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研究生:吳佳峻
研究生(外文):WU, CHIA-CHUN
論文名稱:白楊素藉由下調血管內皮生長因子受器抑制高糖誘導脈絡膜內皮細胞之細胞遷移
論文名稱(外文):Chrysin Inhibits High Glucose-Induced Migration on Choroidal Endothelial Cells via VEGF Receptor Down-regulation
指導教授:洪啟峯
指導教授(外文):HUNG, CHI-FENG
口試委員:王素珍黃聰龍方嘉佑
口試委員(外文):WANG, SU-JANEHWANG, TSONG-LONGFANG, JIA-YOU
口試日期:2017-07-24
學位類別:碩士
校院名稱:輔仁大學
系所名稱:生物醫學暨藥學研究所碩士班
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:103
中文關鍵詞:高糖白楊素脈絡膜內皮細胞血管內皮生長因子
外文關鍵詞:high glucosechrysinchoroidal endothelial cellvascular endothelial growth factor (VEGF)
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糖尿病是屬於一種慢性發炎疾病且經常伴隨許多併發症的產生,如糖尿病視網膜病變,是造成糖尿病病患眼睛受損的主要原因之一。在先前的諸多文獻中皆指出白楊素具有許多生物活性如抗氧化、抗發炎等特性。然而,白楊素在眼睛疾病上之應用文獻仍未普遍。因此,在本篇研究中,探討白楊素對於高糖誘導脈絡膜內皮細胞 (RF/6A) 遷移之抑制效果及其機制作用為何。在研究結果中發現,透過Transwell及傷口癒合之實驗方法,證實白楊素具有能夠抑制由高糖誘導的脈絡膜內皮細胞遷移之特性且效果與劑量呈正相關。在細胞存活率實驗中,觀察到不同濃度的白楊素對脈絡膜內皮細胞皆無細胞毒殺之特性。此外,白楊素抑制細胞遷移之分子機制,透過下調蛋白激酶B (AKT) 及細胞外訊息調節激酶 (ERK) 磷酸化,以及降低基質金屬蛋白酶-2 (MMP-2) 之表現量。白楊素也減少血管內皮生長因子之轉錄因子表現量,但血管內皮生長因子其表現量不受白楊素影響。進一步發現,白楊素抑制血管內皮生長因子受器之蛋白質及mRNA表現量。這些結果證明,白楊素可抑制由高糖誘導脈絡膜內皮細胞之細胞遷移進而達到保護效果。
Diabetes mellitus (DM) is a chronic inflammatory disease and usually accompanies with some complications such as diabetic retinopathy (DR), which is a major cause of visual impairment for diabetic patients. Numerous studies have shown that chrysin, one of flavonoids, has many biological activities such as anti-oxidation and anti-inflammation, etc. However, it is rarely used in ocular diseases. In this study, we examined the inhibitory effects of flavonoid on high glucose induced migration of RF/6A cell and its mechanism. The result shown that chrysin can inhibit the RF/6A cell migration which is induced by high glucose with dose dependent manner in transwell assay and scratch assay. Treatment of chrysin on RF/6A cell at different concentrations was found non-cytotoxicity in MTT assay. Moreover, chrysin down-regulated both phosphorylated AKT and ERK, as well as attenuated the expression levels of MMP-2. It also decreased the expression of VEGF transcription factor, but not effect on the expression of VEGF. Furthermore, chrysin could suppress the protein and mRNA expression levels of VEGF receptor. These results indicated that chrysin can suppress migration of high glucose-induced RE/6A cell, and it might be have the function of visual protection.
摘要 II
Abstract IV

第一章 緒言 1
第二章 文獻回顧 3
第一節 類黃酮之功用 3
第二節 糖尿病 5
第三節 糖尿病併發症 12
第四節 血管新生及血管內皮生成因子 15
第五節 眼睛結構 22
第三章 研究動機與目的 35
第四章 實驗材料與器材 36
第一節 儀器 36
第二節 材料 37
第五章 研究方法 38
一、 細胞培養 38
二、 白楊素給予方法及高糖誘導方式 39
三、 細胞遷移試驗 (Transwell assay) 39
四、 傷口癒合試驗 (Scratch wound healing assay) 40
五、 細胞存活率測試 (MTT assay) 41
六、 西方墨點法分析試驗 (Western blotting analysis) 42
七、 酵素免疫分析法 (Enzyme-linked immunosorbent assay, ELISA) 44
八、 即時聚合酶鏈式反應 (Quantitative real time polymerase chain reaction, RT-qPCR) 46
九、 統計方法 48
第六章 實驗結果 50
一、 白楊素抑制高糖誘導脈絡膜內皮細胞之細胞遷移 50
二、 白楊素對於脈絡膜內皮細胞無細胞毒性 51
三、 白楊素抑制細胞遷移透過抑制AKT及ERK磷酸化及降低MMP-2表現量 52
四、 白楊素抑制血管內皮生長因子之轉錄因子表現量但不影響血管內皮生長因子表現量 53
五、 白楊素下調血管內皮生長因子受器蛋白質及mRNA表現量 54
第七章 討論 56
第八章 結論 61
參考文獻 85


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