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研究生:歐欣泙
研究生(外文):Hsin-ping Ou
論文名稱:MonacolinK之熱降解動力學分析與紅麴發酵產物保健功效研究
論文名稱(外文):Investigations on Monacolin K Thermal Degradation Kinetics Analysis and Healthy Effects of Monascus-fermented Products
指導教授:王俊權王俊權引用關係
指導教授(外文):Chiun-chuang Wang
學位類別:博士
校院名稱:靜宜大學
系所名稱:食品營養研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2007/07/
畢業學年度:95
語文別:英文
論文頁數:130
中文關鍵詞:紅麴熱降解動力抗氧化膽固醇尿酸倉鼠SAMP8老化學習記憶
外文關鍵詞:Antioxidant activityCholesterolUric acidHamster ratsSAMP8 miceAgingLearning and memoryMonacolin KThermal degradation kinetics
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紅麴菌已被證明可經由培養在米粒上,產生許多對人體有益的代謝產物。這些發酵產物包括一次代謝產物(如不飽和脂肪酸、酯類)以及二次代謝產物(色素、抗腐敗菌物質、膽固醇合成抑制劑(monacolin K)、降血壓物質及其他尚待鑑定之活性物質)。紅麴發酵產物已如同膳食補充品般被廣泛使用,並且應用在新產品開發中,做為一食品添加材料。雖然國外已有紅麴相關功能性的報告,然而主要著重於血壓及血糖的調節。本研究的目的在探討紅麴代謝產物之抗氧化能力、monacolin K之熱降解動力分析及調節倉鼠血脂質功效,同時更進一步評估其對延緩及改善老化促進小鼠學習記憶能力之效益。
紅麴發酵產物在配製之不同酸鹼性溶液(pH3-9)中,其monacolin K具有良好安定性。紅麴發酵產物之鹼性溶液(pH 9)經121℃加熱90min後,其膽固醇合成抑制物含量驟減至14.1%。經熱反應動力分析結果發現,膽固醇合成抑制物之熱降解屬於一次動力反應(r2=0.99)。含3%紅麴發酵產物溶液經90-121℃加熱處理,其膽固醇合成抑制物之熱降解反應的活化能、熱焓及熵值分別為126.64、 120.78 及0.22kJ/mol。紅麴發酵產物經不同乙醇濃度(50, 75, 100%)萃取所得之紅麴萃取物,皆對於DPPH自由基具有清除能力,其中以75%乙醇之紅麴萃取物效果顯著最佳,其清除自由基50%有效濃度(IC50)為6.67 ± 0.75 mg/ml。而在還原力之試驗結果顯示,其還原力皆隨樣品添加劑量的增加而增加。此外,在總多酚類化合物含量亦以75%乙醇之紅麴萃取物(3.08 mg/g , as gallic acid)顯著較高。綜合上述,經體外試驗結果顯示紅麴之乙醇萃取物皆具抗氧化特性,且呈現不同之抗氧化效力。添加不同發酵方式之紅麴發酵產物於低膽固醇飲食 (0.1%)中,可降低倉鼠血清之三酸甘油脂質、總膽固醇含量及低密度脂蛋白比率與增加肝臟/體重比。由肝臟組織病理切片觀察發現,添加適量紅麴發酵物可改善因膽固醇飲食造成之肝細胞脂肪變性現象(尤其是Mix 3組),而固態發酵組有色素輕微蓄積現象。補充適量紅麴發酵產物(0.03-0.18%)不影響倉鼠之肝指數(ALT, AST)及血尿素氮(BUN)含量。添加高劑量紅麴發酵物時(0.3%)則會降低動物存活率,且有膽管增生及中度近曲小管之肝細胞凝固性壞死等副作用(p0.05)。此研究新發現,添加適量紅麴發酵物,可顯著降低餵飼低膽固醇飲食之倉鼠血清中尿酸含量;此結果顯示,膳食補充紅麴可能影響尿酸代謝作用,值得進一步探討其影響機制。
針對紅麴發酵產物之延緩老化研究結果發現,飼料中添加紅麴發酵物(0.03%)可改善因老化引起老化促進小鼠(SAMP8)脊椎彎曲之症狀,亦可增加同月齡小鼠毛髮粗糙與柔軟度(p<0.05)。此外,補充紅麴發酵物可顯著降低雄鼠血液中總膽固醇、三酸甘油酯與尿酸含量,亦可降低雌鼠血液中低密度脂蛋白質之濃度(p<0.05)。無論雄雌鼠紅麴組之主動迴避試驗結果顯著均高於對照組,雄鼠腦部海馬迴區脂褐質堆積量顯著低於對照組,且肝臟總抗氧化力顯著高於對照組(p<0.05)。由以上結果證實,藉由飲食中添加紅麴發酵物可改善動物體內之氧化壓力,且有效地延緩SAMP8小鼠的老化,同時可以進一步改善11月齡SAMP8之學習記憶能力。
Monascus species have been confirmed to produce many metobiltes by cultivation on rice grains that is used as a functional dietary supplement. These metabolities includes hydrolytic enzymes, primary metabolites (unsaturated fatty acids, esters) and secondary metabolites (pigments, monascidin, monacolins, GABA, flavonoids and other unindentified bioactive compounds). Monascus-fermented product (MP) has been used as a functional dietary supplement and also as food material in various new products. Although several reports of improvement in functional characteristics by MP supplement have been issued, most of them focus on the prevention of hypertension and blood sugar. The purpose of this study is to investigate the antioxidative activity, thermal degradation kinetics of monacolin K, hypertriglyceridemic effects of Hamster rats, and learning and memory in SAMP8 mice.
Monacolin K content of MP solution remained fairly constant in the range of pH 3-9. The content of monacolin K rapidly decreased to 14.1% when heat temperature increased to 121℃ for 90min at pH 9. The thermal degradation kinetics of monacolin K followed a first order kinetic reaction (90-121℃, r2 = 0.99). The activation energy, enthalpy and entropy of monacolin K thermal degradation in 3% MP solution were estimated to be 126.64, 120.78 and 0.22 kJ/mol, respectively, as heated in the range of 90-121℃ (r2 = 0.99). Ethanolic extracts of MP has a dose-dependent kinetics in DPPH scavenging ability (DPPH IC50 = 6.67 ± 0.75mg/ml, r2 = 0.99). The highest reducing power was found to be 0.137 in 75% ethanol extracts of MP. The effects of MP and 0.1% cholesterol supplement on physiological characteristics suggested that MP significantly lowered total triglyceride, total cholesterol, and LDL ratio of serum total cholesterol but increased HDL, ratio of liver to BW in MP supplement diet groups (p<0.05). No side-effect on serum of ALT, AST, BUN level and morphology of tissues was observed in the MP 1 and MP 3 diet groups. As MP supplement level increased to 0.3% (high dosage diet), a significant side-effect was appeared on survival rate declining, bile ducts hyperplasia and moderate proximal tubular coagulative necrosis in liver of Hamster rats. This study indicated that all MP supplement diet groups significantly decreased the serum uric acid level (p < 0.05) as compared to control and normal diet groups. The results suggest the MP diets may promotion the purine metabolism. These findings conclude that MP supplement diet could improve the locomotion activity, grading score and active shuttle-avoidance test in both male and female SAMP8 mice (p < 0.05). It would appear to be a certain age-related changes as regards learning and memory can be improved by application of an MP-supplemented diet for SAMP8 mice. In the present study, these results have suggested that dietary supplementation of MP could be considered to effectively contribute to the retardation of the aging process for such individuals, as also contributing to an improvement in their learning ability, and encouraging a reduction in memory deterioration with age.
Contents
摘要 I
Abstract III
致謝 V
Contents VI
Table Contents IX
Figure Contents XI
Chapter 1.Literature Review 1
1. Monascus genus 2
1.1. Solid state cultivation 2
1.2. Submerged cultivation 2
2. Polyketides 4
3. Secondary metabolites of the fungus Monascus 4
3.1. Pigments 5
3.2. Antibiotic activity 6
3.3. Mevinolin and related compounds 7
3.4. γ–aminobutyric acid, GABA and acetylcholin 8
3.5. Flavonoids 9
4. Oxidative stress and healthy 10
4.1. Oxidative stress and its significance in biology 11
4.2. Redox imbalance and apoptosis 12
4.3. Oxidative stress in the brain and neurodegenerative disorders 13
Objectives of this study 15
Chapter 2.Antioxidative Capacity (in vitro) and Thermal Degradation Kinetics Analysis of Bioactivators in Monascus-fermented Products (MP) 30
Abstract 31
1. Introduction 32
2. Materials and methods 33
3. Results and discussion 37
3.1 Compositions of MP 37
3.2 DPPH free radical-scavenging ability 38
3.3. Reducing Power 39
3.4. Effect of pH and heat treatment on monacolin K 39
3.5. Thermal degradation of monacolin K 40
4. Conclusions 41
Chapter 3 Effect of Monascus-fermented Products Supplement on Physiological Characterization of Hamsters 54
Abstract 55
1. Introduction 56
2. Materials and methods 58
3. Results and discussion 60
3.1. Preliminary test 60
3.2. MP fermentation type effects on physiology of Hamsters (Exp.I ) 61
3.2.1. Biochemical analyses 62
3.2.2. MP effects on serum uric acid 64
3.2.3. Morphological analyses 65
3.3. Optimal dosage of MP supplementary (Exp. II) 66
Chapter 4 Supplementary of Monascus-fermented Products on Learning and Memory in the SAMP8 Mice 84
Abstract 85
1. Introduction 86
2. Materials and methods 88
3. Results 92
4. Discussion 94
Conclusion 107
References 108
Appendix Tables 123
Appendix Figures 126
Abbreviations 129
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