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研究生:周佳蓉
研究生(外文):Chia-Jung Chou
論文名稱:含羞草對紫外線照射人類纖維母細胞的保護作用
論文名稱(外文):PROTECTIVE EFFECTS OF MIMOSA PUDICA AGAINST UVB-INDUCED DAMAGE OF HUMAN FIBROBLASTS
指導教授:李綉鈴
指導教授(外文):Shiow-Ling Lee
口試委員:李綉鈴
口試委員(外文):Shiow-Ling Lee
口試日期:2015-01-29
學位類別:碩士
校院名稱:大同大學
系所名稱:生物工程學系(所)
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:86
中文關鍵詞:紫外線光老化含羞草
外文關鍵詞:photoagingM. pudica
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紫外線輻射刺激皮膚產生氧化壓力,造成皮膚光老化。具有抗光老化之中草藥近年來逐漸受到重視。本研究評估富含多酚化合物之含羞草對經紫外線照射後之人類纖維母細胞CCD966SK的保護作用。結果顯示,含羞草對CCD966SK細胞毒性很低,100 ?慊/ml甲醇萃取物不會降低細胞存活率。以UVB照射CCD966SK細胞,細胞存活率則隨著照射劑量增加而顯著下降;若先以含羞草甲醇或熱水萃取物與CCD966SK細胞共培養24小時後,再照射UVB25 mJ/cm2或50 mJ/cm2,則能增加細胞存活率與膠原蛋白生成,且隨著萃取物濃度之增加而增加,其中以100 ?慊/ml含羞草甲醇萃取物處理組效果最佳,與正控制組(100 ?慊/ml抗壞血酸)無顯著之差異,顯示含羞草可增加經UVB照射的纖維母細胞存活率與膠原蛋白生成。此外,經UVB照射之CCD966SK細胞,其榖胱苷肽含量下降,而脂質過氧化物丙二醛含量則明顯上升。而以含羞草萃取物共培養的CCD966SK細胞,再照射UVB,則能提升經UVB照射CCD966SK細胞之榖胱苷肽含量,並降低因UVB照射增加的丙二醛含量,在25及50 mJ/cm2 UVB之照射劑量下,甲醇萃取物處理效果顯著優於熱水萃取物處理者。這些結果顯示含羞草具有降低UVB照射纖維母細胞的傷害。
Ultraviolet radiation of skin resulted in oxidative stress, causing photoaging of skin. Recently the anti-photoaging property of herbs had attracted much attention. The protective effects of Mimosa pudica against UVB-induced damage of human fibroblasts CCD966SK was determined in this study. The results showed that 100 ?慊/ml of methanol extract of M. pudica did not reduce the cell viability, indicating low cytotoxicity of M. pudica to CCD966SK cells. A dose-dependent decrease in cell viability was found in UV radiated CCD966SK cells. However, cell viability and collagen formation increased in CCD966SK cells pretreated with increasing methanol and water extracts for 24 h, respectively, followed by UVB radiation. Methanol extracts of 100 ?慊/ml exhibited the maximum effect without statistically significant difference to ascorbic acid as a positive control. This demonstrated that M. pudica increased the viability and collagen production in UVB-exposed CCD966SK cells. A significant decrease in glutathione and increase in malondialdehyde were obtained in CCD966SK cells with UVB irradiation. Pretreatment of M. pudica also promoted glutathione and reduced malondialdehyde amounts in CCD966SK cells with UVB exposure. Methanol extract had significant better effect than water extract on the CCD966SK cells exposed at 25 and 50 mJ/cm2. These results indicated that M. pudica could reduce the UVB-induced damage in human dermal fibroblasts.
目錄
誌謝 I
摘要 II
英文摘要 III
目錄 V
圖目錄 IX
表目錄 XI
壹、前言 1
貳、文獻回顧 2
一、皮膚的功能與結構 2
(一)表皮層(epidermis): 2
(二)真皮層(dermis): 2
(三)皮下組織(subcutaneous tissue) : 5
二、基質金屬蛋白酶 5
三、皮膚老化 7
(一)內因性老化 7
(二)外因性老化 7
四、太陽光 8
(一)紫外線 11
(二)紫外線輻射對皮膚的影響 11
(三)皮膚癌 19
五、自由基的介紹 20
(一)自由基來源 20
(二)自由基定義和種類 20
六、抗氧化物質 21
(一)維生素系統 21
(二)酵素系統 22
七、光保護 26
(一) 綠茶(green tea) 26
(二) 水飛薊(silibinin) 26
(三)黃豆(Glycine max L.) 27
(四)大蒜(Allium sativum L.) 27
(五)月見草(Oenothera laciniata) 28
八、含羞草介紹 28
(一)含羞草之傳統藥理功用 28
(二)含羞草之現代科學藥理功用 29
(三) 前人發表之含羞草相關研究 29
參、材料與方法 31
一、實驗架構 31
二、材料與方法 32
(一)實驗樣品 32
(二)實驗藥品 33
(三)實驗儀器 34
三、實驗方法 35
(一)含羞草不同溶劑萃取之製備 35
(二)CCD966SK細胞之活化 36
(三)細胞接種 37
(四)含羞草對CCD966SK細胞之毒性試驗 37
(五)紫外線處理CCD966SK細胞 40
(六)含羞草對經紫外線照射CCD966SK細胞之抗氧化相關參數及膠原蛋白含量之影響 42
(七)數據之統計分析 47
肆、結果與討論 48
一、含羞草不同溶劑萃取物對CCD966SK細胞之毒性試驗 48
二、UVB對CCD966SK細胞之傷害 52
三、含羞草萃取物對經UVB照射之CCD966SK細胞存活率 55
四、含羞草不同溶劑萃取物對CCD966SK細胞膠原蛋白生成 59
五、含羞草萃取物對經UVB照射CCD966SK胞內膠原蛋白生成 63
六、含羞草萃取物對經UVB照射之CCD966SK細胞MDA生成 67
七、含羞草萃取物對經UVB照射CCD966SK細胞之GSH含量 71
伍、結論 76
陸、參考文獻 77
柒、附錄 88


圖目錄
圖2.1皮膚的構造 3
圖2.2 UV照射皮膚導致老化 9
圖2.3太陽輻射光譜 10
圖2.4紫外線分類 12
圖2.5紫外線照射皮膚之影響 14
圖2.6 ROS在光老化的作用 15
圖2.8 MAPK訊息傳遞路徑 18
圖2.9 在皮膚細胞之ROS生成與抗氧化物防禦 23
圖3.1實驗架構流程圖 31
圖3.2從tetrazolium salt與SDH反應生成Formazan 39
圖3.3 MDA與TBA反應生成MDA-TBA2 43
圖3.4 GSH-OPA反應示意圖 45
圖4.2不同強度之UVB對CCD966SK細胞存活率之影響。 53
圖4.3含羞草萃取物對經UVB照射後CCD966SK細胞存活率 56
圖4.4含羞草不同溶劑萃取物對CCD966SK細胞膠原蛋白生成 60
圖4.5含羞草萃取物對經UVB照射之CCD966SK細胞膠原蛋白生成 64
圖4.6含羞草萃取物對經UVB照射之CCD966SK細胞MDA含量 68
圖4.7含羞草萃取物對經UVB照射之CCD966SK細胞GSH含量 73
圖7.1 Hydroxyproline assay之檢量線 88


表目錄
表2.1基質金屬蛋白酶 6
表4.1含羞草不同溶劑萃取物對CCD966SK細胞存活率 50
表4.2含羞草萃取物對經UVB照射之CCD966SK細胞存活率 57
表4.3含羞草不同溶劑萃取物對CCD966Sk細胞膠原蛋白生成 61
表4.4含羞草萃取物對經UVB照射之CCD966SK細胞膠原蛋白生成 65
表4.5含羞草萃取物對經UVB照射之CCD966SK細胞MDA生成 70
表4.6含羞草萃取物對經UVB照射之CCD966SK細胞GSH含量 75
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