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研究生:陳詩馨
研究生(外文):Shih-Shin Chen
論文名稱:薑黃素抑制皮膚黑色素細胞生成黑色素的效果與在化妝品的應用
論文名稱(外文):The inhibition effect of curcumin on the melanin formation by melanocytes and the application to cosmetics
指導教授:陳榮輝陳榮輝引用關係
指導教授(外文):Rong-Heui Chen
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
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:89
中文關鍵詞:薑黃素黑色素化妝品
外文關鍵詞:curcuminmelanincosmetics
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摘要
本論文的研究目的在探討薑黃素 (curcumin) 抑制皮膚黑色素細胞生成黑色素的效果與其在化妝品上的應用。實驗分為三部份:第一部份在探討薑黃素對皮膚的美白效果,焦點集中在薑黃素對人類黑色素腫瘤細胞 (A375) 存活率的影響與對人類黑色素腫瘤細胞 (A375) 之黑色素生成量的抑制效果;第二部份在探討薑黃素對皮膚的抗老化效果,包括對纖維母細胞 (CCD-966SK) 增生的效果、對促進膠原蛋白 (collagen) 分泌的效果、薑黃素的還原力與捕捉DPPH自由基的抗氧化能力;第三部份在探討薑黃素製成乳霜後,產品的安定性、安全性及對皮膚特性改良的有效性。
實驗的結果如下:
薑黃素於濃度 0.09 mM 時對黑色素腫瘤細胞 A375 的存活開始產生抑制現象,在薑黃素濃度達 0.24 mM 時,其存活率降至30%以下;於薑黃素濃度 4.68 x 10-4 至 0.24 mM 之間時,A375 經UVA照射後所產生的黑色素明顯減少,抑制率為 19% - 67% ;於薑黃素濃度 4.68 x 10-4 至 0.24 mM 之間時,A375經UVB照射後所產生的黑色素減少得更多,抑制率提高為 52% - 72%。
薑黃素在濃度 4.68 x 10-4 至 0.24 mM 之間時對人類皮膚纖維母細胞 CCD-966SK 的存活率皆維持在 86% ~ 101%,顯示薑黃素對皮膚纖維母細胞並無明顯的抑制或增生效果;在薑黃素濃度為 3.75 x 10-3 mM 和 0.015 mM時,皮膚膠原蛋白總生成量達到與控制組相同的29 μg,而在其它濃度下的皮膚膠原蛋白生成量亦相差無幾,可知薑黃素對皮膚膠原蛋白也無明顯的促生效果。
薑黃素的還原力遠高於維生素 C 的還原力;其清除 DPPH 自由基的能力也均接近 100%,相較於維生素 C 的表現僅有些微的落後。這些結果顯示薑黃素具有優異的抗氧化能力。
含薑黃素的乳霜經 135 W,42 KHz 超音波震盪 5 分鐘、3000 rpm 離心 30 分鐘後不會發生相分離;在 50℃ 中貯存 2 個月後也不會發生變質的現象,顯示出含薑黃素的乳霜具有良好的安定性。
含薑黃素的乳霜,使用六週後測得人體pH值為5.24-5.98 且不會有皮膚紅腫的現象,顯示出該產品具有良好的安全性。
含薑黃素的乳霜對皮膚保水性無影響;塗擦含薑黃素的乳霜六週後,對實驗組皮膚的 R2 彈性值、 R8 彈性值和鱗片度都有統計上的顯著提昇,而平滑度和粗糙度雖然實驗組高於控制組,但是並未達到統計上的顯著差異性;塗擦含薑黃素的乳霜六週後,黑色素指標出現降低的現象,代表薑黃素具有皮膚美白的效果。
Abstract
The aim of this research is to explore the whitening and anti-aging effects of cosmetics containing curcumin. The study includes three parts: 1. to evaluate the viability of human malignant melanoma (A375); skin whitening effect of curcumin in terms of the inhibition of melanin formation by human malignant melanoma (A375); 2. anti-aging effect of curcumin in terms of the fibroblasts (CCD-966SK) proliferation, collagen excretion, reducing power and anti-oxidation ability based on DPPH scavenging rate; 3. stability, safety and efficacy characteristics of cream containing curcumin.
The results obtained are: with concentrations of 0.09 mM or higher, curcumin shows inhibition on viability of A375. The cell viability decreases to below 30% at 0.24 mM of curcumin. With concentrations between 4.69 x 10-4 and 0.24 mM, curcumin shows 19% - 67% inhibition on melanin formation by A375 after UVA radiation (1.1 mW/cm2, 15 mins), while 52% - 72% inhibition on that after UVB radiation (1.5 mW/cm2, 15 mins).
Between 4.68 x 10-4 to 0.24 mM of curcumin, the cell viabilities of skin fibroblasts are among 86% ~ 100%. It indicates that curcumin has neither inhibition nor proliferation effects on the skin fibroblasts proliferation. With the concentrations of curcumin at 3.75 x 10-3 mM and 0.015 mM, the excretion of collagen by fibroblasts is around 29 μg.
At all concentrations, curcumin shows much better reducing power than Vit. C. In addition, all the DPPH scavenging rates of curcumin exceed 100%, slightly lower than that of Vit. C. These mean that curcumin has outstanding ability of anti-oxidation.
The stability of cream products containing curcumin is assessed for sonication (135 W, 42KHz, 5 mins), for centrifuging (3000 rpm, 30 mins) , and for more than two months’ high temperature (50□C) storage. The cream products remain stable without phase separation. The pHs of cream products have no significant difference after sonication or two months’ high temperature storage. These mean that cream products containing curcumin have good stability.
After applying the cream products containing curcumin for six weeks, no erythema can be observed. The pHs of cream products are between 5.24-5.98 during that time. These mean that cream products containing curcumin have good safety.
Curcumin shows no improvments on water-retaining capability. After applying the cream products containing curcumin for six weeks, the elasticities R2, R8 and scaliness decrease ratios in treatment group all possess significant progress, while the elasticities R2, R8, scaliness decrease ratios, wrinkles decrease ratios and water-retaining capability all appear significant differences between the treatment group and the control one. After applying the cream products containing curcumin for six weeks, the melanin index decreases. It means curcumin has skin whitening effects.
目錄
中文摘要………………………………………………………………ⅵ
英文摘要………………………………………………………………ⅷ
圖目錄…………………………………………………………………ⅹ
表目錄…………………………………………………………………ⅺ
壹、前言………………………………………………………………1
貳、文獻整理…………………………………………………………3
一、化妝品產業之趨勢與特色………………………………………3
二、薑黃素之簡介……………………………………………………3
1.薑黃的植物學特徵與成分…………………………………………4
2.薑黃素的物理性質與化學性質……………………………………5
3.薑黃素的代謝………………………………………………………5
4.薑黃素的生理活性…………………………………………………6
5.薑黃素的不穩定性…………………………………………………8
三、皮膚老化的生理機能變化………………………………………9
1.皮膚的生理功能……………………………………………………9
2.皮膚的結構…………………………………………………………9
3.皮膚老化的原因……………………………………………………12
4.皮膚老化的自由基學說……………………………………………12
四、抗氧化作用………………………………………………………13
1.自由基與活性含氧族群……………………………………………13
2.抗氧化系統…………………………………………………………14
3.紫外線對於皮膚抗氧化系統之影響………………………………17
五、皮膚黑色素形成…………………………………………………17
1.黑色素細胞…………………………………………………………17
2.黑色素的形成機轉…………………………………………………18
3.黑色素之功用………………………………………………………20
4.酪胺酸酶的性質……………………………………………………21
六、美白的方法………………………………………………………21
1.中醫美白理論………………………………………………………21
2.西醫美白理論………………………………………………………22
七、乳霜之應用………………………………………………………23
1.乳霜的定義…………………………………………………………23
2.乳霜的分類…………………………………………………………23
參、材料與方法………………………………………………………25
一、實驗大綱…………………………………………………………25
二、實驗流程…………………………………………………………25
三、實驗材料…………………………………………………………26
1.細胞株………………………………………………………………26
2.藥材…………………………………………………………………26
3.藥品…………………………………………………………………26
4.設備…………………………………………………………………28
四、實驗方法…………………………………………………………29
1.細胞培養實驗………………………………………………………29
1.1 細胞陪養…………………………………………………………29
1.2 細胞繼代培養……………………………………………………30
1.3 細胞解凍與保存…………………………………………………30
1.4 細胞存活率………………………………………………………31
2.薑黃素美白效果的細胞株試驗……………………………………32
2.1 薑黃素對人類黑色素腫瘤細胞存活率之影響…………………32
2.2 薑黃素對抑制人類黑色素腫瘤細胞生成黑色素之影響………32
3.薑黃素抗老化功效之抗氧化試驗及細胞株試驗…………………32
3.1 還原力之測定……………………………………………………32
3.2 清除DPPH自由基能力之測定……………………………………33
3.3 薑黃素對人類纖維母細胞存活率之影響………………………34
3.4 薑黃素對膠原蛋白合成之影響…………………………………34
4.含薑黃素之乳霜之製作方法………………………………………35
5.含薑黃素乳霜之品質試驗…………………………………………35
5.1 色度測試…………………………………………………………36
5.2 安定性測試………………………………………………………36
5.3 安全性測試………………………………………………………37
5.4 有效性測試………………………………………………………37
6.統計分析……………………………………………………………39
肆、結果與討論………………………………………………………40
1.評估薑黃素美白功效之細胞株試驗………………………………40
1.1 薑黃素對於人類黑色素腫瘤細胞存活率之影響………………40
1.2 薑黃素對於抑制人類黑色素腫瘤細胞生成黑色素之影響……40
2.評估薑黃素抗老化功效之細胞株試驗及抗氧化試驗……………41
2.1 薑黃素對人類纖維母細胞存活率之影響………………………41
2.2 薑黃素對膠原蛋白增生之影響…………………………………42
2.3 薑黃素還原力之測定……………………………………………43
2.4 薑黃素清除DPPH自由基之能力…………………………………43
3.含薑黃素乳霜之品質分析…………………………………………43
3.1 含薑黃素之乳霜之色度…………………………………………43
3.2 含薑黃素乳霜之物理安定性……………………………………44
3.3 含薑黃素乳霜對皮膚之安全性…………………………………44
4.含薑黃素乳霜對皮膚有效性之測試………………………………45
4.1 含薑黃素乳霜對皮膚之保水性…………………………………45
4.2 含薑黃素乳霜對皮膚之彈性……………………………………46
4.3 含薑黃素乳霜對皮膚之美白……………………………………47
4.4 含薑黃素乳霜對皮膚之粗糙度…………………………………47
伍、結論………………………………………………………………50
陸、參考文獻…………………………………………………………52
圖………………………………………………………………………61
表………………………………………………………………………77
附錄……………………………………………………………………81
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