膳食果寡醣補充劑調節D-半乳糖注射Balb/c雄鼠體內腦部、心血管功能及抗氧化分子濃度

Dietary fructo-oligosaccharide supplementation modulated the neurological, cardiovascular function and antioxidant level in D-galactose-treated Balb/c mice

洪孟君(Meng-Jun Hong)

指導教授 ： 陳曉鈴

中山醫學大學/健康管理學院/營養學系碩士班/碩士(2011年)

摘要

目的: 本實驗主要研究目的在探討長期注射D-半乳糖是否誘發Balb/c雄性小鼠老化相關症候群，包括腦部β-amyloid形成與學習記憶功能，以及調節心血管功能和抗氧化分子狀態；以及飲食補充果寡醣或維生素E是否可以延緩這些老化現象。
方法: 將12週齡大Balb/c雄性小鼠分為控制組、D-半乳糖注射組、D-半乳糖低果寡醣組 (2.5% w/w)、D-半乳糖高果寡醣組 (5% w/w)及D-半乳糖維生素E組 (0.2% w/w)，飲食與皮下注射生理食鹽水 (控制組)或10 % D-半乳糖生理食鹽水 (1.2 g/kg BW)連續介入49天。另有自然老化組攝取控制組飼料，但是不接受注射，於64週齡大犧牲。實驗結束前5-7天以水迷宮測量老鼠之學習記憶功能，犧牲前3天測量心電圖及血壓。禁食12小時後於犧牲當天以CO2昏迷，開胸由右心房採血，並利用冰浴生理食鹽水進行組織灌流後收集腦部、心臟及肝臟，並測量左心室心肌厚度。血漿、肝臟儲存於-80℃以待後續分析血脂及抗氧化分子。腦部經固定及蔗糖溶液脫水後，進行冷凍切片及beta-amyloid的免疫染色。
結果：結果顯示，自然老化組及D-半乳糖組有相似的老化腦部功能退化情形，如學習記憶能力下降以及腦部皮質及海馬迴beta-amyloid堆積增加，補充果寡醣或維生素E可延緩腦部功能退化情形。於心血管方面，自然老化組及D-半乳糖組的心跳次數及左心室心肌厚度顯著高於控制組，而收縮壓、舒張壓及平均動脈壓有增加的趨勢。自然老化組及D-半乳糖組心電圖QRS區間及QTc有延長之情形，但是補充果寡醣或維生素E可使血壓及心電圖調節至類似控制組，且果寡醣在2.5% (w/w)時即可以看到效果，有劑量的效應。自然老化組及D-半乳糖組的血漿中總膽固醇及三酸甘油脂顯著高於控制組，而補充果寡醣或維生素E則可降低其濃度。於抗氧化物質方面，血漿維生素C濃度於各組間無顯著性的差異，但自然老化組及D-半乳糖組有較低趨勢，在給予補充果寡醣或維生素E時，可使其濃度提升。而血漿維生素E部分，控制組顯著性高於自然老化組及D-半乳糖組，而補充果寡醣則可提升其濃度。肝臟維生素E呈現跟血漿維生素E相同的結果。肝臟麩胺基硫(glutathione)總量，控制組有比自然老化組及D-半乳糖組高的趨勢，但無顯著性差異，而還原型GSH部分，控制組則顯著性高於自然老化組及D-半乳糖組，在給予補充果寡醣或維生素E時，皆可增加肝臟總GSH及還原型GSH含量。另外，GSSG/GSH 表示氧化壓力，發現自然老化組及D-半乳糖組顯著性高於控制組，補充果寡醣或維生素E則可以降低氧化壓力。
結論：本研究顯示D-半乳糖注射有效加速腦部功能與心血管功能衰退，以及血液及肝臟維生素E耗竭。補充果寡醣有助於及延緩D-半乳糖誘發之腦部及心血管功能病變，並維持血漿及肝臟抗氧化分子濃度的潛力。

關鍵字

D-半乳糖 ； 果寡醣 ； beta-amyloid ； 心血管功能 ； 抗氧化

並列摘要

Objectives: This experimental study long-term injection of D-galactose-induced Balb/ c mice associated with aging syndrome, including brain β-amyloid formation, learning and memory function, regulation of cardiovascular function and antioxidant status; and dietary supplement of fructo-oligosaccharide (FOS) or vitamin E can delay the aging process.

Materials and Methods：The twelve-week-old male Balb / c mice were divided into control group, D-galactose group, D-galactose low-FOS group (2.5% w/w), D-galactose high-FOS group ( 5% w/w) and D-galactose vitamin E group (0.2% w/w), diet and subcutaneous injection of saline (control group) or 10% D-galactose saline (1.2 g / kg BW) continuous involvement of 49 days. Another natural aging group of feed intake control diet, but not vaccinated, 64 weeks old great sacrifice. Experiment of end for 5-7 days to measure water maze (learning and memory function) and before 3 days measure ECG and blood pressure. Fasting bloods were collected from the heart and perfusion with ice-cold saline, after collection of brain, heart, liver, and measured left ventricular thickness. The plasma and liver were stored at -80 ℃ until subsequent analysis for blood lipids and antioxidant molecules. The brain tissue dehydrated of sucrose solution, the frozen sections and β-amyloid immunostaining.

Result: Natural aging group and D-galactose group have similar neurological degenerative of aging, such as learning and memory degeneration, brain beta-amyloid accumulation increased. Diet supplement FOS or vitamin E can delay the degenerative neurological conditions. In the cardiovascular function, natural aging group and D-galactose group the heart rate and left ventricular thickness was significantly higher than the control group, and systolic blood pressure, diastolic blood pressure and mean arterial pressure were increase. Natural aging group and D-galactose group the QRS and QTc intervals of ECG have extended, but added FOS or vitamin E can regulate blood pressure and heart function. Natural aging group and D-galactose group the plasma total cholesterol and triglycerides were significantly higher than the control group, supplement of FOS or vitamin E can decline blood lipids. In antioxidants, the plasma vitamin C concentration in each group no significant differences, but the natural aging group and D-galactose group have lower concentration, but supplement FOS or vitamin E can increase concentration. The plasma vitamin E of the control group was significantly higher than the natural aging group and D-galactose group whereas supplement of FOS or vitamin E can enhance the concentration. In the liver total glutathione (GSH), the control group than natural aging group and D-galactose group of high-trend, but no significant differences. The GSH level of the control group was significantly higher than those of the natural aging group and D-galactose group. FOS or vitamin E can increase liver total GSH and the reduced GSH content. The GSSG/GSH ratio of the natural aging group and D-galactose group was significantly higher than the control group, supplement FOS or vitamin E can reduce oxidative stress.

Conclusion: This study indicated that D-galactose injection accelerated decline of brain function and cardiovascular function, blood and liver vitamin E depletion. The dietary supplement of FOS can delay D-galactose-induced brain lesions and cardiovascular function, and maintain plasma and liver antioxidant of molecular concentration.

並列關鍵字

D-galactose ； fructo-oligosaccharide ； beta-amyloid ； cardiovascular function ； antioxidant

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