近來許多研究顯示，血漿同半胱胺酸（homocysteine, Hcy）稍微升高是造成心血管疾病（cardiovasular disease, CVD）的獨立危險因子。人類血漿Hcy正常範圍為5-15 mM，大於15 mM即為同半胱胺酸血症（hyperhomocysteinemia, HHcy）。本研究室近年發現，國人CVD患者血漿Hcy濃度顯著高於正常人（12.2±0.3 vs. 8.5±0.4 mM）。然而，HHcy導致CVD的機制未明，HHcy促進血管內活性氧生成所產生的氧化壓力導致低密度脂蛋白（low density lipoprotein）及內皮細胞（endothelial cell）膜脂質過氧化可能為主要原因。此外，過氧化現象亦會促進血小板吸附（adhesion）能力及血小板活性進而造成血栓症。故本研究利用餵食甲硫胺酸(methionine)誘發大白鼠HHcy後，餵以山藥（Dioscores）來探討其對HHcy及氧化壓力之影響。餵以出生六週的大白鼠（Sprague-Dawley）食用Met（1 g/Kg body wt./day）十週誘發HHcy。將HHcy大白鼠隨機分為五組：Met 組、Met＋vitamin C組（200 mg/Kg body wt./day）及三個Met＋山藥組（D1、D2及D3）。山藥劑量分別為1、2.5及5 g/Kg body wt./day。分別於食用山藥及vitamin C八週與十二週時抽血並犧牲老鼠。血漿以7-Fluorobenzo-2-oxa-1,3-diazole-4-sulfonic acid （SBD-F）衍生化後，利用高效能液相層析儀（HPLC）分析血漿Hcy濃度。以血小板凝集儀（aggregometer）偵測thrombin（3.2 U/ml）誘發性血小板凝集反應。利用thiobarbituric acid reactive substances (TBARS)反應，以malondialdehyde（MDA）表示脂質過氧化。利用luminal當探針，以化學冷光偵測器（chemiluminescent）分析肝臟ROS含量。利用酵素動力學分析肝臟中抗氧化酵素。肝臟glutathione（GSH）和 Glutathion disulfide（GSSG）以O-Phthalaldehyde衍生化後，利用螢光分光光度計（Fluorescence spectrophotometer）分析。
結果發現， Met fed十週後，血漿Hcy濃度由6.7±1.0 mM顯著增加至 21.7±4.5 mM。食用三個劑量山藥八~十二週後，血漿Hcy濃度顯著比未食山藥（Met）組低，且與控制組相近。食用中及高劑量山藥十二週後，血小板凝集反應顯著比未食山藥（Met）組小。食用三個劑量山藥十二週後，也使血漿MDA濃度及肝臟ROS含量顯著比未食山藥（Met）組低，並與控制組相似。當大白鼠食用中及高劑量山藥十二週後，肝臟之catalase活性顯著比未食山藥（Met）組高。然而，食用山藥對肝臟GSH及GSSG沒有顯著影響。
本實驗顯示，食用山藥可以恢復因Met所誘發的HHcy。此外，HHcy可藉由攝取山藥而減少血小板凝集、脂質過氧化及氧化壓力。本研究結論，山藥對高蛋白飲食（高Met攝取），可能誘發的HHcy與氧化壓力導致的相關疾病具有保護功能，值得推廣山藥成為保健食品。

Recently, a moderately elevated plasma homocystein (Hcy) level has been identified as an independent risk factor for cardiovascular disease (CVD). Normal range of Hcy levels is 5-15 mM in human plasma. Plasma concentration of Hcy > 15 mM is hyperhomocysteinemia (HHcy). In this laboratory, we found that patients with CVD had significantly higher plasma Hcy concentration than normal person (12.2 ± 0.3 vs. 8.5± 0.4 mM). However, the mechanism by which HHcy leads to CVD is not known. HHcy-induced oxidative stress due to the production of reactive oxygen species (ROS) resulted in oxidation of low density lipoprotein (LDL) and endothelial membrane may contribute to the possible mechanism. In addition, peroxidation in cell membrane could activate platelet and promote adhesion leading to thrombosis. Therefore, the purpose of this study was to investigate the antioxidative effects of Dioscores in methionine (Met)-induced HHcy animals. HHcy was induced by oral feeding Met (1 g/Kg body wt./day) for 10 weeks in Sprague-Dawley rats at the age of 6 weeks. HHcy rats were randomly divided into five groups: Met group without Dioscorea feeding, Met＋vitamin C group supplemented with vitamin C 200 mg/Kg body wt./day, and 3 Dioscores groups (D1, D2 and D3) supplemented with freeze-dried Dioscorea powder 1, 2.5 and 5 g/Kg body wt./day, respectively. Animals were sacrificed after 8 or 12 weeks of Dioscorea feeding. Plasma levels of Hcy were assayed based on HPLC of the fluorescent 7-Fluorobenzo-2-oxa-1,3-diazole-4-sulfonic acid (SBD-F) derivative. Platelet aggregation induced by thrombin (3.2 U/ml) was analyzed with an aggregometer. Plasma lipid peroxidation was measured as thiobarbituric acid reactive substances (TBARS) expressed in malondialdehyde (MDA) equivalents. ROS generation was monitored with luminometer using luminal as the probe. Activities of antioxidative enzymes were measured by kinetics of enzymes using UV spectrophotometer. Hepatic concentrations of glutathione and glutathion disulfide were quantified by fluorescence spectrophotometer after derivatized with O-Phthalaldehyde.
The results of this study showed that plasma Hcy levels were significantly increased from 6.7±1.0 to 21.7±4.5 mM after 10 weeks of Met feeding. Eight and 12 weeks after Dioscorea feeding, plasma Hcy of D1, D2, and D3 were significantly lower than that of Met, and similar to the basal level prior to Met feeding. Twelve weeks after Dioscorea feeding, thrombin-induced platelet aggregation of D2 and D3 were significantly lower than that of Met group. Plasma MDA levels and hepatic ROS of D1, D2, and D3 were similar to that of Ctl group and significantly lower than that of Met group after 12 weeks of Dioscorea feeding. The activities of hepatic catalase in D2 and D3 groups were significantly elevated compared to Met group at 12 weeks. However, feeding Dioscorea 8 or 12 weeks did not significantly change hepatic levels of GSH and GSSG.
Results of our study indicated that HHcy induced by Met could be reversed by Dioscorea feeding. In addition, Dioscorea feeding could alleviate platelet aggregation, lipid peroxidation, and oxidative stress in HHcy. The protective effects of Dioscores feeding in oxidative stress induced by high protein (Met) intake. Suggested Dioscores as a valuable functional food.

目 錄
頁數
中文摘要………………………………………………………… Ⅰ
英文摘要………………………………………………………… Ⅳ
致謝……………………………………………………………… Ⅶ
目錄……………………………………………………………… Ⅷ
表目錄…………………………………………………………… Ⅸ
圖目錄…………………………………………………………… Ⅹ
縮寫符號………………………………………………………..ⅩⅢ
第一章、 前言…………………………………………………… 1
第二章、 文獻探討……………………………………………… 4
第三章、 實驗材料及方法……………………………………… 16
第四章、 結果…………………………………………………… 36
第五章、 討論…………………………………………………… 47
參考文獻………………………………………………………… 57
表次……………………………………………………………… 69
圖次……………………………………………………………… 80
自述………………………………………………………………100
表 目 錄
頁數
表一、實驗動物飼料組成……………………………………… 69
表二、食用Methionine十週大白鼠的體重變化量…………… 71
表三、食用山藥八週大白鼠的體重變化量……………………. 72
表四、食用山藥十二週大白鼠的體重變化量…………………. 73
表五、不同山藥劑量與不同時間處理對變異數之影響………. 74
表六、不同山藥劑量處理對大白鼠血漿生化值及肝臟
酵素活性之比較…………………………………………. 75
表七、不同時間處理對大白鼠血漿生化值及肝臟酵素
活性之比較………………………………………….…… 75
表八、食用山藥八、十二週對大白鼠肝臟中
GSH/GSSG比率…………………………………………. 76
表九、Hcy與血漿生化值及肝臟酵素活性的相關性…………. 77
表十、Platelet Aggregation與血漿生化值及肝臟酵素
活性的相關性……………………………...……………. 78
表十一、MDA與血漿生化值及肝臟酵素活性的相關性……… 78
表十二、ROS與血漿生化值及肝臟酵素活性的相關性……… 79
圖 目 錄
頁數
圖一、Homocysteine代謝圖……………...……………….…… 80
圖二、抗氧化酵素作用……………...…………………….…… 81
圖三、Homocysteine造成動脈硬化機制…………………..…… 82
圖四、血漿Homocysteine層析圖譜...…………………….…… 83
圖五、血漿Vit. C層析圖譜………...…………………….…..… 83
圖六、食用Methionine十週前後大白鼠血漿中Hcy濃度…..… 84
圖七、食用山藥八週後大白鼠血漿中Hcy濃度……….………. 84
圖八、食用山藥十二週後大白鼠血漿中Hcy濃度…….………. 85
圖九、不同山藥劑量處理第八週及第十二週大白鼠
血漿Hcy濃度………………………...………………… 86
圖十、食用山藥八週後，大白鼠thrombin誘發性
血小板凝集反應………………………………………… 86
圖十一、食用山藥十二週後，大白鼠thrombin誘發性
血小板凝集反應…………………………………...…. 87
圖十二、不同山藥劑量處理第八週及第十二週大白鼠
thrombin誘發性血小板凝集反應…...……………….. 87
頁數
圖十三、食用山藥八週後，大白鼠血漿中MDA含量………… 88
圖十四、食用山藥十二週後，大白鼠血漿中MDA含量……… 88
圖十五、不同山藥劑量處理第八週及第十二週大白鼠
血漿中MDA含量………………………………….…. 89
圖十六、食用山藥八週後，大白鼠肝臟中ROS含量…………… 89
圖十七、食用山藥十二週後，大白鼠肝臟中ROS含量…..…… 90
圖十八、不同山藥劑量處理第八週及第十二週大白鼠
肝臟中ROS含量………………………..………….…. 90
圖十九、食用山藥八週後，大白鼠血漿中Vit. C濃度…….…. 91
圖二十、食用山藥十二週後，大白鼠血漿中Vit. C濃度…….…. 91
圖二十一、食用山藥八週後，大白鼠肝臟中SOD活性….……. 92
圖二十二、食用山藥十二週後，大白鼠肝臟中SOD活性….…. 92
圖二十三、食用山藥八週後，大白鼠肝臟中Catalase活性.…. 93
圖二十四、食用山藥十二週後，大白鼠肝臟中Catalase活性…. 93
圖二十五、不同山藥劑量處理第八週及第十二週
大白鼠肝臟中Catalase活性...…..…………….…… 94
圖二十六、食用山藥八週後，大白鼠肝臟中
Glutathione Peroxidase活性………………………. 94
頁數
圖二十七、食用山藥十二週後，大白鼠肝臟中
Glutathione Peroxidase活性………...…..………… 95
圖二十八、不同山藥劑量處理第八週及第十二週
大白鼠肝臟中Glutathione Peroxidase活性………. 95
圖二十九、食用山藥八週後，大白鼠肝臟中
Glutathione reductase活性……………..……..……. 96
圖三十、食用山藥十二週後，大白鼠肝臟中
Glutathione reductase活性……………………...…….. 96
圖三十一、食用山藥八週後，大白鼠肝臟中
Glutathione濃度.…………………………………… 97
圖三十二、食用山藥八週後，大白鼠肝臟中
Glutathione disulfide濃度……...…………………… 97
圖三十三、食用山藥十二週後，大白鼠肝臟中
Glutathione濃度…..…………………………...…… 98
圖三十四、食用山藥十二週後，大白鼠肝臟中
Glutathione disulfide濃度……...…………………… 98
圖三十五、不同山藥劑量處理第八週及第十二週
大白鼠肝臟中Glutathione 濃度…………………… 99

Aebi, H. Catalase. In: Methods of Enzymatic Analysis (Bergmeyer HV, ed) 1983 pp. 637. Chemie , Weinheim, FRG.
Araghiniknam M, Chung S, Nelson-White T, Eskelson C, Watson RR. Antioxidant activity of dioscorea and dehydroepiandrosterone (DHEA) in older humans. Life Sci. 1996; 59: PL147-157.
Atamna H, Ginsburg H. The malaria parasite supplies glutathione to its host cell--investigation of glutathione transport and metabolism in human erythrocytes infected with Plasmodium falciparum. Eur J Biochem. 1997; 250: 670-679.
Aw TY, Wierzbicka G, Jones DP. Oral glutathione increases tissue glutathione in vivo. Chem Biol Interact. 1991; 80: 89-97.
Barroso MP, Gomez-Diaz C, Lopez-Lluch G, Malagon MM, Crane FL, Navas P. Ascorbate and alpha-tocopherol prevent apoptosis induced by serum removal independent of Bcl-2. Archives of Biochemistry & Biophysics. 1997; 343: 243-248.
Barroso G, Morshedi M, Oehninger S. Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. Hum Reprod. 2000; 15: 1338-1344.
Bellamy MF, McDowell IF, Ramsey MW, Brownlee M, Bones C, Newcombe RG, Lewis MJ. Hyperhomocysteinemia after an oral methionine load acutely impairs endothelial function in healthy adults. Circulation. 1998; 98: 1848-1852.
Bellomo G, Mirabelli F, DiMonte D, Richelmi P, Thor H, Orrenius C, Orrenius S. Formation and reduction of glutathione-protein mixed disulfides during oxidative stress. A study with isolated hepatocytes and menadione (2-methyl-1,4-naphthoquinone). Biochemical Pharmacology. 1987; 36: 1313-1320.
Benzuly KH, Padgett RC, Kaul S, Piegors DJ, Armstrong ML, Heistad DD. Functional improvement precedes structural regression of atherosclerosis. Circulation. 1994; 89: 1810-1818.
Blom HJ, Kleinveld HA, Boers GH, Demacker PN, Hak-Lemmers HL, Te Poele-Pothoff MT, Trijbels JM. Lipid peroxidation and susceptibility of low-density lipoprotein to in vitro oxidation in hyperhomocysteinaemia. Eur J Clin Invest. 1995; 25: 149-154.
Blundell G, Jones BG, Rose FA, Tudball N. Homocysteine mediated endothelial cell toxicity and its amelioration. Atherosclerosis. 1996; 122: 163-172.
Bridges AB, Scott NA, McNeill GP, Pringle TH, Belch JJ. Circadian variation of white blood cell aggregation and free radical indices in men with ischaemic heart disease. European Heart Journal. 1992; 13: 1632-1636.
Bunout D, Garrido A, Suazo M, Kauffman R, Venegas P, de la Maza P, Petermann M, Hirsch S. Effects of supplementation with folic acid and antioxidant vitamins on homocysteine levels and LDL oxidation in coronary patients. Nutrition. 2000; 16: 107-110.
Cavalca V, Cighetti G, Bamonti F, Loaldi A, Bortone L, Novembrino C, De Franceschi M, Belardinelli R, Guazzi MD. Oxidative stress and homocysteine in coronary artery disease. Clin Chem. 2001; 47: 887-892.
Cavallini D, De Marco C, Dupre S, Rotilio G. The copper catalyzed oxidation of cysteine to cystine. Arch Biochem Biophys. 1969; 130: 354-361.
Celermajer DS, Sorensen K, Ryalls M, Robinson J, Thomas O, Leonard JV, Deanfield JE. Impaired endothelial function occurs in the systemic arteries of children with homozygous homocystinuria but not in their heterozygous parents. J Am Coll Cardiol. 1993; 22: 854-858.
Cerutti PA. Prooxidant states and tumor promotion. Science. 1985; 227: 375-381.
Chambers JC, McGregor A, Jean-Marie J, Obeid OA, Kooner JS. Demonstration of rapid onset vascular endothelial dysfunction after hyperhomocysteinemia: an effect reversible with vitamin C therapy. Circulation. 1999; 99: 1156-1160.
Chang S-J, Chang C-N, Hsu J-C, Lee Y-S, Shen C-H. Homocysteine, vitamin B6 and lipid profile in cardovascular diseas. Nutrition. 2002(in press).
Clarke R, Daly L, Robinson K, Naughten E, Cahalane S, Fowler B, Graham I. Hyperhomocysteinemia: an independent risk factor for vascular disease. N Eng J Med. 1991; 324: 1149-1155.
Dardik R, Varon D, Tamarin I, Zivelin A, Salomon O, Shenkman B, Savion N. Homocysteine and oxidized low density lipoprotein enhanced platelet adhesion to endothelial cells under flow conditions: distinct mechanisms of thrombogenic modulation. Thromb Haemost. 2000; 83: 338-344.
den Heijer M, Blom HJ, Gerrits WB, Rosendaal FR, Haak HL, Wijermans PW, Bos GM. Is hyperhomocysteinaemia a risk factor for recurrent venous thrombosis? Lancet. 1995; 345: 882-885.
Domagala TB, Libura M, Szczeklik A. Hyperhomocysteinemia following oral methionine load is associated with increased lipid peroxidation. Thromb Res. 1997; 87: 411-416.
Draper HH, Hadley M. Malondialdehyde determination as index of lipid peroxidation. Methods in Enzymology. 1990; 186: 421-431.
Dudman NP, Hicks C, Wang J, Wilcken DE. Human arterial endothelial cell detachment in vitro: its promotion by homocysteine and cysteine. Atherosclerosis. 1991; 91: 77-83.
Dudman NP, Guo XW, Crooks R, Xie L, Silberberg JS. Assay of plasma homocysteine: light sensitivity of the fluorescent 7-benzo-2-oxa-1, 3-diazole-4-sulfonic acid derivative, and use of appropriate calibrators. Clinical Chemistry. 1996; 42: 2028-2032.
Dumaswala UJ, Wilson MJ, Jose T, Daleke DL. Effect of a glycerol-containing hypotonic medium on erythrocyte phospholipid asymmetry and aminophospholipid transport during storage. Biochim Biophys Acta. 1997; 1330: 265-273.
Dumaswala UJ, Zhuo L, Jacobsen DW, Jain SK, Sukalski KA. Protein and lipid oxidation of banked human erythrocytes: role of glutathione. Free Radic Biol Med. 1999; 27: 1041-1049.
Dumaswala UJ, Zhuo L, Mahajan S, Nair PN, Shertzer HG, Dibello P, Jacobsen DW. Glutathione protects chemokine-scavenging and antioxidative defense functions in human RBCs. Am J Physiol Cell Physiol. 2001; 280: C867-C873.
Durak I, Karabacak HI, Buyukkocak S, Cimen MY, Kacmaz M, Omeroglu E, Ozturk HS. Impaired antioxidant defense system in the kidney tissues from rabbits treated with cyclosporine. Protective effects of vitamins E and C. Nephron. 1998; 78: 207-211.
Durand P, Lussier-Cacan S, Blache D. Acute methionine load-induced hyperhomocysteinemia enhances platelet aggregation, thromboxane biosynthesis, and macrophage-derived tissue factor activity in rats. FASEB. 1997; 11: 1157-1168.
Dykman KD, Tone C, Ford C, Dykman RA. The effects of nutritional supplements on the symptoms of fibromyalgia and chronic fatigue syndrome. Integr Physiol Behav Sci. 1998; 33: 61-71.
Frank L, Massaro D. Oxygen toxicity. Am J Med. 1980; 69: 117-126.
Frei B. Reactive oxygen species and antioxidant vitamins: mechanisms of action. Am J Med. 1994; 97(3A): 5S-13S.
Frei B. On the role of vitamin C and other antioxidants in atherogenesis and vascular dysfunction. Experimental Biology & Medicine. 1999; 222: 196-204.
Fryer RH, Wilson BD, Gubler DB, Fitzgerald LA, Rodgers GM. Homocysteine, a risk factor for premature vascular disease and thrombosis, induces tissue factor activity in endothelial cells. Arterioscler Thromb. 1993; 13: 1327-1333.
Guan Y, Zhao S. Yishou jiangzhi (de-blood-lipid) tablets in the treatment of hyperlipemia. J Tradit Chin Med. 1995; 15: 178-179.
Hagen TM, Aw TY, Jones DP. Glutathione uptake and protection against oxidative injury in isolated kidney cells. Kidney Int. 1988; 34: 74-81.
Harker LA, Slichter SJ, Scott CR, Ross R. Homocystinemia. Vascular injury and arterial thrombosis. N Engl J Med. 1974; 291: 537-543.
Harker LA, Ross R, Slichter SJ, Scott CR. Homocystine-induced arteriosclerosis. The role of endothelial cell injury and platelet response in its genesis. J Clin Invest. 1976; 58: 731-741.
Harker LA. Harlan JM. Ross R. Effect of sulfinpyrazone on homocysteine-induced endothelial injury and arteriosclerosis in baboons. Circulation Research. 1983; 53: 731-739.
Harpel PC, Chang VT, Borth W. Homocysteine and other sulfhydryl compounds enhance the binding of lipoprotein(a) to fibrin: a potential biochemical link between thrombosis, atherogenesis, and sulfhydryl compound metabolism. Proc Natl Acad Sci USA. 1992; 89: 10193-10197.
Hajjar KA. Homocysteine-induced modulation of tissue plasminogen activator binding to its endothelial cell membrane receptor. J Clin Invest. 1993; 91: 2873-2879.
Heinecke JW. Superoxide-mediated oxidation of low density lipoprotein by thiols. In: Cerruti PA, Fridovich I, McCord JM, eds. Oxy-radicals in molecular biology and pathology. New York: Alan R Liss. 1988: 443-457.
Hissin PJ, Hilf R. A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal Biochem. 1976; 74: 214-226.
Hladovec J. Methionine, pyridoxine and endothelial lesion in rats. Blood Vessels 1980; 17: 104-109.
Holvoet P, Perez G, Zhao Z, Brouwers E, Bernar H, Collen D. Malondialdehyde-modified low density lipoproteins in patients with atherosclerotic disease. J Clin Invest. 1995; 95: 2611-2619.
Holvoet P. Oxidative modification of low-density lipoproteins in atherothrombosis. Acta Cardiol. 1998; 53: 253-260.
Hu k, Dong A, Yao X, Kobayashi H, Iwasaki S. Antineoplastic agents; I. Three spirostanol glycosides from rhizomes of Dioscorea collettii var. hypoglauca. Planta Medica. 1996; 62: 573-575.
Iwu MM, Okunji CO, Ohiaeri GO, Akah P, Corley D, Tempesta MS. Hypoglycaemic activity of dioscoretine from tubers of Dioscorea dumetorum in normal and alloxan diabetic rabbits. Planta Med. 1990; 56: 264-267.
Jakubowski H. Metabolism of homocysteine thiolactone in human cell cultures. Possible mechanism for pathological consequences of elevated homocysteine levels. J Biol Chem. 1997; 272: 1935-1942.
Kanani PM, Sinkey CA, Browning RL, Allaman M, Knapp HR, Haynes WG. Role of oxidant stress in endothelial dysfunction produced by experimental hyperhomocyst(e)inemia in humans. Circulation. 1999; 100: 1161-1168.
Keizer HG, Pinedo HM, Schuurhuis GJ, Joenje H. Doxorubicin (adriamycin): a critical review of free radical-dependent mechanisms of cytotoxicity. Pharmacol Ther. 1990; 47: 219-231.
Kosower NS, Kosower EM. The glutathione status of cells. Int Rev Cytol. 1978; 54: 109-160.
Kuller LH, Evans RW. Homocysteine, vitamins, and cardiovascular disease. Circulation. 1998; 98: 196-199.
Kushleika J, Checkoway H, Woods JS, Moon JD, Smith-Weller T, Franklin GM, Swanson PD. Selegiline and lymphocyte superoxide dismutase activities in Parkinson''s disease. Ann Neurol. 1996; 39: 378-381.
Lang D, Kredan MB, Moat SJ, Hussain SA, Powell CA, Bellamy MF, Powers HJ, Lewis MJ. Homocysteine-induced inhibition of endothelium-dependent relaxation in rabbit aorta: role for superoxide anions. Arteriosclerosis Thrombosis & Vascular Biology. 2000; 20: 422-427.
Lee ME, Wang H. Homocysteine and hypomethylation: a novel link to vascular disease. Trends Cardiovasc Med. 1999; 9: 49-54.
Lentz SR, Sadler JE. Inhibition of thrombomodulin surface expression and protein C activation by the thrombogenic agent homocysteine. Journal of Clinical Investigation. 1991; 88: 1906-1914.
Lentz SR, Sobey CG, Piegors DJ, Bhopatkar MY, Faraci FM, Malinow MR, Heistad DD. Vascular dysfunction in monkeys with diet-induced hyperhomocyst(e)inemia. J Clin Invest. 1996; 98: 24-29.
Li XP. Experimental study on anti-senility of the 4 famous Chinese herbs produced in Huaiqing area. Zhong Xi Yi Jie He Za Zhi. 1991; 11: 486-487.
Ling CD, Zhang ZJ, Chen ZL. Studies on nutritional effects of traditional Chinese tonics with strengthened nutrients on pregnant women and rats. Zhongguo Zhong Xi Yi Jie He Za Zhi. 1996; 16: 270-273.
Loscalzo J. The oxidant stress of hyperhomocyst(e)inemia. J Clin Invest. 1996; 98: 5-7.
Malinow MR, Kang SS, Taylor LM, Wong PW, Coull B, Inahara T, Mukerjee D, Sexton G, Upson B. Prevalence of hyperhomocyst(e)inemia in patients with peripheral arterial occlusive disease. Circulation. 1989; 79: 1180-1188.
Mano T, Sinohara R, Sawai Y, Oda N, Nishida Y, Mokumo T, Asano K, Ito Y, Kotake M, Hamada M. Changes in lipid peroxidation and free radical scavengers in the brain of hyper- and hypothyroid aged rats. J Endocrinol 1995; 147:361-365.
Marklund S, Marklund G. Involvenment of superoxide anion radical in autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 1974; 47: 469-474.
Matthias D, Becker CH, Riezler R, Kindling PH. Homocysteine induced arteriosclerosis-like alterations of the aorta in normotensive and hypertensive rats following application of high doses of methionine. Atherosclerosis. 1996; 122: 202-216.
McCully KS. Vascular pathology of homocysteinemia: Implications for the pathogenesis of arteriosclerosis. Am J Pathol. 1969; 56: 111-128.
McCully KS, Wilson RB. Homocysteine theory of arteriosclerosis. Atherosclerosis. 1975; 22: 215-227.
McCully KS. Homocysteine theory of arteriosclerosis: Development and current ststus. Atherosclerosis Reviews. 1983; 11: 157, Gotto AM Jr. Pauletti R (eds.), New York: Raven Press.
McCully KS, Vezeridis MP. Histopathological effects of homocysteine thiolactone on epithelial and stromal tissues. Experimental & Molecular Pathology. 1989; 51: 159-170.
McDonald L, Bray C, Field C, Love F, Davies B. Homocystinuria thrombosis and the blood-platelets. Lancet. 1964; 1: 745-746.
Mehta J, Li D, Mehta JL. Vitamins C and E prolong time to arterial thrombosis in rats. Journal of Nutrition. 1999; 129: 109-112.
Moat SJ, Bonham JR, Cragg RA, Powers HJ. Elevated plasma homocysteine elicits an increase in antioxidant enzyme activity. Free Radic Res. 2000; 32: 171-179.
Mohamed HE, El-Swefy SE, Hagar HH. The protective effect of glutathione administration on adriamycin-induced acute cardiac toxicity in rats. Pharmacol Res. 2000; 42: 115-121.
Mori N, Hirayama K. Long-term consumption of a methionine-supplemented diet increases iron and lipid peroxide levels in rat liver. J Nutr. 2000; 130: 2349-2355.
Munday R. Toxicity of thiols and disulphides: involvement of free-radical species. Free Radic Biol Med. 1989; 7: 659-673.
Nakazono K, Watanabe N, Matsuno K, Sasaki J, Sato T, Inoue M. Does superoxide underlie the pathogenesis of hypertension? Proc Natl Acad Sci USA 1991; 88:10045-10048.
Nappo F, De RN, Marfella R, De LD, Ingrosso D, Perna AF, Farzati B, giugliano D. Impairment of endothelial functions by acute hyperhomocysteinemia and reversal by antioxidant vitamins. JAMA. 1999; 281: 2113-2118.
Nath KA, Fischereder M, Hostetter TH. The role of oxidants in progressive renal injury. Kidney Int. 1994; 45 (suppl. 45): S111-S115.
Nightingale AK, James PP, Morris-Thurgood J, Harrold F, Tong R, Jackson SK, Cockcroft JR, Frenneaux MP. Evidence against oxidative stress as mechanism of endothelial dysfunction in methionine loading model. Am J Physiol Heart Circ Physiol. 2001; 280: H1334-H1339.
Nygard O, Nordrehaug JE, Refsum H, Ueland PM, Farstad M, Vollset SE. Plasma homocysteine levels and mortality in patients with coronary artery disease. N Engl J Med. 1997; 337: 230-236.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry. 1979; 95: 351-358.
Ohtsuki T, Matsumto M, Suzuki K, Taniguchi N, Kamada T. Mitrochondrial lipid peroxidation and superoxide dismutase in rat hypertensive target organs. Am J Physiol. 1995; 268: H1418-1421.
Outinen PA, Sood SK, Liaw PC, Sarge KD, Maeda N, Hirsh J, Ribau J, Podor TJ, Weitz JI, Austin RC. Characterization of the stress-inducing effects of homocysteine. Biochem J. 1998; 332: 213-221.
Ozaki M, Fuchinous S, Teraoka S, Ota K. The in vivo cytoprotection of ascorbic acid against ischemial reoxygenation injury of rat liver. Arch Biochem Biophys. 1995; 318: 439-445.
Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 1967; 70: 158-169.
Pascoe GA, Fariss MW, Olafsdottir K, Reed DJ. A role of vitamin E in protection against cell injury. Maintenance of intracellular glutathione precursors and biosynthesis. Eur J Biochem. 1987; 166: 241-247.
Pickett WC, Cohen P. Mechanism of the thrombin-mediated burst in oxygen consumption by human platelets. J Biol Chem. 1976; 251: 2536-2538.
Pietrzik K, Bronstrup A. Folate in preventive medicine: a new role in cardiovascular disease, neural tube defects and cancer. Ann Nutr Metab. 1997; 41: 331-343.
Podmore ID, Griffiths HR, Herbert KE, Mistry N, Mistry P, Lunec J. Vitamin C exhibits pro-oxidant properties. Nature. 1998; 392: 559.
Powell CV, Nash AA, Powers HJ, Primhak RA. Antioxidant status in asthma. Pediatr Plumonol. 1994; 18:34-38.
Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990’s. Nature. 1993; 362: 801-809.
Saldeen T, Li D, Mehta JL. Differential effects of alpha-and gamma-tocopherol on low-density lipoprotein oxidation, superoxide activity, plateler aggregation and arterial thrombogenesis. J Am Coll Cardiol. 1999; 34: 1208-1215.
“SAS Users Guide.” SAS Institute Inc., P. O. Box 10066, Raleigh, N.C. 27605, 1979.
Selhub J, Bagley LC, Miller J, Rosenberg IH. B vitamins, homocysteine, and neurocognitivte function in the elderly. Am J Clin Nutr. 2000; 71: 614S-620S.
Sheu JR, Lee CR, Hsiao G, Hung WC, Lee YM, Chen YC, Yen MH. Comparison of the relative activities of alpha-tocopherol and PMC on platelet aggregation and antioxidative activity. Life Sciences. 1999; 65: 197-206.
Shi MM, Kugelman A, Iwamoto T, Lian L, Forman HJ. Quinone-induced oxidative stress elevates glutathione and γ-glutamylcysteine synthetase activity in rat lung epithelial L2 cells. J Biol Chem. 1994; 269: 26512-26517.
Smith P, Krohn RI, Hermanson GT, Mallia A K, Gartner F H, Prorenzano MD, Fujimoto EK, Goeke NM, Olson BT. Klenk, DC. Measurement of protein using bicinchonic acid. Anal Biochem. 1985; 15: 76-85.
Stamler JS, Osborne JA, Jaraki O, Rabbani LE, Mullins M, Singel D, Loscalzo J. Adverse vascular effects of homocysteine are modulated by endothelium-dervied relaxing factor and related oxides of nitrogen. J Clin Invest 1993; 91:308-318.
Stamler JS, Slivka A. Biological chemistry of thiols in the vasculature and in vascular-related disease. Nutr Rev. 1996; 54: 1-30.
Starkebaum G, Harlan JM. Endothelial cell injury due to copper-catalyzed hydrogen peroxide generation from homocysteine. J Clin Invest. 1986; 77: 1370-1376.
Steiner M, Anastasi J. Vitamin E. An inhibitor of the platelet release reaction. J Clin Invest. 1976; 57: 732-737.
Tawakol A, Omland T, Gerhard M, Wu JT, Creager MA. Hyperhomocyst(e)inemia is associated with impaired endothelium-dependent vasodilation in humans. Circulation. 1997; 95: 1119-1121.
Toborek M, Kopieczna-Grzebieniak E, Drozdz M, Wieczorek M. Increased lipid peroxidation and antioxidant activity in methioine-induced hepatitis in rabbits. Nutrition. 1996; 12: 543-537.
Ueland PM, Refsum H. Plasma homocysteine, a risk factor for vascular disease: plasma levels in health, disease and drug therapy. J Lab Clin Med. 1989; 114: 473-501.
Ungvari Z, Pacher P, Rischak K, Szollar L, Koller A. Dysfunction of nitric oxide mediation in isolated rat arterioles with methionine diet-induced hyperhomocysteinemia. Arterioscler Thromb Vasc Biol. 1999; 19: 1899-1904.
Ungvari Z, Starkadi-Nagy E, Bagi Z, Szollar L, Koller A. Simultaneously increased TxA2 activity in isolated arterioles and platelets of rats with hyperhomocysteinemia. Arterioscler Thromb Vasc Biol. 2000; 20: 1203-1208.
Upchurch GR Jr, Welch GN, Fabian AJ, Freedman JE, Johnson JL, Keaney JF Jr, Loscalzo J. Homocyst(e)ine decreases bioavailable nitric oxide by a mechanism involving glutathione peroxidase. J Biol Chem. 1997; 272: 17012-17107.
Upchurch GR Jr, Welch GN, Fabian AJ, Pigazzi A, Keaney JF Jr, Loscalzo J. Stimulation of endothelial nitric oxide production by homocyst(e)ine. Atherosclerosis. 1997; 132: 177-185.
Ventura P, Panini R, Verlato C, Scarpetta G, Salvioli G. Peroxidation indices and total antioxidant capacity in plasma during hyperhomocysteinemia induced by methionine oral loading. Metabolism. 2000; 49: 225-228.
Verhoef P, Kok FJ, Kruyssen DA, Schouten EG, Witteman JC, Grobbee DE, Ueland PM, Refsum H. Plasma total homocysteine, B vitamins, and risk of coronary atherosclerosis. Arterioscler Thromb Vasc Biol. 1997; 17: 989-995.
Voutilainen S, Morrow JD, Roberts LJ, Alfthan G, Alho H, Nyyssonen K, Salonen JT. Enhanced in vivo lipid peroxidation at elevated plasma total homocysteine levels. Arterioscler Thromb 1999; 19:1263-1266.
Wall RT, Harlan JM, Harker LA, Striker GE. Homocysteine-induced endothelial cell injury in vitro: a model for the study of vascular injury. Thromb Res. 1980; 18: 113-121.
Welch GN, Upchurch GR Jr, Keaney Jr JF, Loscalzo J. Homocyst(e)ine decreases cell redox potential in vascular smooth muscle cells. J Am Coll. 1996; 27: 164A.
White JG, Rao GH, Gerrard JM. Effects of nitroblue tetrazolium and vitamin E on platelet ultrastructure, aggregation, and secretion. Am J Pathol. 1977; 88: 387-402.
White CR, Brock TA, Chang LY, Crapo J, Briscoe P, Ku D, Bradley WA, Gianturco SH, Gore J, Freeman BA. Superoxide and peroxynitrite in atherosclerosis. Proc Natl Acad Sci USA. 1994; 91: 1044-1048.
Xu D, Neville R, Finkel T. Homocysteine accelerates endothelial cell senescence. FEBS Lett. 2000; 470: 20-24.
Yagi K. Lipid peroxides and human diseases. Chem Phys Lipids. 1987; 45: 337-351.
Zhang F, Slungaard A, Vercellotti GM, Iadecola C. Superoxide-dependent cerebrovascular effects of homocysteine. Am J Physiol. 1998; 274: R1704-1711.
劉新裕、王昭月、宋麗梅、高介治。山藥優良品系70W04之選育、栽培與品質研究。中華農業研究。1993. 42. pp. 265-279。
徐輝妃、黃鵬。山藥的營養及保健價值與實用法介紹。花蓮區農業專訊。1997. pp.7-9。
馮鶯。山藥澄粉藥膳療法延緩慢性腎功能不全進程的研究。中華護理雜誌。1997. pp. 707-709。
潘文涵、章雅惠、陳正義、吳幸娟、曾明淑、高美丁。以二十四小時飲食回顧法評估國人膳食營養狀況。國民營養現況，1993－1996國民營養健康狀況變遷調查表（修訂版）。行政院衛生署，1998. pp. 29-51。
張素瓊、范慧如。維生素B6營養狀況調查。國民營養現況，1993－1996國民營養健康狀況變遷調查表（修訂版）。行政院衛生署，1998. pp. 195-203。
林璧鳳、林榮富、葉文婷、潘文涵。1993-1993年國人之葉酸營養狀況。國民營養現況，1993－1996國民營養健康狀況變遷調查表（修訂版）。行政院衛生署，1998. pp. 217-227。
賴宏亮、鄧秀玫、劉新裕，許博文。山藥藥材鑑別之研究。藥材食品分析，1999; 7(7): 313-325。
劉新裕、張同吳、林義恭、陳淑芬、王昭月、朱戩良、王順成。山藥之品種特性、生產潛力淺力、物化性質與抗氧化研究。中華農業研究。1999. 48. pp. 1-22。
張素瓊, 李懿純, 廖瓊瑤, 蕭莉茹. 2000.1.20-1.21.維生素B群缺乏下甲硫胺酸對血壓,血中同半胱胺酸濃度及血小板凝集之影響。中國生物學會八十九年年會Abstract book : 39。