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研究生(外文):Nai-Qi Chen
論文名稱(外文):Studies of molecular mechanisms of NQ-1 on the hypoglycemic effect in vitro and in vivo
外文關鍵詞:PPAR dual agonisttypeⅡdiabetes mellitusinsulin resistance
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過氧化物酶體增殖物激活受體(peroxisome proliferator-activatived receptors,簡稱PPARs)是一群需要配體(ligand)激活的轉錄因子,屬於核激素受體超家族,在許多生理作用中扮演重要的角色,例如:提高胰島素的敏感性、調節脂質代謝、維持體內葡萄糖平衡、細胞週期的調控與發炎反應的調節。在過去幾年中,對於天然和合成的過氧化物酶體增殖物激活受體配體的開發,及其相關的功能之鑑別,已經成為糖尿病病人血糖控制重要的研究目標之一。NQ-1是從中草藥純化出來的一種疏水性化學物質。首先,我們從細胞轉染(transition transfection assay)實驗中發現隨著NQ-1劑量的提高細胞中PPARa及g的活性亦顯著地增加。接著,以西方墨點法觀察,NQ-1可以分別在3T3-L1脂肪細胞及人類肝癌細胞(HepG2)中誘導PPARa/g產生。除此之外,NQ-1也可顯著地誘導3T3-L1脂肪細胞之脂肪標的基因表現(adipocyte marker gene),例如:脂肪酸結合蛋白(adipose fatty acid-binding protein, aP2)、脂蛋白脂解酶(lipoprotein lipase, LPL)、脂肪酸合成酶(fatty acid synthase, FAS)和CCAAT/enchancer binding protein-a(C/EBPa)。第三,NQ-1同樣地誘導HepG2肝癌細胞表現PPARa標的基因增加,例如: CPT-1與ACS。最後,在動物實驗方面,糖尿病小鼠(db/db mice)以腹腔注射的方式投予NQ-1,發現NQ-1能夠降低血糖卻無法降低血漿中胰島素與三酸甘油脂的濃度。由上述的實驗結果顯示,NQ-1為PPARa/g的雙重配體,並且具有治療第二型糖尿病的可能性。
Peroxisome proliferator-activatived receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-dependent transcription factors. PPARs play a central role in insulin sensitivity, lipid metabolism, glucose homeostasis, cell cycle progression, and inflammation. For the control blood glucose level of diabetes mellitus patient, searching PPAR activators from natural and synthetic chemicals, and discovering their associated functions is one of the most important works in the field. NQ-1 is a hydrophobic chemical and extracted from Chinese herb medicine. First, NQ-1 significantly activated both of PPARa and PPARg in a dose-dependent manner by transition transfection assay. Second, NQ-1 markedly induced the expression of PPARa and PPARg in HepG2 cells and 3T3-L1 respectively. In addition, NQ-1 significantly induced the expression of adipogenic factors such as aP2, LPL, C/EBPa and fatty acid synthase in 3T3-L1cells. Third, NQ-1 also induced the expression of PPARa target genes such as CPT-1 and ACS in HepG2 cells. Finally, obese diabetic mice (db/db mice) were treated with NQ-1, and found that NQ-1 markedly reduced plasma glucose level, however did not change the plasma insulin level and serum triglyceride level. These results suggest that NQ-1 is a dual PPARa/g activator and may have a beneficial effect on the therapy of typeⅡdiabetes mellitus.
中文摘要 Ⅴ
英文摘要 Ⅵ
第一章  緒論 1
1-1 前言 1
1-2 認識糖尿病 2
1-2-1 糖尿病的分類與檢測 2
(1) 糖尿病的診斷標準 2
(2) 糖尿病的分類 2
1-2-2糖尿病治療方針 4
1-2-3 胰島素抗性 6
1-3 過氧化物酶體增殖物激活受體 (PPARs) 9
1-3-1 PPARs簡介 9
1-3-2 PPARa的簡介 10
(1) PPARa與脂質代謝 11
(2) PPARa與動脈粥樣硬化 11
(3) PPARa與胰島素抗性 11
(4) PPARa與發炎反應 12
1-3-3 PPARg的簡介 12
(1) PPARg的配體 13
(2) PPARg對抗發炎反應的影響 15
(3) PPARg對癌症的影響 15
(4) PPARg對細胞週期的影響 16
1-3-4 PPARb的簡介 16
1-4 實驗動物介紹 17
研究目標 18
第二章 實驗材料與方法 19
2-1 實驗試劑 19
2-2 儀器 20
2-3 抗體 21
2-3-1 一級抗體 21
2-3-2 二級抗體 22
2-3-3 引子序列 22
2-4 實驗方法 24
2-4-1細胞培養與分析 24
(1) 解凍細胞 24
(2) 細胞繼代培養 24
(3) 脂肪細胞分化培養 25
(4) Oil red O染色及脂肪定量 25
(5) 轉染試驗 25
2-4-2西方墨點法 26
2-4-3反轉錄聚合酶酵素連鎖反應 28
(1) 純化RNA 28
(2) 反轉錄 28
(3) 聚合酶酵素連鎖反應 28
2-4-4 動物實驗 29
(1) 實驗動物 29
(2) 實驗方法 29
(3) 免疫化學組織染色切片 30
(4) 脂肪組織切片及染色 31
2-4-5 統計方法 31
第三章 結果 32
3-1 NQ-1可以活化PPARa與PPPARg 32
3-2 NQ-1誘發脂質新生 32
3-3 NQ-1誘導PPARa及其標的基因的表現 33
3-4 NQ-1誘導PPARg及其標的基因的表現 33
3-5 NQ-1降低db/db mice的血糖值 34
第四章 討論 35
第五章 參考文獻 38
Figure 1. Effect of NQ-1 on the activation of PPARa and PPARg in cells 45
Figure 2. Effects of NQ-1 on adipocyte differentiation 46
Figure 3. Effects of NQ-1 on the expressions of PPAR itself and its target genes in cells 47
Figure 4. Effects of NQ-1 on plasma glucose, insulin and TG levels in db/db mice 50
Figure 5. Effects of NQ-1 on the pancreatic insulin content and morphology of white adipose tissue
Table 1. Effects of NQ-1 on the body weight and food intake in db/db mice 49
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