善。
由本次的研究結果，我們認為NCF具有對抗第二型糖尿、抗過氧化和抗發炎作用。雖然 NCF 並不含有皂素與兒茶素但擁有高量的可以調控第二型糖尿病的總黃酮 (40.9 μg/mg)。因此由本研究證實從茶籽萃取物中分離出的NCF具有發展成為控制第二型糖尿病並改善糖尿病引起之血管併發症的藥劑潛力。

The aim of this study is to investigate the extract of Camellia sinensis (L.) O. Kuntze seeds, contained non-catechin flavonoids (NCF), in improving hepatocytes insulin resistance by TNF-α and ameliorating hypertension and hyperglycemia on SHR with T2DM.
First, we used Hep G2 cell to be treated with 5μM insulin or with 5μM insulin plus 30 ηg/ml TNF-α for 2 to 6 hours. TNF-α decreased the protein expressions of IR, IRS-1, IRS-2, PI3K-α, Akt/PKB, GLUT-2, AMPK, GCK, pyruvate kinase, and PPAR- γ, and activated p65 and MAPKs (p38, JNK1/2 and ERK1/2), iNOS and COX-2. It showed that TNF-α induced the insulin resistance of Hep G2 cells. The Hep G2 cells with insulin resistance treated with NCF (500, 1000, 2000 ppm) not only increased the protein expressions of insulin signaling, but also reduced the inflammation proteins, as p38, JNK1/2, iNOS and COX-2. We suggest that NCF could ameliorate TNF-α induced insulin resistance through inhibiting p38, JNK1/2, iNOS and COX-2.
Second, we examined the anti-hypertension and anti-hyperglycemia effects of NCF treating in SHR with T2DM. 8-week-old SHRs were divided into four groups (Every group contains 8 rats): (1) RD group, fed regular diet, (2) DM group, which were injected with streptozotocin (STZ, 50 mg/kg) and nicotinamide (NA, 60 mg/kg) and fed high-fat-diet (HFD), (3) NCF 0.5 group and (4) NCF 5 group, which were injected with STZ/NA and fed HFD plus oral gavage NCF 0.5 mg/kg/day, and NCF 5 mg/kg/day for four weeks respectively. T2DM was successfully induced as evidenced by lower body weight; and higher blood pressure, blood glucose, HbA1c; and lower insulin, GLP-1; and higher IL-6, TNF-α, MDA; and deterioration of proteins regulating vasocontraction and vasodilation (AGTR1, AGTR2, ETB, eNOS) in vascular vessel, regulating glucose metabolism (AMPK, IRB, IRS-1, PI3K, Akt/PKB, PPAR-γ, GLUT-4, HKase, PKase, GSK-3α, PEPCK, G6Pase) in skeletal muscle and liver and free radicals scavengers (catalase, GPx) in liver. These biochemical parameters were improved dose dependently by NCF。
From the present results, we think that NCF had anti-T2DM, anti-peroxidation and anti-inflammatory effects. NCF possessed high concentrations of total flavonoid (40.9 μg/mg), not saponins and catechins, could efficiently control T2DM. Thus, NCF could potentiality be developed into an agent for the control of T2DM and related vascular complication.

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