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研究生:謝詠筌
研究生(外文):Yung - Chuan Hsieh
論文名稱:建立真菌材料於化妝品之功效篩選技術
論文名稱(外文):Establish an evaluation system for fungal materials using in cosmetic.
指導教授:陳啟楨陳啟楨引用關係陳健祺
指導教授(外文):Chee - Jen ChenJian - Chi Chen
學位類別:碩士
校院名稱:南台科技大學
系所名稱:生物科技系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:105
中文關鍵詞:真菌化妝品白木耳黑木耳紅麴酒粕酪胺酸酶黑色素基質金屬蛋白酶一氧化氮美白抗皮膚光老化抗氧化抗發炎
外文關鍵詞:fungi、cosmetic、Tremella、 Auricularia polytricha、Monascus purpureus、tyrosinase、melanin、MMPs、nitric oxide、DCFH-D
相關次數:
  • 被引用被引用:4
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  • 下載下載:220
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亞洲人光老化的皮膚臨床特徵為色素沉積和出現皺紋。本研究中以真菌材料為試驗對象;Thanaka粉末不屬於真菌材料範疇內,在此做為評估比較用。試驗材料分為(A)、(B)、(C)三類;(A)為六種白木耳菌株液態發酵液冷凍乾燥粉末、(B)為六種白木耳菌株液態發酵液酒精沉澱之粗多醣冷凍乾燥粉末、(C)為三種樣品粉末之甲醇萃取物和酒精萃取物冷凍乾燥粉末。
六種白木耳菌株分別為: T. encephala T897、T. flava CCJ00907、T. nivalis CCJ00911、T. flava CCJ00928、T. sp. CCJ00933和T. fucifomis CCJ00960。(C)類三種樣品粉末分別為: Thanaka粉末、黑木耳粉末、紅麴酒粕粉末。
數據結果顯示500μg/mL T. fuciformis液態發酵液冷凍乾燥粉末F960、T. flava發酵液之粗多醣冷凍乾燥粉末P928和Thanaka酒精萃取物冷凍乾燥粉末TPEE具有抑制B16F10小鼠黑色素瘤細胞之tyrosinase活性和黑色素生合成。
Gelatin zymography試驗中發現100μg/mL T. sp.發酵液之粗多醣冷凍乾燥粉末P933 、T. fuciformis發酵液之粗多醣冷凍乾燥粉末P960 、黑木耳粉末甲醇萃取物冷凍乾燥粉末APME和紅麴酒粕甲醇萃取物冷凍乾燥粉末MPME皆能有效地減少MMP ( matrix metalloprotease )分解明膠之活性及抑制MMP基因轉錄;這些原料可朝向改善光老化作用方面發展。
細胞氧化傷害保護試驗和小鼠正常纖維母細胞增生試驗之結果相互比對下,發現PDB液態發酵液冷凍乾燥粉末Pb、T. encephal發酵液之粗多醣冷凍乾燥粉末P897、T. flava發酵液之粗多醣冷凍乾燥粉末P907 、黑木耳粉末甲醇萃取物冷凍乾燥粉末APME、黑木耳粉末酒精萃取物冷凍乾燥粉末APEE 和紅麴酒粕甲醇萃取物冷凍乾燥粉末MPME可保護對抗過氧化氫對NIH 3T3老鼠正常纖維母細胞所造成之細胞毒性,其作用可能由於對小鼠正常纖維母細胞具有增生作用所導致。而細胞內活性氧物質含量試驗結果發現500μg/mL F911 T. nivalis液態發酵液冷凍乾燥粉末、T. flava發酵液之粗多醣冷凍乾燥粉末P907可減少經由過氧化氫所誘導之活性氧物質產生。
一氧化氮含量測定試驗結果發現100μg/mL Thanaka甲醇萃取物冷凍乾燥粉末TPME、Thanaka酒精萃取物冷凍乾燥粉末TPEE、黑木耳粉末甲醇萃取物冷凍乾燥粉末APME 和黑木耳粉末酒精萃取物冷凍乾燥粉末APEE可抑制經由脂多醣 (lipopolysaccharide, LPS) 誘導之一氧化氮生成;但對於在基因轉錄時期抑制iNOS表現之效果並不明顯。
本研究為找尋可有效應用於化妝品方面之生理活性物質,建立應用於美白、阻止皮膚光老化、抗氧化、抗發炎之體外試驗篩選評估技術平台。
The clinical characteristics of photoaging in Asian skin, such as pigmentary changes and wrinkle patterns. The freeze-dried powder of submerged culture of six Tremella strains (A), and the freeze-dried powder of polysaccharide extracted from submerged culture of six Tremella strains (B), and the freeze-dried powder of methanol extracts and ethanol extracts from three different substrates (C) were compared and its character were explored in this study.
The six Tremella stains: T. encephala T897、T. flava CCJ00907、T. nivalis CCJ00911、T. flava CCJ00928、T. sp. CCJ00933 and T. fucifomis CCJ00960. The three different substrates: Thanaka powder、Auricularia polytricha powder、Monascus purpureus vinasse powder.
These data demonstrate that 500μg/mL of F960、P928 and TPEE inhibit the tyrosinase activity and down-regulates melanogenesis significantly in B16F10 mouse melanoma cells.
In particular, 100μg/mL of P933、P960、APME and MPME are effective in reducing the gelatinase activities and MMP gene transcription. These results suggest that specific inhibition of MMP activity immediately after sun exposure could provide an effective way of preventing sun-induced damage of dermal extracellular matrix. Therefore, It can be developed as a promising photoaging-improving topical agent.
Moreover, the results demonstrate that 500μg/mL of Pb、P897、P907、APME、APEE and MPME decrease the cyto-toxicity of hydrogen peroxide, the effect may be by the notable proliferation activity for NIH 3T3 mouse fibroblasts.
Scavenging activity of reactive oxygen species (ROS) was evaluated by fluorescent probe DCFH-DA flow cytometry. F911 and P907 were proved inhibit the hydrogen peroxide-induced release of reactive oxygen species.
Results indicate 100μg/mL of TPME、TPEE、APME and APEE inhibit the production of nitric oxide by lipopolysaccharide-induced RAW 264.7 macrophages, but do not affect the mRNA levels of inducible nitric oxide synthase (iNOS).The results suggest that those materials reduced inflammatory mediators and may be a powerful agent to inhibit inflammatory erythema in human skin.
This study tried to establish an evaluation system on whitening agents, improvement of photoaged skin, anti-oxidants, and anti-inflammatory activity using in cosmetic.
目次 Ⅰ
表目錄 Ⅳ
圖目錄 Ⅴ
中文摘要 Ⅶ
英文摘要 Ⅷ
縮寫表 Ⅸ
第一章 緒論……………………………………………………………1
第二章 文獻整理………………………………………………………3
2.1 皮膚結構……………………………………………………………3
2.2 紫外線照射對皮膚之影響…………………………………………5
2.2.1 紫外線對皮膚之毒性效應…………………………………………5
2.2.2 紫外線照射之光致癌性……………………………………………6
2.2.3 紫外線照射引發黑色素生合成……………………………………6
2.3 黑色素生合成作用及其調節機轉…………………………………7
2.3.1 紫外線對黑色素合成之調控………………………………………9
2.3.2 內分泌對黑色素合成之調控………………………………………9
2.3.3 防止或抑制黑色素形成之機制……………………………………10
2.4 皮膚光老化…………………………………………………………12
2.4.1 UV照射造成皮膚光老化機制………………………………………13
2.4.2 基質金屬蛋白酶與皮膚光老化之關連性…………………………14
2.5 皮膚與一氧化氮關係………………………………………………21
2.6 試驗材料介紹………………………………………………………23
2.7 實驗架構……………………………………………………………27
第三章 材料與方法……………………………………………………28
3.1 實驗儀器與藥品……………………………………………………28
3.2 試驗材料製備………………………………………………………30
3.3 實驗方法……………………………………………………………33
3.3.1 細胞毒性試驗………………………………………………………33
3.3.2 Tyrosinase hydroxylasedase活性試驗……………………34
3.3.3 Dopa oxidase活性試驗…………………………………………35
3.3.4 Tyrosinase zymography活性試驗……………………………36
3.3.5 細胞內黑色素含量試驗……………………………………………38
3.3.6 α-MSH誘導黑色素生成試驗活性試驗……………………………39
3.3.7 Gelatin zymography活性試驗…………………………………39
3.3.8 MMP-2及MMP-9 mRNA表現量試驗…………………………………42
3.3.9 小鼠正常纖維母細胞增生試驗……………………………………43
3.3.10 細胞氧化傷害保護試驗……………………………………………44
3.3.11 細胞內活性氧含量試驗……………………………………………44
3.3.12 一氧化氮生成試驗…………………………………………………46
3.3.13 iNOS mRNA表現量試驗……………………………………………46
3.4 統計方法………………………………………………………………47
第四章 實驗結果….……………………………………………………48
4.1 受測樣品經冷凍乾燥之產率…………………………………………48
4.2 細胞毒性試驗…………………………………………………………48
4.3 Tyrosinase hydroxylasedase活性試驗………………………48
4.4 Dopa oxidase活性試驗……………………………………………49
4.5 Tyrosinase zymography活性試驗………………………………49
4.6 細胞內黑色素含量試驗………………………………………………49
4.7 α-MSH誘導黑色素生成試驗活性試驗………………………………50
4.8 Gelatin zymography活性試驗……………………………………51
4.9 MMP-2及MMP-9 mRNA表現量試驗……………………………………51
4.10 小鼠正常纖維母細胞增生試驗………………………………………51
4.11 細胞氧化傷害保護試驗………………………………………………51
4.12 細胞內活性氧含量試驗………………………………………………52
4.13 一氧化氮生成試驗……………………………………………………52
4.14 iNOS mRNA表現量試驗………………………………………………52
第五章 討論………………………………………………………………53
5.1 細胞毒性試驗…………………………………………………………53
5.2 Tyrosinase hydroxylasedase活性試驗………………………53
5.3 Dopa oxidase活性試驗……………………………………………53
5.4 Tyrosinase zymography活性試驗………………………………54
5.5 細胞內黑色素含量試驗………………………………………………54
5.6 α-MSH誘導黑色素生成試驗活性試驗………………………………55
5.7 Gelatin zymography活性試驗……………………………………56
5.8 MMP-2及MMP-9 mRNA表現量試驗……………………………………56
5.9 小鼠正常纖維母細胞增生試驗………………………………………56
5.10 細胞氧化傷害保護試驗………………………………………………57
5.11 細胞內活性氧含量試驗………………………………………………57
5.12 一氧化氮生成試驗……………………………………………………58
5.13 iNOS mRNA表現量試驗………………………………………………59
5.14 試驗結果討論…………………………………………………………59
第六章 總結.………………………………………………………………60
第七章 參考文獻.…………………………………………………………62
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