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研究生:吳惠青
研究生(外文):Hui- Ching Wu
論文名稱:益生孅-23複方產品對於餵食高膽固醇飲食倉鼠的脂質代謝及腸道菌相的影響
論文名稱(外文):Effects of PROBIO S-23 complex product on lipid metabolism and intestinal microflora of hamsters fed on high-cholesterol diet
指導教授:蔡政志蔡政志引用關係
指導教授(外文):Cheng-Chih Tsai
學位類別:碩士
校院名稱:弘光科技大學
系所名稱:食品暨應用生物科技所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
畢業學年度:99
語文別:中文
論文頁數:115
中文關鍵詞:益生菌倉鼠膽固醇高脂血症三酸甘油酯益生孅-23
外文關鍵詞:ProbioticsHamstersCholesterolHyperlipidemiaTriglyceridePROBIO S-23
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高脂血症是引起心血管疾病的危險因子之一,然而目前的藥物治療成本高且具有副作用等缺點,因此本研究擬開發降低膽固醇之益生菌輔助療法。主要目的為探討在高油脂高膽固醇飲食條件下,益生菌複方產品對於倉鼠之脂質代謝與腸道菌相的影響。將50隻雄性倉鼠分為五組,分別為控制組 (Control group),高油脂高膽固醇組 (High fat plus high cholesterol diet, HFC group) 以及低、中、高劑量益生孅-23 (PROBIO S-23) 之試驗組。控制組倉鼠餵食AIN-76基礎飼料,高油脂高膽固醇飼料配方為AIN-76基礎飼料,調配添加 12% 玉米油、3% 動物油及 0.5% 膽固醇。倉鼠經誘導十天後,使倉鼠成為高膽固醇血症狀態,開始進入實驗期,為期十週。在實驗期間,記錄體重及攝食量,每兩週收集一次血清樣本,分析膽固醇、三酸甘油酯、低密度脂蛋白、高密度脂蛋白和脂質過氧化指標硫代巴比妥酸反應物 (Thiobarbituric acid reactive substance, TBARS) 之濃度,並於第0和第9週收集新鮮糞便及犧牲後收集肝臟組織進行分析。結果顯示,PROBIO S-23低、中、高劑量組與HFC組之倉鼠相較,最終平均體重下降幅度分別為8.07%、11.36%、8.09%,在攝食方面,發現各組間倉鼠每日攝食量並無明顯差異。益生孅-23複方產品可顯著降低高膽固醇血症倉鼠的血清總膽固醇、低密度脂蛋白膽固醇、三酸甘油酯和TBARS濃度 (P<0.05)。但高密度脂蛋白膽固醇沒有顯著改變。而肝臟的總膽固醇、三酸甘油酯和TBARS的數值亦皆明顯下降 (P<0.05)。在肝臟功能檢測方面,血清肝指數麩草酸轉胺基酶 ( Glutamyl oxaloacetic transaminase, GOT)、麩丙酮酸轉胺基酶 (Glutamyl pyruvic transaminase, GPT) 與鹼性磷酸酶 (Alkaline phosphatase, ALP) 的數值亦明顯降低 (P<0.05),且白蛋白與球蛋白的比值 (Albumin/ Globulin ratio) 有升高的情況。此外,在第九週後,糞便中乳酸桿菌和雙歧桿菌的活菌數,餵食益生孅-23複方產品高劑量組與控制組做比較,分別增加2.77及1.4 對數值。綜合上述,證實益生孅-23複方產品具有改善高油脂高膽固醇飲食所引起的肥胖體質、脂質異常及肝臟機能損害的功效,亦可降低體內脂質過氧化物的生成,並增加腸道益生菌的數量,進而促進腸道微生物菌相的平衡。
Hyperlipidemia is considered as one of the risk factors to cause cardiovascular diseases. However, present medication appears the shortcomings of high cost and side effects. Therefore, this study tends to develop a complementary therapy with probiotics to reduce cholesterol. The aim of this study is to investigate the effects of probiotics complex products on lipid metabolism and intestinal microflora of hamsters with high fat plus high cholesterol diet. Fifty male hamsters were divided into five groups, including the control group, and the experiment groups with high fat plus high cholesterol diet (HFC group), low-, medium-, and high-dose PROBIO S-23. The control group were fed basal diet AIN-76 with 12% corn oil, 3% lard, and 0.5% cholesterol. After 10-day induction, these hamsters appeared hypercholesterolemia. The experiment then started for 10 weeks. The weights and the food intake were recorded. Serum was collected twice a week for analyzing the concentration of cholesterol, triglyceride, low density lipoprotein (LDL), high density lipoprotein (HDL), and the lipid peroxidation index such as thiobarbituric acid reactive substance (TBARS). Fresh faeces and liver were also collected on the zero and the ninth week for analysis. The results showed that the average weights of the groups with low-, medium-, and high-dose PROBIO S-23 reduced 8.07%, 11.36%, and 8.09%, respectively, in comparison with HFC group. In terms of daily food intake, there was no obvious difference among the hamsters in all groups. PROBIO S-23 complex products could remarkably decrease the concentration of total serum cholesterol, LDL cholesterol, triglyceride, and TBARS (P<0.05) of the hamsters with hypercholesterolemia. However, HDL cholesterol did not show significant difference. The total cholesterol, triglyceride, and TBARS in liver presented obvious decrease (P<0.05). In regard to the test on liver function, glutamyl oxaloacetic transaminase (GOT), glutamyl pyruvic transaminase (GPT), and alkaline phosphatase (ALP) appeared significant decrease (P<0.05); and the albumin/ globulin ratio was increased. Moreover, comparing the viable count of Lactobacillus and Bifidobacterium in the faeces after nine weeks, both the groups fed with high-dose PROBIO S-23 products and the control group increased 2.77 and 1.4 logarithms, respectively. In conclusion, PROBIO S-23 products improved the effect on obesity, abnormal lipid, and impaired liver function resulted from high fat plus high cholesterol diet. Besides, they could reduce the generation of lipid peroxide, increase the number of probiotics in intestine, and facilitate the balance of intestinal microbial.
目 錄

中文摘要 I
Abstract III
目 錄 V
表目錄 VIII
圖目錄 X
第壹章 前言 1
第貳章 文獻整理 3
一、血脂質及脂蛋白之介紹 3
(一)膽固醇 4
(二)三酸甘油酯 5
(三)乳糜微粒 6
(四)極低密度脂蛋白 6
(五)低密度脂蛋白 6
(六)高密度脂蛋白 7
二、高脂血症的定義及分類 7
(一)高脂血症之定義 7
(二)高脂血症之分類 8
三、心血管疾病 9
四、高脂血症與心血管疾病的相關性 13
五、心血管疾病與代謝症候群的相關性 13
六、代謝症候群與非酒精性脂肪肝的相關性 15
七、自由基對生物體之傷害 16
(一)自由基 16
(二)自由基來源 16
(三)活性氧 17
(四)脂質氧化傷害 17
(五)自由基與疾病關係 18
八、腸道菌相與健康 18
九、益生菌及益生質 21
(一)益生菌的定義 21
(二)益生質的定義 21
(三)益生菌和益生質的種類及功效 22
十、乳酸菌 23
(一)乳酸菌的定義 23
(二)乳酸菌的種類及應用 23
(三)乳酸菌之保健功效 24
(四)單株乳酸菌與多重乳酸菌株之比較 28
十一、 乳酸菌降膽固醇的功效 29
(一)降膽固醇的機制 29
(二)體外試驗 32
(三)動物試驗 33
(四)人體試驗 35
第参章 實驗架構 37
第肆章 材料與方法 38
一、實驗材料 38
(一)實驗動物 38
(二)飼料配製 38
(三)試驗樣品用量 40
(四)儀器設備 42
(五)選擇性培養基 42
二、實驗方法 44
(一)動物分組與飼養 44
(二)試驗動物致高血脂模式. 44
(三)試驗物質給予期間及途徑 44
(四)實驗動物樣品採集 45
(五)動物犧牲 45
(六)血液分析 46
(七)肝臟分析 52
(八)糞便菌相分析 53
(九)統計分析 53
第伍章 結果與討論 54
一、實驗期間倉鼠健康情況、體重、攝食量及臟器重量之變化 54
(一)外觀和活動力 54
(二)體重與攝食量變化 54
(三)臟器重量的變化 55
二、血液生化檢驗 55
(一)血脂質之變化 55
(二)血清脂質過氧化TBARS濃度的變化 59
(三)肝臟功能檢測 59
(四)腎臟功能檢測 61
(五)血中葡萄糖濃度之變化 62
三、肝臟檢測 62
(一)肝臟脂質之變化 62
(二)肝臟脂質過氧化TBARS濃度的變化 63
四、腸道糞便菌相之變化 64
第陸章 結論 65
第柒章 參考文獻 97


表目錄

表1. 人類血漿脂蛋白的特性與組成 3
表2. 血脂正常及異常值 8
表3. 中華民國血脂異常分類 9
表4. 代謝症候群之判定標準 14
表5. 實驗動物飼料組成份 39
表6. 實驗動物與人體表面積比等效劑量換算比率表 41
表7. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠體重增加及攝食量的影響 66
表8. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝重及相對肝重之影響 67
表9. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清總膽固醇的影響 68
表10. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清三酸甘油脂的影響 69
表11. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清高密度脂蛋白的影響 70
表12. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清低密度脂蛋白的影響 71
表13. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清脂質過氧化物含量的影響 72
表14. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清轉胺酵素活性的影響 73
表15. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清酵素活性的影響 74
表16. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清膽紅素的影響 75
表17. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清蛋白質的影響 76
表18. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清尿素氮、肌酸酐及尿酸濃度的影響 77
表19. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血中葡萄糖濃度的影響 78
表20. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝中膽固醇及三酸甘油酯含量的影響 79
表21. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝臟脂質過氧化物含量的影響 80
表22. 餵食不同濃度益生孅-23複方產品之倉鼠糞便中乳酸桿菌和雙岐桿菌之總菌數 81

圖目錄

圖1. 膽固醇之結構 4
圖2. 三酸甘油酯結構 5
圖3. 動脈粥狀硬化中血管內皮細胞功能失調 10
圖4. 動脈粥狀硬化中脂肪條紋的形成 11
圖5. 動脈粥狀硬化複雜性病變之形成 11
圖6. 動脈粥狀硬化中不穩定的纖維斑塊 12
圖7. 將保加利亞乳酸桿菌置於 (a) 不含膽固醇和(b)添加膽固醇之MRS中培養,利用掃描式電子顯微進行分析 30
圖8. 膽鹽水解酶降低膽固醇之機制 31
圖9. 餵食不同濃度益生孅-23複方產品和控制組十週期間倉鼠之體重變化 82
圖10. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清總膽固醇的影響 83
圖11. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清三酸甘油脂的影響 84
圖12. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清高密度脂蛋白的影響 85
圖13. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清低密度脂蛋白的影響 86
圖14. 餵食不同濃度益生孅-23複方產品十週期間對於高油脂高膽固醇飲食之倉鼠血清脂質過氧化物含量的影響 87
圖15. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清麩草酸轉胺基酶活性的影響 88
圖16. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清麩丙酮酸轉胺基酶活性的影響 89
圖17. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清鹼性磷酸酶活性的影響 90
圖18. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠血清白蛋白/球蛋白比值的影響 91
圖19. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝臟膽固醇含量的變化 92
圖20. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝臟三酸甘油脂含量的變化 93
圖21. 餵食不同濃度益生孅-23複方產品十週後對於高油脂高膽固醇飲食之倉鼠肝臟脂質過氧化物含量的影響 94
圖22. 餵食不同濃度益生孅-23複方產品之倉鼠糞便中Lactobacillus之菌數 95
圖23. 餵食不同濃度益生孅-23複方產品之倉鼠糞便中Bifidobacterium之菌數 96


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