臺灣博碩士論文加值系統

根據衛生福利部公布 106 年十大死因中糖尿病為第 5 名(衛生福利部，2017)，因此尋求治療糖尿病的天然藥物成為研究焦點。目前在糖尿病患者中約 90%罹患二型糖尿病，若只抑制血糖而沒有恢復胰島素接受體敏感性是無法治本的。因此本研究將探討以牛蒡、葛根、藿香、桑葉、玉竹、覆盆子及甜菊葉組成的牛蒡複方萃取物( BUC )對二型糖尿病的作用。長期胰島素阻抗會導致胰臟細胞的衰竭，因此本實驗先以大鼠的胰臟細胞( RIN-m5f )進行細胞增生率試驗，探討 BUC 對 β 細胞之增生效果，並進一步以動物實驗探討 BUC 調節血糖的作用機轉。由實驗結果得知，在給予 25.0 μg/mL的 BUC 作用 24 小時後，RIN-m5f 的增生率可達: 124.40 ± 0.18 % 。在動物實驗中，連續投予 28 天兩種不同劑量( 50.0、 100.0 mg/kg )的 BUC 後，發現 STZ 誘發組及 50.0 mg/kg 在生化指標上(總蛋白、血糖、三酸甘油脂、總膽固醇)有明顯的差異性。另外，本實驗以胰島素、胰島素接受體、昇糖素類似胜肽-1三種蛋白探討降血糖機制，結果發現胰島素接受體在 BUC 低劑量組的作用下有明顯增加，且含量高於誘發組，含量分別為: 0.76 ± 0.03； 0.95 ± 0.03 ng/mL。為深入探討胰島素接受體含量上升之原因，另以小鼠進行後續作用路徑的研究，根據實驗結果得知: 在給予連續 28 天的 BUC 作用下，STZ 誘發組及 BUC 100.0 mg/kg 組在小鼠的總蛋白、血糖、三酸甘油脂及總膽固醇等指標，出現統計差異。為探討胰島素接受體活性增加的原因，以 Insulin receptor substrate 2、 Serine/threonine protein kinase 1/2/3、 Glucose transporter type 4 三種蛋白進行 BUC 調節血糖作用機轉的分析，結果發現三種蛋白在 BUC 100.0 mg/kg 作用下，含量比 STZ 誘發組高，與誘發組相比分別增加 70.0、 66.9、 72.7%。綜合上述結果得知: BUC 具有調節血糖的作用，尤其在 100.0 mg/kg 作用下呈現最好的效果。BUC 的作用機轉是藉由增加 IRS2 活性而調控 β 細胞的增生；且透過增加胰島素接受體含量及下游蛋白 AKT1/2/3、 GLUT4 的活性，促進葡萄糖的代謝。故 BUC 確實具有開發為預防二型糖尿病保健產品的潛力。

According to statistics from the Ministry of Health and Welfare, diabetes ranks fifth among the top ten death diseases in 106 years. The search of natural medicines for treating diabetes has become an urgent issue. It is currently known that about 90% of all diabetic patients are type 2 diabetes. If only suppressing blood glucose and not restoring insulin receptor sensitivity, it is impossible to cure the disease! Therefore, this study explored how he burdock compound (BUC) composed of burdock, puerariae radix, cablin patchouli herb, mulberry leaf, polygonati odorati rhizoma, raspberry and stevia rebaudiana leaves to improve type　2　diabetes. Long-term insulin resistance lead to the destruction of pancreatic cells and thus affect the secretion of insulin. In this experiment first take MTT test on rat pancreatic cells (RIN-m5f) to investigate the effect of BUC on the proliferation of β cells, and experiment with animals further. The study in-depth discuss on the effect of BUC on regulation of blood glucose. According to the experimental results, the proliferation rate of RIN-m5f can reach 124.40 ± 0.18 % after 24 hours of administration of BUC 25.0 μg/mL. In the animal experiment, after 28 days of continuous administration of BUC at two different doses (50.0, 100.0 mg/kg), the induced groups and 50.0 mg/kg were found to have statistically significant differences in biochemical parameters (total protein, blood glucose, triglyceride, total cholesterol). In addition, insulin, insulin receptor and glucagon-like peptide-1 proteins were used to investigate the mechanism of hypoglycemic activity. It was found that insulin receptors were significantly increased in the 50.0 mg/kg of BUC. Compared with the induction group, the insulin receptor contents were 0.76 ± 0.03; 0.95 ± 0.03 ng/mL, respectively. In order to further search the reasons for the increase in insulin receptor content, we took the follow-up pathway in mice. According to the experimental results, the induction group and the BUC 100.0 mg/kg group in the 28 days BUC treatment, there were also statistical differences in the biochemical indicators of the mice (total protein, blood glucose, triglycerides, total cholesterol). In order to investigate the cause of the increase in insulin receptor activity, the three proteins of insulin receptor substrate 2, serine/threonine protein kinase 1/2/3 and glucose transporter type 4 were used to analyze the effect of BUC on blood glucose regulation. Three proteins were found in BUC 100.0 mg/kg the content was higher than that of the induced groups, which increased respectively by 70.0, 66.9, and 72.7%, compared with the induced groups. Based on the above results, especially at the dose of 100.0 mg/kg, BUC exhibited the effect of regulating blood glucose. The mechanism of action of BUC regulates the proliferation of β cells by increasing the activity of IRS2. BUC through recover the insulin receptor content and activity of downstream protein AKT1/2/3、 GLUT4, that promoted glucose metabolism. BUC have the potential to develop health product for the prevention of type 2 diabetes.

誌謝 I
目 錄 II
表次目錄 IV
圖次目錄 V
摘要 VII
縮寫表 X
第壹章　緒言 1
第貳章　文獻回顧 2
第一節 糖尿病 2
第二節 牛蒡複方萃取物簡介及藥理活性 15
第三節 鏈脲佐菌素(Streptozotocin, STZ)致病機轉 35
第四節 胰島素接受體 37
第五節 胰島素阻抗成因 41
第六節 研究動機與目的 44
第參章　材料與方法 46
第一節 實驗儀器及藥品 46
第二節 材料製備 51
第三節 細胞培養 52
第四節 動物試驗 55
第肆章　結果 67
第一節 牛蒡單方萃取物對大鼠胰臟細胞 RIN-m5f 增生率試驗之影 響 67
第二節 牛蒡複方萃取物對大鼠胰臟細胞 RIN-m5f 增生率試驗之影 響 69
第三節 比較牛蒡單方及複方萃取物對大鼠胰臟細胞 RIN-m5f 增生 率試驗之影響 71
第四節 牛蒡單方及複方萃取物對 STZ 誘導糖尿病小鼠血糖之影響 72
第五節 牛蒡複方萃取物對 STZ 誘導糖尿病大鼠之影響及機轉探討 73
第六節 牛蒡複方萃取物對 STZ 誘導糖尿病小鼠之影響及機轉探討 91
第伍章　討論 106
第陸章　結論 108
參考文獻 110

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