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研究生: 陳俐彣
Chen, Li-Wen
論文名稱: 臺灣天仙果萃取物減緩去卵巢糖尿病小鼠周邊神經病變之研究
Alleviative Effect of Ficus formosana Extract on Peripheral Neuropathy in Ovariectomized Diabetic Mice
指導教授: 沈賜川
Shen, Szu-Chuan
吳瑞碧
Wu, Swi-Bea
丁俞文
Ting, Yu-Wen
口試委員: 沈賜川
Shen, Szu-Chuan
吳瑞碧 丁俞文 吳忠信 黃文忠 趙伯寬
口試日期: 2021/08/16
學位類別: 碩士
Master
系所名稱: 營養科學碩士學位學程
Graduate Program of Nutrition Science
論文出版年: 2021
畢業學年度: 109
語文別: 中文
論文頁數: 82
中文關鍵詞: 神經病變臺灣天仙果去卵巢糖尿病鼠抗發炎
英文關鍵詞: Neuropathy, Ficus formosana Maxim, ovariectomized, diabetic mice, anti-inflammatory
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202101511
論文種類: 學術論文
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  • 國人糖尿病盛行率逐年攀升,而更年期婦女因女性賀爾蒙分泌紊亂易造成血糖的波動,為罹患糖尿病重要的高風險族群。神經病變為臨床常見的糖尿病合併症之一,糖尿病患者易因氧化壓力的增加導致末梢周邊神經受損等病變,起初會有蟲爬、疼痛等感覺過敏之情形,當神經完全凋亡時即會失去知覺並造成行動上的障礙。臺灣天仙果 (Ficus formosana Maxim.) 為桑科榕屬植物,生長於臺灣中低海拔山坡地,為坊間常見之食補藥材,具強筋健骨、調整經期等功效。本研究以去卵巢糖尿病雌性小鼠模擬更年期糖尿病婦女,探討臺灣天仙果熱水萃取物減緩其神經病變之效果。透過給予C57BL/6J去卵巢雌性小鼠餵食60%高脂飲食及施打低劑量STZ誘導為糖尿病,並以自由採食或管餵方式(含萃取物200、2000ppm)分別連續處理6週後,觀察小鼠血糖、神經相關行為之變化、足底末梢及坐骨神經病理切片以及發炎疼痛相關mRNA表現。結果顯示,自由採食高劑量(2000ppm)臺灣天仙果萃取物飼料可以顯著改善去卵巢糖尿病小鼠負控制組之血糖(p<0.05);去卵巢糖尿病小鼠不論是以飼料或是管餵給予臺灣天仙果萃取物處理均能有效改善小鼠腳掌過度敏感、減緩C纖維的異常增生和斷裂、坐骨神經損失、減少氧化損傷之情形發生(p<0.05),表示臺灣天仙果莖部萃取物可延緩去卵巢糖尿病小鼠腳掌因末梢神經受損所導致之過度敏感反應。文獻指出,坐骨神經中發炎相關激素的大量表達會引發疼痛感覺,本研究發現不論以自由採食飼料或是管餵(200、2000 ppm)天仙果萃取物處理,均能顯著降低去卵巢糖尿病小鼠坐骨神經中IL-1β、IL-6、IFN-γ、COX-2等發炎相關激素mRNA表現量 (p<0.05)。推測這可能是減緩疼痛及敏感的關鍵機制。本研究之結果可作為臺灣天仙果開發糖尿病神經病變藥物時參考,而其主要活性化合物及發炎與疼痛詳細機制有待未來進一步探討。

    The prevalence of diabetes is increasing year by year in Taiwan. The menopausal women are prone to have fluctuations in blood sugar due to unstable secretion of female hormones, making them to be one of the high risk groups on suffering diabetes. Neuropathy is a common clinical diabetic complication. The peripheral nerves will be gradually damaged due to the increase of oxidative pressure. Initially, patients will be hyperesthesia, such as insect crawling, pain, etc., and lost feeling and action abilities when the nerves are completely apoptosis. Ficus formosana Maxim is a ficus plant in the moraceae family. It mainly grows on the slopes of low and middle-altitude hills in Taiwan. In this study, ovariectomized mice with type 2 diabetes were used to mimic menopausal diabetic women, and the effect of Ficus formosana Maxim extract (FFE) on reducing neuropathy in ovariectomized diabetic mice was invaluated. C57BL/6J ovariectomized female mice were fed high-fat diet (60% calorie of diet) and injected low-dose STZ (50mg/kg BD) to induce type 2 diabetes, and were given the extract daily by ad libitum and tube feeding FEE at dosages of 200, 2000ppm, respectively, for 6 weeks. At the end of animal trial, the blood chemicals, nerve-related behaviors, pathological sections of the intraepidermal and sciatic nerves, inflammation and pain response-related mRNA expressions in mice were analyzed. The results showed that the blood glucose level of high-dose FFE group significantly decreased compared to the STZ group (p<0.05). Moreover, FFE significantly alleviated the hypersensitivity, abnormal proliferation and rupture of C fiber, loss of sciatic nerve, and reduce oxidative damage in ovariectomized diabetic female mice. The high-dose FFE administered by tube shows the higest benefit (p<0.05) and is better than lipoic acid, which is currently used for diabetic neuropathy in the European Union. In this study, the administration of FFE significantly decreased the mRNA expression of IL-1β, IL-6, IFN-γ, COX-2 in diabetic mice (p<0.05). We speculated it may be associated to the key mechanism on alleviating pain and sensitivity, and the active compound may be associated to the abundant polyphenols of FFE. The further investigation on detail mechanisms of inflammation and pain, as well as the active compounds of FFE are necessary to evaluate the potential of Ficus formosana Maxim as a therapeutic drug of diabetic peripheral neuropathy.

    第一章 前言 1 第二章 文獻回顧 第一節. 糖尿病 一、糖尿病定義、流行病學 2 二、胰島素與胰島素阻抗 3 三、糖尿病病徵、分類 4 四、糖尿病臨床診斷 6 第二節 糖尿病周邊神經病變 一、糖尿病周邊神經病變之診斷、流行病學 7 二、周邊神經結構與功能 9 三、氧化壓力對周邊神經之影響 10 第三節 發炎與疼痛性糖尿病神經病變 一、疼痛傳遞機制 13 二、神經受損與發炎疼痛反應 14 三、電壓門控鈉離子通道 14 四、臨床上疼痛性糖尿病神經病變藥物 16 第四節 糖尿病與更年期婦女神經病變 一、更年期定義、症狀 17 二、更年期婦女賀爾蒙之變化 17 三、更年期婦女與糖尿病神經病變之關聯 19 第五節 臺灣天仙果(Ficus formosana Maxim) 一、臺灣天仙果簡介 20 二、臺灣天仙果成分 20 三、臺灣天仙果之藥理作用 20 四、臺灣天仙果莖部熱水萃取物治療糖尿病周邊神經病變之潛力 22 第三章 研究動機及實驗架構 第一節 研究動機與目的 23 第二節 實驗架構 24 第四章 實驗材料與方法 第一節 實驗藥品與儀器 一、 實驗藥品與試劑 25 二、 儀器設備 25 第二節 樣品來源與成分 一、 實驗樣品來源與製備 26 二、 實驗樣品成分 27 第三節. 動物試驗設計 一、實驗動物 27 二、實驗動物飼料 27 三、實驗動物誘導與分組 28 四、實驗方法 29 五、統計分析 34 第五章 結果 第一節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠血糖之影響 一、臺灣天仙果莖部熱水萃取物介入第零週之血糖值 35 二、臺灣天仙果莖部熱水萃取物介入六週後之血糖值 35 三、臺灣天仙果莖部熱水萃取物介入六週後血糖變化值 35 第二節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠行為之影響 一、腳掌機械刺激測試-Von Frey Filament testing 39 二、溫度感應測試Thermal perception testing 39 第三節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠病理切片之影響 一、足部表皮神經分佈 42 二、坐骨神經病理切片 42 第四節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠胰島素阻抗、髓鞘合成相關蛋白mRNA表現之影響 一、胰島素受體底物-2(IRS-2)mRNA表現量 49 二、生長因子受體蛋白-2(Grb2)mRNA表現量 49 第五節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠發炎相關蛋白mRNA表現之影響 一、腫瘤壞死因子-α(TNF-α)mRNA表現量 52 二、干擾素-γ (IFN-γ) mRNA表現量 52 三、白細胞介素-6(IL-6)mRNA表現量 55 四、白細胞介素-1β(IL-β)mRNA表現量 55 五、環氧合酶-2(Cox-2)mRNA表現量 55 第六節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠發炎疼痛鈉離子通道相關蛋白mRNA表現之影響 一、電壓門控鈉離子通道-Nav1.8 mRNA表現量 59 二、蛋白激酶-ε (PKC- ε) mRNA表現量 59 第六章 討論 第一節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠血糖與血液生化代謝指標之影響 一、胰島素、血糖變化 62 二、血脂變化 62 第二節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠行為試驗及足底末梢神經之影響 一、行為試驗之影響 63 二、足底末梢神經之變化 64 三、保護足底末梢的可能機制 64 第三節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠坐骨神經氧化傷害及髓鞘合成之影響 一、減緩氧化傷害 65 二、髓鞘合成相關蛋白之mRNA表現 65 第四節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠之抗發炎效應 66 第五節 臺灣天仙果莖部熱水萃取物對去卵巢糖尿病雌性小鼠之疼痛通道mRNA表現之影響 67 第七章 結論 69 參考文獻 71 附錄 82

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