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研究生： 江怡馨
Chiang, I-Hsin
論文名稱： 以動物和細胞模式來探討硫辛酸降低高血脂症之效應及機轉
Investigate the effects and possible mechanisms of lipoic acid in prevention of hyperlipidemia with animal and cell culture models
指導教授： 王應然
Wang, Ying-Jan
學位類別： 碩士
Master
系所名稱： 醫學院 - 環境醫學研究所
Department of Environmental and Occupational Health
論文出版年： 2010
畢業學年度： 98
語文別： 中文
論文頁數： 79
中文關鍵詞： 高脂血症 、硫辛酸 、倉鼠 、HMG-CoA reductase
外文關鍵詞： hypercholesterolemia, α-Lipoic acid, hamster, HMG-CoA reductase
相關次數： 點閱：187  下載：2 
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    • 由於國人飲食和生活習慣型態的改變，使得罹患高血脂症（hyperlipidemia）及其引發的心血管疾病（CVD）患者，都有逐年升高的趨勢。硫辛酸（LA）是細胞可自行少量產生的雙硫化合物，存於天然動植物中且具有相當強之抗氧化能力，而LA應用於氧化性壓力相關疾病的研究中，顯示 LA對於氧化性壓力造成的疾病有保護作用。本研究的目的是探討利用 LA對於倉鼠之降血脂及體內抗氧化能力的功效；以及 LA降低血脂的可能機制。實驗設計以雄性的倉鼠（Hamster）進行實驗，先餵養高膽固醇飼料 (額外添加膽固醇0.2%）三週，待倉鼠引起高血脂症後，再以腹腔注射方式給予LA每周三次持續十週。實驗結果發現LA可顯著降低高脂血症倉鼠的血清中總膽固醇（TC）、三酸甘油脂（TG）、低密度脂蛋白膽固醇（LDL-C）、高密度脂蛋白膽固醇（HDL-C）；並且可以顯著降低脂質累積在肝臟中的情形，改善膽固醇飲食中所造成的脂肪肝。也觀察到LA以及DHLA具有良好的抗氧化能力，提升了抗氧化酵素 GSH、GSH Px的活性，抑制脂質過氧化物的生成，進而降低罹患動脈粥狀硬化的風險，減緩動脈粥狀硬化發生的進展。在體外實驗中發現 LA可以有效降低 HMG-CoA reductase的活性，使得膽固醇在體內的濃度下降，以達到具有降低血脂的功效；與臨床治療高血脂症之用藥Statin類作用雷同。因此建議可以將 LA運用在臨床的治療或預防血脂異常症狀上。

    Hyperlipidemia is one of the major risk factors for the development of cardiovascular disease. There is accumulating data demonstrated hypercholesterolemia and oxidative stress play important roles in the development of atherosclerosis. LA has attracted considerable clinical interest as a cellular thiol-replenishing and redox-modulating agent. Biologically, LA functions as a cofactor of oxidative decarboxylation reactions in glucose metabolism to yield energy. It has been used for a long time in the western world to treat complications associated with diabetes. In the present study, we sought to determine whether LA is able to exert its antioxidant effect and alter blood lipid levels in hyperlipidemia’s animal modal. To induce hyperlipidemia, the male hamsters were given high cholesterol diet (contain 0.2% cholesterol) for 3 weeks. The hyperlipidemia animals were treated with or without intraperitoneal injections of LA three times per week for 10 weeks. We found that LA could decrease the serum levels of TC, TG, LDL-C, and HDL-C. LA markedly inhibited lipid accumulation in the liver. This findings suggested that LA would be a potent metabolic antioxidant source to quench free radicals by using ORAC assay and TBARS assay in vitro and in vivo. Furthermore, the activity of HMG-CoA reductase, the rate-limiting enzyme in cholesterol synthesis, were down-regulated by LA in vitro. Given its strong safety record, LA may have potential clinical applications for the treatment or prevention of hypercholesterolemia.

    目綠 第一章、序論 1 第二章、文獻回顧 2 第一節、代謝症候群與心血管疾病 2 第二節、血脂異常與心血管疾病 3 第三節、高血脂 5 一、血漿脂質 5 二、脂蛋白 6 三、膽固醇的代謝 8 四、脂肪肝的形成 9 五、高血脂動物模式 9 六、細胞模式 10 第四節、硫辛酸 12 一、硫辛酸的簡介 12 二、硫辛酸的治療潛能 13 第三章、研究目的 15 第四章、實驗設計 16 第一節、體內實驗 (In Vivo) 16 第二節、體外實驗 (In Vitro) 17 第五章、研究材料與方法 18 第一節、研究材料 18 第二節、體內實驗 (In Vivo)研究方法 21 第三節、體外實驗 (In Vitro)研究方法 31 第四節、統計分析 40 第六章、結果 41 第一節、體重、攝食量以及組織臟器重量變化 41 第二節、脂質代謝評估 41 第三節、肝臟組織觀察 43 第四節、倉鼠肝臟抗氧化功能評估 44 第五節、LA在HepG2細胞中的脂質評估 44 第六節、在HepG2細胞中LA對於HMG-CoA reductase表現的影響 45 第七節、LA對於HMG-CoA reductase活性的影響 45 第八節、LA、DHLA抑制LDL氧化評估 46 第七章、討論 48 第八章、結論與建議 53 參考文獻 54 圖表結果 60 附錄 78   圖目錄 Fig 1. Findings of livers outward appearances of male hamsters fed with high cholesterol diet for 13 weeks. 66 Fig 2. Liver were obtained from mice fed a normal diet or high-fat diet with LA or without LA. Representative photomicrographs of H&E staining in liver . 67 Fig 3. Liver were obtained from mice fed a normal diet or high-fat diet with or without LA . Representative pictures of oil red O staining in liver . 68 Fig 4. Antioxidant activities of LA in hyperlipidemia’s animal modal. 69 Fig 5. Effect of LA of oxidative stress on hepatic level in hamsters fed high-fat diet. 70 Fig 6. Effect of LA on cell viability of HepG2 cells. 71 Fig 7. Dose-dependent effects of LA on intracellular total cholesterol concentrations in HepG2 cells. 72 Fig 8. Effect of LA on Oil-Red O retention in HepG2 cells. 73 Fig 9. Effect of LA on the protein expression of HMG-CoA reductase in HepG2 cell. 74 Fig 10. Effect of LA on the mRNA expression of HMG-CoA reductase in HepG2 cell. 75 Fig 11. Effect of LA (A) and DHLA (B) on the diene production during CuSO4-induced lipid peroxidation of LDL. 77 Fig 12. Effect of LA (A) and DHLA (B) on the CuSO4-induced lipid peroxidation in LDL. 78 表目綠 文獻回顧 Table 1 Plasma lipid profiles and epidemiology of atherosclerotic diseases in Taiwan- a unique experience 4 圖表結果 Table 1. Effect of diet contained different dose LA on body weight, weight gain and food inake in male hamsters for 13 weeks. 61 Table 2. Effect of diet contained different dose LA on liver weight and Kidney weight in male hamsters for 13 weeks. 62 Table 3. Effect of diet contained different dose LA on serum lipid levels and atherogenic index in male hamsters for 13 weeks. 63 Table 4. Effect of diet contained different dose LA on liver total cholesterol and triglyceride in male hamsters for 13 weeks. 64 Table 5. Suppression of HMG CoA reductase activity by LA and DHLA. 75

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