蝦殼富含蛋白質、幾丁質和蝦紅素。本實驗室由南美白蝦 (Litopenaeus vannamei) 篩選而得Lactobacillus plantarum LV3204、Stenotrophomonas maltophilia LV2122 (蛋白質分解菌) 及Aeromonas dhakensis LV1111 (幾丁質分解菌)，以此三株複合菌株液態發酵蝦殼。本研究旨探討蝦殼發酵產物的活性物含量，並探討餵食發酵蝦殼對第二型糖尿病大鼠之調節血糖與血脂功效。凍乾發酵蝦殼之總胜肽與蝦紅素含量分別為 16.36 與 1.13 mg/g，為未發酵蝦殼之 13.79 與 2.83 倍。雄性 Sprague - Dawley (SD) 大鼠除了正常控制組 (NC) 外，其餘以nicotinamide (NA) (230 mg/kg B.W.) - streptozotocin (STZ) (65 mg/kg B.W.) 誘導為第二型糖尿病大鼠，將其分為糖尿病控制組 (DC)、餵食未發酵蝦殼粉組 ( 9,000 mg/k g BW ,USS9000)、餵食低劑量發酵蝦殼粉組 (3000 mg/kg BW, FSS3000) 及高劑量發酵蝦殼粉組 (9,000 mg/kg BW, FSS9000) 及餵食蝦紅素標準品組 (20 mg/kg BW ，AST20)。餵食 7 週後，FSS9000 組之 OGTT 120 min血糖、空腹血糖及 HOMA-IR (Homeostasis Model Assessment-Insulin Resistance index) 均分別比 DC 組降低27.59%、22.54% 及50%。此外，FSS9000 組的相對脂肪重量 (Relative adipose weights)、血漿及肝臟 Total cholesterol (TC)、Triglyceride (TG)、及過氧化值均顯著降低。FSS9000 組肝臟六碳糖激酶 (Hexokinase) 與葡萄糖-6-磷酸脫氫酶 (Glucose-6-phosphate dehydrogenase) 分別比 DC 組提升 75.03% 與 83.16%，且其葡萄糖-6-磷酸酶活性顯著降低。此外，餵食發酵蝦殼顯著降低大鼠肝臟 Acetyl-CoA carboxylase 與 Fatty acid synthase 活性，並顯著提升脂肪組織 Hormone sensitive lipase 及 Lipoprotein lipase 活性。綜上所述，以複合菌株發酵蝦殼產品具作為調節血糖、血脂保健食品之潛力。

Shrimp shells (SS) are rich in protein, chitin and astaxanthin. Lactobacillus plantarum LV3204, Stenotrophomonas maltophilia LV2122 (proteolytic bacteria) and Aeromonas dhakensis LV1111 (chitin-decomposing bacteria) were screened from the white shrimp (Litopenaeus vannamei) in our laboratory, and these three composite strains were used to submerged ferment SS. The aim of this study were to investigate the active substance content of the fermented SS, and to conduct a feeding test on type 2 diabetic rats to explore the effect of fermented SS on hypoglycemic and hypolipidemic activity. The total peptide and astaxanthin contents of the fermented SS were 16.36 and 1.13 mg/g, respectively, which were 13.79 and 2.83 times that of the unfermented group. This shrimp shells fermented product was lyophilized into powders, and fed to type 2 diabetic rats to explore its hypoglycemic activity and hypolipidemic. The contents of total peptides and astaxanthin in freeze-dried fermented shrimp shells were 16.36 and 1.13 mg/g, respectively, which were 13.79 and 2.83 times that of unfermented shrimp shells. Except the normal control group (NC), male Sprague-Dawley (SD) rats were treated with nicotinamide (NA) (230 mg/kg B.W.) - streptozotocin (STZ) (65 mg/kg B.W.) to induce type 2 diabetes mellitus. These induced rats were divided into diabetes control group (DC), unfermented SS group (9,000 mg/kg BW, USS9000), low-dose (3000 mg/kg BW, FSS3000) and high-dose (9000 mg/kg BW, FSS9000) fermented SS groups, and astaxanthin standard group (20 mg/kg BW, AST20). After 7 weeks of feeding experiment, the values of OGTT 120 min blood glucose, fasting blood glucose and HOMA-IR (Homeostasis Model Assessment-Insulin Resistance index) in the FSS9000 group were significantly reduced by 27.59%, 22.54%, 50%, respectively, compared to those in the DC group. In addition, relative adipose weights, plasma and liver total cholesterol (TC), triglyceride (TG), and peroxide values were significantly reduced in the FSS9000 group. Compared with the DC group, the levels of hepatic Hexokinase and Glucose-6-phosphate dehydrogenase in the FSS9000 group were increased by 75.03% and 83.16%, respectively, and the activity of Glucose-6-phosphate was significantly decreased. In addition, feeding fermented SS significantly reduced the activities of Acetyl-CoA carboxylase and Fatty acid synthase in rat liver, and significantly increased the activities of hormone sensitive lipase and lipoprotein lipase in adipose tissue.
In conclusion, the fermented shrimp shell products with compound strains have the potential for a health food with hypoglycemic and hypolipidemic activity.

摘要 II
Abstract III
表目錄 xi
圖目錄 xii
附錄 xiv
壹、前言 1
貳、文獻整理 3
一、 糖尿病 3
1. 糖尿病之定義與分類 3
1.1 第一型糖尿病 (Type 1 diabetes mellitus) 3
1.2 第二型糖尿病 (Type 2 diabetes mellitus) 3
1.3 其他特異型糖尿病 (Other specific types of diabetes) 4
1.4 妊娠糖尿病 (Gestational diabetes mellitus, GDM) 4
2. 糖尿病診斷標準 5
3. 糖尿病對醣類代謝之影響 5
4 糖尿病併發症與治療 6
5. 第二型糖尿病動物誘導模式 6
5.1 Streptozotocin 誘導高血糖機制 7
5.2 Nicotinamid胰臟 β 細胞保護機制 8
5.3 Streptozotocin - Nicotinamide誘導第二型糖尿病機制 8
二、 胰島素 8
1. 胰島素訊息傳遞及其作用機制 9
2 胰島素阻抗 (Insulin resistance, IR) 9
三、 高脂/膽固醇飲食 10
1. 高脂飲食的定義 10
2. 高脂飲食對脂質代謝之影響 10
2.1 膽固醇的基本介紹 10
2.2 血脂質代謝 11
2.3 肝臟脂質代謝 12
2.4 下視丘 (Hypothalamic) 調控脂肪酸代謝 12
2.5 小腸維持脂肪代謝平衡 13
3. 脂肪組織脂質代謝相關酵素 13
3.1 荷爾蒙敏感性脂解酶 (Hormone-sensitive lipase, HSL) 13
3.2 脂蛋白脂解酶 (lipoprotein lipase, LPL) 14
四、 蝦殼 15
1. 蝦殼成分組成 15
1.1 幾丁質 17
1.2 蛋白質 18
1.3 類胡蘿蔔素 19
1.4 礦物質 19
2. 蝦殼之降血糖活性成分 19
2.1 蝦紅素 19
2.2 幾丁質衍生物 20
2.3 活性胜肽 21
2.4 其它 21
3. 蝦殼發酵 22
3.1 菌株篩選與發酵條件探討 22
參、實驗架構 24
一、 發酵蝦殼製備 24
二、 動物實驗 25
三、 犧牲後分析項目 26
肆、實驗材料與方法 27
一、實驗材料 27
1. 材料 27
2. 實驗菌株 27
3. 化學藥品 27
4. 培養基 28
5.動物實驗 28
5.1. 實驗動物 28
5.2. 實驗飼料 28
5.3. 分析套組 29
二、儀器設備 30
三、實驗方法 32
1. 菌株保存與活化 32
1.1 菌株保存 32
1.2 菌株活化 32
1.3 菌種鑑定 32
2. 實驗樣品製備與分析 32
2.1 蝦殼前處理 32
2.2蝦殼發酵基質配製 33
2.3 蝦殼發酵條件 33
2.4 分析 33
2.4.1 菌數分析 33
2.4.2 pH值 33
2.4.3 可滴定酸度分析 33
2.4.4 游離胺基態氮 34
2.4.6總胜肽含量 34
2.4.7蝦紅素含量 35
2.4.8 幾丁質含量 35
2.4.9 幾丁寡醣含量 36
2.5 一般成分組成 36
2.5.1 水分 37
2.5.2 灰分 37
2.5.3 粗脂肪 37
2.5.4 粗蛋白 37
2.5.5 碳水化合物 38
3. 動物飼養與分組 38
4. 動物犧牲及取樣 39
5. 動物實驗分析 40
5.1 血漿分析 40
5.1.1空腹血糖及葡萄糖耐量試驗 40
5.1.2血漿中總膽固醇濃度測定 40
5.1.3血漿中三酸甘油酯濃度測定 41
5.1.4血漿中胰島素含量測定 41
5.1.5血漿中天門冬氨酸胺基轉胺酶 (Aspartate aminotransferase, AST) 、丙氨酸胺基轉胺酶 (Alanine aminotransferase, ALT) 活性測定 42
5.1.6血漿中肌酸酐 (Creatinine) 含量測定 43
5.1.7血漿中尿素氮 (Blood urea nitrogen, BUN) 含量測定 43
5.1.8血漿中脂蛋白 (Lipoprotein) 分離與測定 44
5.2 肝臟分析 44
5.2.1 肝臟脂質萃取 44
5.2.2 肝臟總膽固醇及三酸甘油酯測定 45
5.2.3 肝臟 TBARS 含量測定 45
5.2.4 肝臟酵素活性測定-肝臟細胞質液製備 45
5.2.5 肝臟細胞質液蛋白濃度測定 46
5.2.7 六碳糖激酶 (Hexokinase, HK) 活性測定 46
5.2.8 葡萄糖 - 6 - 磷酸酶 (Glucose - 6 - phosphatase, G6Pase) 活性測定 47
5.2.9 葡萄糖 - 6 - 磷酸去氫酶 (Glucose - 6 - phosphate dehydrogenase, G6PD) 活性測定 48
5.2.10 脂肪酸合成酶 (fatty acid synthase, FAS) 活性測定 49
5.2.11 乙醯輔酶 A 羧化酶 (acetyl-CoA carboxylase, ACC) 活性測定 50
5.3 脂肪分析 51
5.3.1 脂肪細胞質液製備 51
5.3.2 荷爾蒙敏感性脂解酶 (Hormone sensitive lipase, HSL) 含量測定 51
5.3.3 脂蛋白脂解酶 (lipoprotein lipase, LPL) 含量測定 53
5.4 小腸分析 53
5.4.1 收集小腸黏膜 53
5.5 糞便分析 54
5.5.1 糞便乾重及濕重 54
5.5.2 糞便中脂質萃取 55
5.5.3 糞便中總膽固醇與三酸甘油酯分析 55
5.6 攝食量、飲水量及排尿量 55
6. 統計分析 55
伍、結果與討論 56
陸、結論 69
柒、參考文獻 71
捌、圖表 91
玖、附錄 116

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