TW202137976A - 包含植物萃取物的組成物及其修護受損皮膚纖維母細胞之用途 - Google Patents
包含植物萃取物的組成物及其修護受損皮膚纖維母細胞之用途 Download PDFInfo
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Abstract
本發明係關於一種用於修護受損的纖維母細胞的植物萃取物的組成物。該組成物係由紅茶、綠茶、普洱茶、四季春茶、紅酒、藍莓或葡萄籽其中之一與柑橘的萃取物組合所組成。
Description
本發明係關於一種包含植物萃取物的組成物及其用途，特別係關於一種減少紫外線所致皮膚損傷之組成物及其用途。
太陽是大多數人接受紫外線(ultraviolet，UV)照射的主要來源。紫外線可細分為紫外線A (UVA；波長315–400 nm)、紫外線B (UVB；280–315 nm)、及紫外線C (UVC；100–280 nm)。紫外線C被大氣吸收而不會到達地球表面。紫外線B是對生物造成最嚴重損傷的紫外線，然其大部分被大氣吸收。長波長的紫外線A是到達地球的紫外線中強度最高者，其可穿透身體組織深處。研究發現暴露於紫外線下有較高機率加速皮膚老化與罹患皮膚癌，並且可能降低個人對傳染性疾病抵抗力。
皮膚是成人中面積超過1.5 m2
的大型器官，它提供了對抗陽光中紫外線輻射的第一階段保護。皮膚包含表皮(包括最外側的角質層)、真皮、及皮下組織。表皮是皮膚的最外層並且不斷更新。表皮與真皮間存在持續分裂的細胞(如纖維母細胞、角質細胞、黑色素細胞)，該些細胞的活動對紫外線非常敏感。真皮含有膠原蛋白與彈性蛋白，其賦予肌膚彈性和支撐力量。皮膚透過增加黑色素的生成及促進細胞增生以增厚皮膚外層(角質層)等方式阻止紫外線輻射對表皮以下組織造成的損傷。例如，當暴露於高量的紫外線(主要是紫外線A)，膠原纖維與彈性蛋白會受損，因而造成皮膚彈性降低、皺紋形成、及出現皮膚老化的外觀。
減少紫外線照射所致損傷最普遍的方法是避免陽光曝曬及使用防曬品。為獲得足夠的保護，應將防曬品大範圍地塗抹於皮膚及確保其厚度均勻，並在外出前讓其乾燥約15分鐘。此外，間隔一段時間需要再次塗抹防曬品以維持其厚度。多數人由於未依照前述方法使用防曬品而無法獲得良好的保護。在某些情況下，防曬品含有的化學物質可能引發光敏性，以致該些化學物質與紫外線的結合對皮膚產生不良影響，例如皮疹或更嚴重的曬傷。
有鑑於此，開發一種成分天然又能有效保護皮膚免於紫外線傷害的組成物，以減緩皮膚老化或降低皮膚癌的發生率，實有其必要。
緣此，本發明之一目的在提供一種減少紫外線所致皮膚損傷的組成物，係選自由菠菜萃取物與紅茶萃取物、菠菜萃取物與紅酒萃取物、菠菜萃取物與藍莓萃取物、菠菜萃取物與葡萄籽萃取物、柑橘萃取物與紅茶萃取物、柑橘萃取物與綠茶萃取物、柑橘萃取物與普洱茶萃取物、柑橘萃取物與四季春茶萃取物、柑橘萃取物與紅酒萃取物、柑橘萃取物與青咖啡豆萃取物、柑橘萃取物與菠菜萃取物、及柑橘萃取物與葡萄籽萃取物所組成群組中任一組合所組成。
在本發明之一實施例中，該包含菠菜萃取物之組成物係由至少0.078125 mg/ml之菠菜萃取物，及至少0.078125 mg/ml之紅茶萃取物、紅酒萃取物、藍莓萃取物、或葡萄籽萃取物所組成。
在本發明之一實施例中，該包含柑橘萃取物之組成物係由至少0.015625 mg/ml之柑橘萃取物，及至少0.015625 mg/ml之紅茶萃取物、綠茶萃取物、普洱茶萃取物、四季春茶萃取物、紅酒萃取物、青咖啡豆萃取物、菠菜萃取物、或葡萄籽萃取物所組成。
本發明之另一目的在提供一種減少紫外線所致皮膚損傷之醫藥組成物，包含前述任一種組成物，及一藥學上可接受之載劑。
在本發明之一實施例中，前述醫藥組成物之劑型可為溶液、粉末、膠囊、或錠劑。
本發明之又一目的在提供前述任一種組成物用於減少紫外線所致皮膚纖維母細胞之損傷之用途，或者前述任一種組成物用於製備減少紫外線所致皮膚損傷之醫藥組成物。
在本發明之一實施例中，該紫外線係為紫外線A。
本發明之組成物藉由混合特定植物萃取物大幅抑制紫外線照射所致皮膚纖維母細胞死亡。皮膚纖維母細胞的正常增生則有利於維持表皮的厚度及紫外線屏障功能，亦可補充皮膚細胞外基質中的組成及維持真皮的更新。換言之，對一個體施予有效量之本發明組成物顯著減少該個體中紫外線所致皮膚損傷。因此，本發明之組成物可用於製備保護皮膚對抗紫外線所致損傷的醫藥組成物。
以下將配合圖式進一步說明本發明的實施方式，下述所列舉的實施例係用以闡明本發明之發明特點及應用，而非以限定本發明之範圍，任何熟習此技藝者，在不脫離本發明之精神和範圍內，當可做些許更動與潤飾，因此本發明之保護範圍當視後附之申請專利範圍所界定者為準。
定義
本文中所使用數值為近似值，所有實驗數據皆表示在20%的範圍內，較佳為在10%的範圍內，最佳為在5%的範圍內。
本文中所謂「藥學上可接受之載劑」指一種或多種固態或液態的介質，其對哺乳動物不具毒性且不影響一組成物中活性成分的生物活性。
本發明提供一種減少紫外線所致皮膚損傷的組成物，係由複數種植物萃取物所組成。該組成物藉由混合菠菜、紅茶、綠茶、四季春茶、紅酒、藍莓、葡萄籽、柑橘、或青咖啡豆的萃取物而製得。以下實施例中揭露該組成物能大幅抑制紫外線A照射所致皮膚纖維母細胞死亡。材料與方法 材料
含有Earle’s平衡鹽溶液之Eagle’s最低基本培養基(Eagle’s minimum essential medium，簡稱MEM培養基)，胎牛血清(fetal bovine serum，FBS)，青黴素/鏈黴素(penicillin/streptomycin)，及磷酸緩衝鹽溶液(phosphate buffered saline，PBS溶液)係自Gibco購買。用於細胞存活分析之3-(4,5-二甲基-2-噻唑)-2,5-二苯基四氮唑溴鹽(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide，MTT)係自AMERSCO購買。二甲基亞碸(dimethyl sulfoxide，DMSO)係自Echo chemical購買。蘋果多酚係(apple polyphenols)係自佶旺有限公司購買。細胞培養
以下實施例使用人類皮膚纖維母細胞(human skin fibroblast) CCD-966SK (ATCC CRL-1881)進行實驗。人類皮膚纖維母細胞在37°C、5%二氧化碳的條件下培養於添加10% FBS及1%青黴素/鏈黴素之MEM培養基，以下稱細胞培養基。MTT 分析
細胞存活率或增生率係以MTT分析測定。簡言之，將MTT溶液(4 mg/ml MTT溶於PBS溶液)依15 μl/孔添加至96孔盤中的細胞，於室溫反應4小時。移除反應液後，將DMSO依50 μl/孔添加至細胞並震盪反應10分鐘以溶解所生成的甲䐶(formazan)結晶。最終，使用ELISA讀盤機(enzyme-linked immunosorbent assay reader；BioTek)測量該細胞混合物在570 nm的吸光值(O.D. 570)。統計上顯著差異係以Excel軟體之學生t檢定進行分析。實施例 1 植物萃取物之製備 實施例 1-1 菠菜萃取物
菠菜萃取物係對菠菜(Spinacia oleracea
)進行萃取而獲得，該萃取物可購自弘祥農產加工廠。實施例 1-2 紅茶萃取物
本實施例舉例說明紅茶(black tea)萃取物之製備方法。首先，將紅茶茶葉(fermented leaves ofCamellia sinensis
)洗淨及乾燥，以粉碎機將茶葉粗碎。其次，以水為溶劑對紅茶粗碎物進行萃取，該溶劑與紅茶粗碎物依液固比5~20：1~5均勻混合，萃取溫度為介於50°C~100°C，較佳為75°C~95°C。萃取時間約為0.5~3小時。經該萃取步驟所得紅茶萃取物冷卻至室溫後，經由400目(mesh)之濾網過濾以移除殘餘固體物。該過濾後的紅茶萃取物可進一步在45°C~70°C進行減壓濃縮而獲得一濃縮產物。
在本實施例中，較佳地，水與紅茶粗碎物可依液固比5:1的比例混合後，於75℃下對紅茶粗碎物進行萃取3小時，經前述萃取步驟所得紅茶萃取物冷卻至室溫後，以400目(mesh)濾網過濾以移除殘餘固體物。過濾後的紅茶萃取物可進一步在70℃下進行減壓濃縮而獲得一濃縮產物。實施例 1-3 綠茶萃取物
本實施例舉例說明綠茶(green tea)萃取物之製備方法。首先，將綠茶茶葉(unfermented leaves ofCamellia sinensis
)洗淨及乾燥，以粉碎機將茶葉粗碎。其次，以水為溶劑對綠茶粗碎物進行萃取，該溶劑與綠茶粗碎物依液固比5~20：1~5均勻混合，萃取溫度為介於50°C~100°C，較佳為75°C~95°C。萃取時間約為0.5~3小時。經該萃取步驟所得綠茶萃取物冷卻至室溫後，經由400目之濾網過濾以移除殘餘固體物。該過濾後的綠茶萃取物可進一步在45°C~70°C進行減壓濃縮而獲得一濃縮產物。
在本實施例中，較佳地，水與綠茶粗碎物可依液固比5:1的比例混合後，於75℃下對綠茶粗碎物進行萃取3小時，經前述萃取步驟所得綠茶萃取物冷卻至室溫後，以400目(mesh)濾網過濾以移除殘餘固體物。過濾後的綠茶萃取物可進一步在70℃下進行減壓濃縮而獲得一濃縮產物。實施例 1-4 普洱茶萃取物
普洱茶(Puerh tea)萃取物係對普洱茶茶葉(post-fermented leaves ofCamellia sinensis
)進行萃取而獲得，該萃取物可購自南京澤朗生物科技有限公司。實施例 1-5 四季春茶萃取物
本實施例舉例說明四季春茶(Four Seasons Spring tea)萃取物之製備方法。首先，將四季春茶茶葉(leaves of Four Seasons Spring tea plant)洗淨及乾燥，以粉碎機將茶葉粗碎。其次，以水為溶劑對四季春茶粗碎物進行萃取，該溶劑與四季春茶粗碎物依液固比5~20：1~5均勻混合，萃取溫度為介於50°C~100°C，較佳為75°C~95°C。萃取時間約為0.5~3小時。經該萃取步驟所得四季春茶萃取物冷卻至室溫後，經由400目之濾網過濾以移除殘餘固體物。該過濾後的四季春茶萃取物可進一步在45°C~70°C進行減壓濃縮而獲得一濃縮產物。
在本實施例中，較佳地，水與四季春茶粗碎物可依液固比5:1的比例混合後，於75℃下對四季春茶粗碎物進行萃取3小時，經前述萃取步驟所得四季春茶萃取物冷卻至室溫後，以400目(mesh)濾網過濾以移除殘餘固體物。過濾後的四季春茶萃取物可進一步在70℃下進行減壓濃縮而獲得一濃縮產物。實施例 1-6 紅酒萃取物
紅酒萃取物係對紅葡萄酒進行萃取而獲得，該萃取物可購自上海博優堂生物技術有限公司。實施例 1-7 青咖啡豆萃取物
青咖啡豆萃取物係對未經烘焙的咖啡屬植物(Coffea spp.)種子進行萃取而獲得，該萃取物可購自ARJUNA NATURAL EXTRACTS有限公司(印度)。實施例 1-8 藍莓萃取物
藍莓萃取物係對北美藍莓(Vaccinium Cyanococcus
)果實進行萃取而獲得，該萃取物可購自元弘草本科研有限公司。實施例 1-9 柑橘萃取物
柑橘萃取物係對柑橘(Citrus reticulata
)果實進行萃取而獲得，該萃取物可購自羅頓貿易有限公司。實施例 1-10 葡萄籽萃取物
葡萄籽萃取物係對葡萄屬(Vitis spp.)植物種子進行萃取而獲得，該萃取物可購自誠鴻生物科技有限公司。實施例 1-11 迷迭香萃取 物
迷迭香萃取物係對迷迭香(Rosmarinus officinalis
)進行萃取而獲得，該萃取物可購自佳景百草股份有限公司。實施例 2 含植物萃取物之組成物抑制紫外線 A 照射所致皮膚纖維母細胞死亡
為檢驗本發明之組成物保護皮膚對抗紫外線照射的作用，利用細胞存活分析(MTT分析)評估經過紫外線A照射的人類皮膚纖維母細胞CCD-966SK在植物萃取物或其組合處理後的細胞增生率。簡言之，將CCD-966SK細胞依5×103
個細胞/孔接種於含有200 μl細胞培養基的96孔培養盤，在37°C下培養24小時後，移除細胞培養基，並使細胞在紫外線照射箱(Vilber)中接受15 J/cm2
紫外線A照射1小時，此輻射劑量會造成半數細胞死亡。其後，將表1所列植物萃取物或含有植物萃取物的組成物連同細胞培養基添加至細胞，於37°C下培養24小時。另設置一組經紫外線A照射但以不含植物萃取物之細胞培養基處理的細胞，其為負控制組；以及一組未予紫外線A照射且僅以不含植物萃取物之細胞培養基處理的細胞，其為空白對照組。最終，進行MTT分析及計算各組細胞之細胞增生率。該細胞增生率依下列公式計算：
細胞增生率 = 各組的O.D. 570/ 負控制組的O.D. 570×100%
表1
| 處理 | 細胞增生率 | |
| 空白對照組 | 237% | |
| 負控制組 | 100% | |
| 菠菜 | 0.25 mg/ml | 138% |
| 0.5 mg/ml | 155% | |
| 柑橘 | 0.5 mg/m | 142% |
| 紅茶 | 0.03125 mg/ml | 96.39% |
| 綠茶 | 0.25 mg/ml | 139.36% |
| 0.0625 mg/ml | 110.04% | |
| 普洱茶 | 0.03125 mg/ml | 96.79% |
| 四季春茶 | 0.25 mg/ml | 104.02% |
| 0.125 mg/ml | 100.40% | |
| 青咖啡豆 | 0.015625 mg/ml | 127.31% |
| 紅酒 | 0.25 mg/ml | 169.88% |
| 藍莓 | 0.5 mg/ml | 109.24% |
| 葡萄籽 | 0.078125 mg/ml | 123.69% |
| 菠菜+紅茶 0.25 mg/ml+0.25 mg/ml | 180% | |
| 菠菜+綠茶 0.25 mg/ml+0.25 mg/ml | 140% | |
| 菠菜+普洱茶 0.25 mg/ml+0.25 mg/ml | 110% | |
| 菠菜+四季春茶 0.25 mg/ml+0.25 mg/ml | 140% | |
| 菠菜+青咖啡豆 0.125 mg/ml+0.125 mg/ml | 60% | |
| 菠菜+紅酒 0.25 mg/ml+0.25 mg/ml | 250% | |
| 菠菜+藍莓 0.5 mg/ml+0.5 mg/ml | 220% | |
| 菠菜+葡萄籽 0.078125 mg/ml+0.078125 mg/ml | 168% | |
| 菠菜+蘋果多酚 0.0625 mg/ml+0.0625 mg/ml | 126% | |
| 菠菜+迷迭香 0.125 mg/ml+0.125 mg/ml | 115% | |
| 柑橘+紅茶 0.0625 mg/ml+0.0625 mg/ml | 142% | |
| 柑橘+綠茶 0.0625 mg/ml+0.0625 mg/ml | 176% | |
| 柑橘+普洱茶 0.03125 mg/ml+0.03125 mg/ml | 193% | |
| 柑橘+四季春茶 0.125 mg/ml+0.125 mg/ml | 143% | |
| 柑橘+菠菜 0.25 mg/ml+0.25 mg/ml | 204% | |
| 柑橘+青咖啡豆 0.015625 mg/ml+0.015625 mg/ml | 167% | |
| 柑橘+紅酒 0.25 mg/ml+0.25 mg/ml | 209% | |
| 柑橘+藍莓 0.5 mg/ml+0.5 mg/ml | 93% | |
| 柑橘+葡萄籽 0.078125 mg/ml+0.078125 mg/ml | 166% | |
| 柑橘+蘋果多酚 0.0625 mg/ml+0.0625 mg/ml | 114% |
表1顯示皮膚纖維母細胞在不同處理後的細胞增生率；圖1是對應表1所列數值的長條圖。依據表1，空白對照組相對於負控制組的細胞增生率為237%，顯示紫外線A照射會造成皮膚纖維母細胞大量死亡。依據表1及圖1，相對於負控制組，菠菜萃取物(0.25 mg/ml)或柑橘萃取物(0.5 mg/ml)之單獨處理使細胞增生率分別回升至138%及142%。此外，紅茶萃取物、綠茶萃取物、普洱茶萃取物、四季春茶萃取物、青咖啡豆萃取物、藍莓萃取物、或葡萄籽萃取物之單獨處理使細胞增生率在約95%至140%之間變動。紅酒萃取物之單獨處理使細胞增生率約為170%。
值得注意的是菠菜萃取物與紅茶萃取物、藍莓萃取物、或葡萄籽萃取物的組合提高皮膚纖維母細胞的細胞增生率至明顯高於100%至138%；菠菜萃取物與紅酒萃取物的組合提高細胞增生率至約250%。同樣地，濃度等於或遠低於0.5 mg/ml之柑橘萃取物與紅茶萃取物、綠茶萃取物、普洱茶萃取物、四季春茶萃取物、青咖啡豆萃取物、菠菜萃取物、或葡萄籽萃取物的組合會提高細胞增生率至明顯高於100%至142%；柑橘萃取物與紅酒萃取物的組合提高細胞增生率至約209%。具有上列特定組合的組成物皆意外地較單一組分簡單相加表現出對抗紫外線A的更高保護力。
綜上所述，本發明之組成物藉由混合特定植物萃取物，抑制紫外線照射所致皮膚纖維母細胞死亡。皮膚纖維母細胞的正常增生則有利於維持表皮的厚度及紫外線屏障功能，亦可補充皮膚細胞外基質中的組成及維持真皮的更新。故，本發明之組成物可連同藥學上可接受之載劑用於製備減少紫外線所致皮膚損傷的醫藥組成物。該醫藥組成物之劑型，可為但不限於溶液、粉末、膠囊、或錠劑。
無。
圖1顯示本發明一實施例中不同組成物對紫外線照射下皮膚纖維母細胞之細胞增生率的影響。
無。
Claims (7)
- 一種修護受損皮膚纖維母細胞的組成物，其中該組成物係選自柑橘萃取物與紅茶萃取物、普洱茶萃取物、四季春茶萃取物、紅酒萃取物、及葡萄籽萃取物其中任一種，其中該組成物中各該萃取物之比例為1：1。
- 如申請專利範圍第1項所述之組成物，係由下列任一組合所組成：至少0.03125 mg/ml之柑橘萃取物，及至少0.03125 mg/ml之普洱茶萃取物；至少0.0625 mg/ml之柑橘萃取物，及至少0.0625 mg/ml之紅茶萃取物；至少0.078125 mg/ml之柑橘萃取物，及至少0.078125 mg/ml之葡萄籽萃取物；至少0.125 mg/ml之柑橘萃取物，及至少0.125 mg/ml之四季春茶萃取物；或至少0.25 mg/ml之柑橘萃取物，及至少0.25 mg/ml之紅酒萃取物。
- 一種用於製備修護受損皮膚纖維母細胞之醫藥組成物的用途，該醫藥組成物包含如申請專利範圍第1項或第2項所述之組成物，及一藥學上可接受之載劑。
- 如申請專利範圍第3項所述之用途，其中該醫藥組成物具有溶液、粉末、膠囊、或錠劑之劑型。
- 如申請專利範圍第3項所述之用途，其中該醫藥組成物可以提升皮膚纖維母細胞增生率。
- 如申請專利範圍第3項所述之用途，其中該醫藥組成物可以補充皮膚細胞外基質中的組成及維持真皮的更新效率。
- 如申請專利範圍第3項所述之用途，其中該醫藥組成物可以維持表皮的厚度及提升表皮的紫外線屏障功能。
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| CN105639355A (zh) * | 2016-03-30 | 2016-06-08 | 天地壹号饮料股份有限公司 | 一种苹果果胶膳食纤维饮料及其制备方法 |
| CN105851316A (zh) * | 2016-04-21 | 2016-08-17 | 华南农业大学 | 柑橘乌龙茶及其制备方法 |
| CN105995696A (zh) * | 2016-05-20 | 2016-10-12 | 安徽省华之慧生态农业发展有限公司 | 一种富含膳食纤维的益生元夹心蔬菜纸及其制备方法 |
| CN105941728A (zh) | 2016-05-24 | 2016-09-21 | 安徽源和堂药业股份有限公司 | 一种橘子皮泡腾茶饮颗粒 |
| CN105875963A (zh) | 2016-06-02 | 2016-08-24 | 江苏丰年果品种植园有限公司 | 一种蓝莓速溶茶的制备方法 |
| CN106085749A (zh) * | 2016-06-18 | 2016-11-09 | 吴金珠 | 一种带有减肥功能的保健饮料 |
| CN106075179A (zh) * | 2016-06-20 | 2016-11-09 | 威海百合生物技术股份有限公司 | 一种减肥软胶囊 |
| CN105963208A (zh) * | 2016-06-29 | 2016-09-28 | 林序彬 | 一种植物多功效洗面奶 |
| CN106165869A (zh) * | 2016-07-02 | 2016-11-30 | 马玉玲 | 一种改善记忆功能的保健食品及其制备方法 |
| CN106070466A (zh) * | 2016-07-28 | 2016-11-09 | 合肥吴复和食品有限公司 | 一种消脂瘦身山药陈皮糕 |
| CN106265415A (zh) | 2016-08-11 | 2017-01-04 | 胡安平 | 一种具有保湿和晒后修护功能的护肤组合物及其应用 |
| CN106136066A (zh) | 2016-08-25 | 2016-11-23 | 普安县宏鑫茶业开发有限公司 | 一种绿茶面条及其制作方法 |
| CN106418546A (zh) | 2016-09-22 | 2017-02-22 | 上海承煌实业有限公司 | 一种蓝莓叶黄素 |
| KR20160126932A (ko) * | 2016-10-12 | 2016-11-02 | 우상호 | 기능성 워터리스 세정제 조성물 및 이를 포함하는 워터리스 세정제 |
| CN106492084A (zh) * | 2016-11-16 | 2017-03-15 | 北京澳特舒尔保健品开发有限公司 | 一种纯草本中药减肥产品及其制备方法 |
| CN106509235A (zh) * | 2016-12-06 | 2017-03-22 | 广西职业技术学院 | 蜜香橘红茶的制作方法 |
| TWI671081B (zh) * | 2017-04-03 | 2019-09-11 | 大江生醫股份有限公司 | 用以提升轉谷氨醯胺酶１基因、角蛋白基因、水通道蛋白３基因、聚角蛋白微絲基因、葡萄糖腦苷酯酶基因、及玻尿酸合成酶基因表現之含有植物萃取物之組成物的用途 |
| CN109706018A (zh) * | 2017-10-23 | 2019-05-03 | 谢玮 | 一种红酒泡腾片及其制备方法 |
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