臺灣博碩士論文加值系統

近年的研究顯示，低密度脂蛋白的氧化修飾扮演著啟動和促進動脈粥狀硬化和相關的心血管疾病的角色，能抑制低密度脂蛋白氧化之抗氧化劑可能減退動脈粥狀硬化之發展。低密度脂蛋白粒子的構型和其所含的多元不飽和脂肪酸及維生素E都是影響其在活體和離體情況下氧化敏感度的重要因素。目前已知一些源自活血化瘀的中藥和食物中的某些植物天然物可抑制低密度脂蛋白氧化。本研究探討烘烤黑豆對血膽固醇正常人低密度脂蛋白氧化敏感性之影響，包括三項：(ㄧ)比較青仁黑豆烘烤前與後抗氧化物質，諸如：MDA、花青素、genistein、維生素E、GSHPx和SOD之含量變化；(二)選定17名健康未抽煙成年男性，以日常飲食為基礎，每日給予40公克烘烤青仁黑豆(台南3號)為期3週，檢測服用黑豆前、後以及停後一個月體內血漿、低密度脂蛋白抗氧化物質之含量變化，包括維生素E、花青素、GSHPx、SOD及genistein；(三)檢測三階段低密度脂蛋白的氧化敏感性。結果顯示：離體試驗檢測青仁黑豆與烘烤黑豆的抗氧化功效顯示兩者均能抑制MDA生成量達84%以上。青仁黑豆於烘烤後其花青素的含量顯著降低(118.41 ±5.51 vs. 72.73 ±13.07 unit/g)(p<0.01); 烘烤青仁黑豆中genistein顯著高於未烘烤前之黑豆(3.61 ±0.26 vs. 0.63 ±0.10 μg/g)(p<0.01)；至於青仁黑豆於烘烤前後的維生素E及GSHPx含量皆未達顯著差異；而青仁黑豆於烘烤後其SOD之活性顯著降低(2345.0 ±663.92 vs. 555.0 ±400.34 unit/g)(p<0.001)。在活體內試驗顯示，服用烘烤黑豆3週後其血漿中TG顯著下降(175.27 ±18.28 vs. 135.96 ±50.87 mg/dL)(p<0.01)。且服用後顯著延長LDL氧化延遲期(95.18 ±21.94 vs. 118.88 ±34.30 min)(p=0.0075)。同時服用後與停止服用一個月相較呈現顯著差異。(118.88 ±34.30 vs. 97.35 ±24.81 min)(p<0.05）。受試者服用烘烤黑豆後其血漿中花青素含量顯著高於服用前(0.08 ±0.09 vs. 0.17 ±0.14 unit/mg plasma-chol)(p<0.05)。LDL中維生素E取對數值於服用前與服用後呈顯著差異(1.36 ±0.45 vs. 1.53 ±0.29 nmol/LDL-C)(p<0.05)；服用烘烤黑豆三週後，LDL之氧化延遲期與血漿中花青素呈顯著正相關(r=0.58813)(p<0.01)。但LDL中維生素E取對數值卻與LDL氧化延遲期呈負相關(r=-0.70915)(p﹦0.0014)。紅血球中SOD、GSHPx及LDL中genistein於三階段未見顯著差異。總而言之，補充烘烤黑豆顯著降低受試者血中TG值，同時延長LDL氧化延遲期(p<0.05)，且與血漿中anthocyanin值變化有關。

Many recent studies have demonstrated that oxidative modification of low density lipoprotein(LDL) play a key role in the development of atherosclerosis and coronary heart disease(CHD). Antioxidants capable of inhibiting LDL oxidation may reduce atherosclerosis. The structure and contents of polyunsaturated fatty acid(PUFA) and vitamin E in LDL are important factors affecting the oxidative susceptibility of LDL in vitro and vivo. Several antioxidants from traditional Chinese herbs on blood stasis and dietary phytochemicals capable of inhibiting LDL oxidation are reported. In this study, we investigated the effect of dietary supplementation with roasted black bean on susceptibility of LDL to oxidative modification. Seventeen healthy male subjects intake 40g of roasted black bean every day for 3 weeks. Compare the antioxidant contents of black beans before and after roasted; fasting blood was drawn and LDL was isolated and treated with 10mM Cu2+ for LDL lag time determination before, after and stop of the ingestion in vivo, with the plasma cholesterol, triglycerol and anthocyanin levels and the contents of vitamin E, genistein in LDL and the activity of GSHPx, SOD in RBC were also determined. In vitro modle test, organic extracts of green cotyledon of black beans(GBB) and roasted black beans(RBB) significantly inhibited MDA formation 84%. The contents of anthocyanin in RBB was significantly lower than that in GBB(118.4 ±5.51 unit/g and 72.73 ±13.07 unit/g). The contents of genistein in RBB significantly higher than that in GBB(3.61 ±0.26μg/g and 0.63 ±0.10μg/g). However, the content of vitamin E and GSHPx activity in RBC no significant difference. The activity of SOD in RBB was significantly lower than that in GBB(2345.0 ±663.92 unit/g, and 555.0 ±400.34 unit/g). In vivo modle test, the result show plasma triglycerol from 175.27±18.28 mg/mL to 135.96±50.87 mg/mL, decreasing significantly after roasted black bean ingestion. Lag time before LDL oxidation prolonged significantly from 95.18±21.94 min to 118.89±34.30 min in 3 weeks after ingestion. Taking 3 weeks roasted black beans significantly prolonged LDL oxidation from 118.88 ±34.30 min to 97.35 ±24.81 min in stop after 1 month ingestion. The anthocyanin content was significantly higher in plasma isolated before than that in after intaking RBB from 0.08 ±0.09 unit/mg to 0.17 ±0.14 unit/mg plasma-chol. The log of Vit E content significantly increased from 1.36 ±0.45 nmol/LDL-C in LDL before intaking RBB to 1.53 ±0.29 nmol/LDL-C in LDL after intaking. While lag phase of LDL in subjects supplemented with GBB was positively correlated with plasma anthocyanin (r=0.58813), it was negatively correlated with log vitamin E of LDL. The activities of SOD, GSHPx in RBC and the contents of genistein in LDL were no differences. In conclusion, intaking black bean significantly decreased plasma TG, and significantly prolonged the lag phase of LDL oxidation, which may be resulted from high content of anthocyanin in plasma.

表、圖目錄
縮寫表
中文摘要 1
英文摘要 3
第一章 前言 5
第二章 文獻回顧 7
第一節 活性氧與自由基 7
第二節 體內的抗氧化系統 9
第三節 黃豆的臨床研究 14
第四節黑豆功效與抗氧化研究 20
第三章 實驗設計及方法 23
第四章 實驗材料與方法 25
壹、受試者之篩選 25
貳、烘烤黑豆之處理 25
參、烘烤黑豆抗氧化物質含量之分析 25
一、烘烤黑豆麩胱甘過氧化活性之測定 26
二、烘烤黑豆超氧化歧化活性之測定 27
三、烘烤黑豆維生素E之含量測定 29
四、烘烤黑豆Genistein含量測定 31
五、烘烤黑豆花青素含量的測定 33
六、抑制Thiobarbituric acid reactive substance(TBARS)
生成之測量 33
肆、血液處理 35
伍、低密度脂蛋白之氧化敏感性測試 36
陸、生化分析 39
ㄧ、測定血漿總膽固醇濃度 39
二、測定血漿中三醯甘油酯之濃度 40
三、血漿中花青素含量之測定 40
四、紅血球麩胱甘過氧化活性之測定 41
五、紅血球超氧歧化活性之測定 42
六、測定LDL中所含之膽固醇量 42
七、LDL中維生素E含量之測定 43
八、LDL中Genistein含量之測定 43
柒、統計分析 44
第五章 結果 45
第六章 討論 51
第七章 結論 57
參考文獻 75
表目錄
表一 黑豆烘烤前與後其抗氧化物質含量之比較 59
表二 受試者之基本資料 60
表三 服用烘烤黑豆之健康受試者，於不同時期其體內抗氧化
物質含量 61
表四 健康受試者服用烘烤黑豆，於不同時期其體內抗氧化物
質含量與低密度脂蛋白氧化延遲期之相關性 62
表五 服用烘烤黑豆之邊緣高血脂症者，於不同時期其血脂質
及LDL氧化延遲期之變化 63
表六 服用烘烤黑豆之邊界高血脂症者，於不同時期其體內抗
氧化物質含量 64
表七 健康受試者24小時回憶法的營養素攝取量 65
表八 受試者的食物頻率 66
圖目錄
圖ㄧ 服用烘烤黑豆之健康受試者，在不同時期其體內血脂質
的變化 73
圖二 服用烘烤黑豆之健康受試者，在不同時期低密度脂蛋白
氧化延遲期之變化 74

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