TW202110468A - 薔薇屬植物萃取物用於抗老化及抗氧化之用途 - Google Patents
薔薇屬植物萃取物用於抗老化及抗氧化之用途 Download PDFInfo
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Abstract
本發明提供一種薔薇屬植物萃取物用於抗老化及抗氧化之用途。
Description
本發明是有關於一種薔薇屬植物萃取物用於抗老化及抗氧化之用途。
皮膚是保護人類個體的最大屏障,它具有對抗水分散失、病原菌以及各種環境損害之功能。暴露於大量的3C藍光(例如手機及平板電腦)、紫外線(ultraviolet,UV)、游離輻射(ionizing radiation)、藥物或異生物質(xenobiotics)會促使皮膚生成活性氧族(reactive oxygen species,ROS)以及自由基(free radicals)。當所累積的活性氧族以及自由基的數量超過細胞或組織本身的抗氧化能力時,便會形成氧化性壓力(oxidative stress)。接著,活性氧族以及自由基會與細胞內的組成物(包括DNA、蛋白質以及脂質等)相反應,進而對皮膚產生非所欲的影響。
黑色素生成(melanogenesis)(亦即黑色素合成(melanin synthesis))是指當皮膚黑色素細胞(dermal melanocyte)在受到環境因素(諸如藍光、紫外線(ultraviolet,UV))或生理因素(諸如疲勞(fatigue)、壓力(stress)、慢性發炎(chronic inflammation)以及體內不正常的α-促黑素細胞素(α-melanocyte stimulating hormone,α-MSH)的釋放)的誘發之後,在黑色素細胞內的酪胺酸(tyrosine)經由酪胺酸酶(tyrosinase)的催化(它是黑色素生成的速率-限制步驟(rate-limiting step))以及一系列的氧化還原反應而被轉化為黑色素(melanin)的過程。黑色素可以保護皮膚的下皮層(hypodermis)免於紫外線所造成的光損害(photodamage),但是當黑色素被大量地累積於皮膚上或不正常地分佈時可能會導致皮膚疾病(skin
disorders),諸如雀斑(lentigines)、斑點(freckle)、黑皮病(melasma)、老人斑(age spots)以及色素過多(hyperpigmentation)等。
近年來,人類對於抑制黑色素生成及減少細胞的自由基生成的需求與日俱增,因為一旦抑制黑色素生成及減少細胞的自由基生成,就能夠達到抗老化及抗氧化的效用。然而,目前常見用來抑制黑色素生成及減少細胞的自由基生成的方式大多為利用塗抹於皮膚表面的化妝品、保養品,或口服宣稱具有抑制黑色素生成及減少細胞的自由基生成功效的健康食品。然而,習知的化妝品、保養品及健康食品大多由化學成分所製成,長期使用不但對人體健康有害無益,且這些產品往往價格昂貴,並非為一般使用者所能負擔。
另一方面,粒線體(mitochondria)亦被稱為細胞的發電站,因為它是細胞內合成三磷酸腺苷(adenosine triphosphate,ATP)(一種傳遞能量的分子)的主要場所,為細胞的各項活動提供了化學能量。粒線體若損壞,對細胞以及生物個體的影響甚鉅。粒線體在合成ATP的過程中會產生很多的自由基,自由基的活性極強,會與體內任何物質發生強烈的氧化反應而破壞其正常功能。自由基日積月累地傷害粒線體內的酵素與DNA,漸漸地使其功能下降進而使各器官組織的功能衰退。因此,如何提升細胞的粒線體活性,進而達到抗老化及抗氧化之效用,成為本領域的重要課題。
為了解決上述問題,本領域的技術人員亟需研發出具有提升細胞的粒線體活性、減少細胞的自由基生成、抑制黑色素生成、抗老化及抗氧化效用的新穎醫藥品、食品產品或保養品以造福有此需求的廣大族群。
有鑑於此,本發明之目的為提供一種薔薇屬(ROSA)植物萃取物用於製備一抗老化及抗氧化之組成物的用途,其中該薔薇屬植物萃取物係以一溶劑萃取一薔薇屬植物所獲得,該溶劑為水、甘油、含水甘油或其組合,其中該薔薇屬植物萃取物是一玫瑰(Rosa rugosa)花萃取物。
在本發明的一實施例中,該抗老化及抗氧化包括提升細胞的粒線體活性、減少細胞的自由基生成及抑制黑色素生成。
在本發明的一實施例中,該細胞是一皮膚纖維母細胞。
在本發明的一實施例中,該自由基生成是由一藍光或一過氧化氫所導致。
在本發明的一實施例中,該黑色素生成是由一藍光所導致。
在本發明的一實施例中,該玫瑰花萃取物的有效濃度為至少0.25%(v/v)。
在本發明的一實施例中,該萃取是於一介於65~85℃的溫度進行。
在本發明的一實施例中,該溶劑與該玫瑰花的體積比介於5~20:1~5。
在本發明的一實施例中,該組成物是一醫藥品、一食品產品或一保養品。
在本發明的一實施例中,該醫藥品包含一醫藥上可接受的載劑。
綜上所述,本發明薔薇屬植物萃取物之功效在於:可藉由提升細胞(例如皮膚纖維母細胞)的粒線體活性、減少細胞(例如皮膚纖維母細胞)的自由基生成(例如由藍光所導致)及抑制黑色素生成(例如由3C藍光產品如手機及平板電腦所導致),達到保護皮膚細胞,強力抗氧化及抗老化的功效,藉此維持肌膚年輕活力。
以下將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,並非用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。
圖1是本發明薔薇屬植物萃取物萃取物在提升皮膚纖維母細胞的粒線體活性上之效用的數據圖,其中“***”表示與對照組比較,p<0.001。
圖2是本發明薔薇屬植物萃取物在減少由藍光所導致的細胞自由基生成上的功效之數據圖,其中“**”表示p<0.01。
圖3是本發明薔薇屬植物萃取物在減少由過氧化氫所導致的細胞自由基生成上的功效之數據圖。
圖4是本發明薔薇屬植物萃取物在抑制藍光所導致的黑色素生成上的功效之數據圖。
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。
使用Excel軟體進行統計分析。數據以平均值±標準差(standard deviation,STDEV)表示,個此之間的差異以學生t檢驗(student's t-test)分析。
本發明使用的植物材料玫瑰(Rosa rugosa)是一種薔薇科(Rosaceae)薔薇屬(Rosa)的植物。明代田汝成《西湖游覽志馀.委巷叢談》提到『玫瑰花雜腦麝以為香囊,芬氤裊裊不絕』,故又名徘徊花,稱之其香氣悠長繚繞,受人喜愛。
如本文中所使用的,用語「抗老化(anti-aging)」意指預防、減緩人類皮膚外觀之老化現象,例如:皺紋的產生及失去彈性等。評量實現此目的之程度將根據熟悉此項技藝者已知之諸多因素來決定,諸如消費者的全身狀態、年齡、性別等。
如本文中所使用的,用語「抑制黑色素生成(inhibition of melanogenesis)」與「抑制黑色素合成(inhibition of melanin synthesis)」、「去色素(depigmenting)」、「淡化黑色素(lightening the melanin)」、「美白(whitening)」、「膚色淡化(skin color lightening)」、「漂白(bleaching)」、「淨白」、「增白(brightening)」、「退黑」以及「驅黑」可被交換地使用。
依據本發明,醫藥品可利用熟習此技藝者所詳知的技術而被製造成一適合於非經腸道地(parenterally)、口服地(orally)或局部地(topically)投藥
的劑型,這包括,但不限於:注射品(injection)[例如,無菌的水性溶液(sterile aqueous solution)或分散液(dispersion)]、無菌的粉末(sterile powder)、錠劑(tablet)、片劑(troche)、口含錠(lozenge)、丸劑(pill)、膠囊(capsule)、分散性粉末(dispersible powder)或細顆粒(granule)、溶液、懸浮液(suspension)、乳劑(emulsion)、糖漿(syrup)、酏劑(elixir)、濃漿(slurry)、外部製劑(external preparation)以及類似之物。
依據本發明,醫藥品可進一步包含有一被廣泛地使用於藥物製造技術之醫藥上可接受的載劑(pharmaceutically acceptable carrier)。例如,該醫藥上可接受的載劑可包含一或多種選自於下列的試劑:溶劑(solvent)、緩衝液(buffer)、乳化劑(emulsifier)、懸浮劑(suspending agent)、分解劑(decomposer)、崩解劑(disintegrating agent)、分散劑(dispersing agent)、黏結劑(binding agent)、賦形劑(excipient)、安定劑(stabilizing agent)、螯合劑(chelating agent)、稀釋劑(diluent)、膠凝劑(gelling agent)、防腐劑(preservative)、潤濕劑(wetting agent)、潤滑劑(lubricant)、吸收延遲劑(absorption delaying agent)、脂質體(liposome)以及類似之物。有關這些試劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。
依據本發明,該醫藥上可接受的載劑包含有一選自於由下列所構成之群組中的溶劑:水、生理鹽水(normal saline)、磷酸鹽緩衝生理鹽水(phosphate buffered saline,PBS)、含有醇的水性溶液(aqueous solution containing alcohol)以及它們的組合。
依據本發明,該醫藥品可以一選自於由下列所構成之群組中的非經腸道途徑(parenteral routes)來投藥:腹膜內注射(intraperitoneal injection)、皮下注射(subcutaneous injection)、表皮內注射(intraepidermal injection)、皮內注射(intradermal injection)、肌肉內注射(intramuscular injection)、靜脈內注射(intravenous injection)以及病灶內注射(intralesional injection)。
依據本發明,醫藥品可利用熟習此技藝者所詳知的技術而被製造成一適合於局部地施用於皮膚上的外部製劑(external preparation),這包括,但不限於:乳劑(emulsion)、凝膠(gel)、軟膏(ointment)、乳霜(cream)、貼片
(patch)、擦劑(liniment)、粉末(powder)、氣溶膠(aerosol)、噴霧(spray)、乳液(lotion)、乳漿(serum)、糊劑(paste)、泡沫(foam)、滴劑(drop)、懸浮液(suspension)、油膏(salve)以及繃帶(bandage)。
依據本發明,該外部製劑是藉由將本發明的醫藥品與一為熟習此項技藝者所詳知的基底(base)相混合而被製備。
依據本發明,該基底可包含有一或多種選自於下列的添加劑(additives):水、醇(alcohols)、甘醇(glycol)、碳氫化合物(hydrocarbons)[諸如石油膠(petroleum,jelly)以及白凡士林(white petrolatum)]、蠟(wax)[諸如石蠟(paraffin)以及黃蠟(yellow wax)]、保存劑(preserving agents)、抗氧化劑(antioxidants)、界面活性劑(surfactants)、吸收增強劑(absorption enhancers)、安定劑(stabilizing agents)、膠凝劑(gelling agents)[諸如卡波普®974P(carbopol®974P)、微結晶纖維素(microcrystalline cellulose)以及羧基甲基纖維素(carboxymethylcellulose)]、活性劑(active agents)、保濕劑(humectants)、氣味吸收劑(odor absorbers)、香料(fragrances)、pH調整劑(pH adjusting agents)、螯合劑(chelating agents)、乳化劑(emulsifiers)、閉塞劑(occlusive agents)、軟化劑(emollients)、增稠劑(thickeners)、助溶劑(solubilizing agents)、滲透增強劑(penetration enhancers)、抗刺激劑(anti-irritants)、著色劑(colorants)以及推進劑(propellants)等。有關這些添加劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。
依據本發明,保養品可進一步包含有一被廣泛地使用於保養品製造技術之可接受的佐劑(acceptable adjuvant)。例如,該可接受的佐劑可包含有一或多種選自於下列的試劑:溶劑、膠凝劑、活性劑、防腐劑、抗氧化劑、遮蔽劑(screening agent)、螯合劑、界面活性劑、染色試劑(coloring agent)、增稠劑(thickening agent)、填料(filler)、香料以及氣味吸收劑。有關這些試劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。
依據本發明,保養品可利用熟習此技藝者所詳知的技術而被製造成一適合於護膚(skincare)或化妝(makeup)的形式,這包括,但不限於:水性溶液(aqueous solution)、水-醇溶液(aqueous-alcohol solution)或油性溶液(oily
solution)、呈水包油型(oil-in-water type)、油包水型(water-in-oil type)或複合型之乳劑、凝膠、軟膏、乳霜、面膜(mask)、貼片、貼布(pack)、擦劑、粉末、氣溶膠、噴霧、乳液、乳漿、糊劑、泡沫、分散液、滴劑、慕斯(mousse)、防曬油(sunblock)、化妝水(tonic water)、粉底(foundation)、卸妝產品(makeup remover products)、肥皂(soap)以及其他身體清潔產品(body cleansing products)等。
依據本發明,保養品亦可與一或多種選自於下列之已知活性的外用劑(external use agents)一起合併使用:美白劑(whitening agents)[諸如維生素A酸(tretinoin)、兒茶素(catechin)、麴酸、熊果苷以及維生素C]、保濕劑、抗發炎劑(anti-inflammatory agents)、殺菌劑(bactericides)、紫外線吸收劑(ultraviolet absorbers)、植物萃取物(plant extracts)[諸如蘆薈萃取物(aloe extract)]、皮膚營養劑(skin nutrients)、麻醉劑(anesthetics)、抗痘劑(anti-acne agents)、止癢劑(antipruritics)、止痛劑(analgesics)、抗皮膚炎劑(antidermatitis agents)、抗過角化劑(antihyperkeratolytic agents)、抗乾皮膚劑(anti-dry skin agents)、抗汗劑(antipsoriatic agents)、抗老化劑(antiaging agents)、抗皺劑(antiwrinkle agents)、抗皮脂溢出劑(antiseborrheic agents)、傷口治療劑(wound-healing agents)、皮質類固醇(corticosteroids)以及激素(hormones)。有關這些外用劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。
依據本發明,食品產品可被當作食品添加物(food additive),藉由習知方法於原料製備時添加,或是於食品的製作過程中添加,而與任一種可食性材料配製成供人類與非人類動物攝食的食品產品。
依據本發明,食品產品的種類包括但不限於:飲料(beverages)、發酵食品(fermented foods)、烘培產品(bakery products)、健康食品(health foods)以及膳食補充品(dietary supplements)。
首先,對薔薇屬(ROSA)植物(較佳為玫瑰花(Rosa rugosa))(購自於宣洋實業股份有限公司的伊朗產的玫瑰乾料)進行均質處理,然後於65~85℃下,以溶劑對經均質處理的薔薇屬植物(以5~20:1~5的體積比進行萃取而得到一粗萃取物,其中溶劑是水、甘油、含水甘油或其組合。接著,以濾網對粗萃
取物過濾而得到一濾液,然後進行離心,接而收集上清液並進行過濾,而得到薔薇屬植物萃取物。
本實施例以人類皮膚纖維母細胞CCD-966SK進行皮膚細胞粒線體活性分析,並利用流式細胞儀粒線體膜電位偵測套組(Flow cytometry Mitochrondrial membrane potential detection kit,BD)進行實驗。人類皮膚纖維母細胞購自台灣生物資源保存及研究中心(Bioresource Collection and Research Center,BCRC),編號BCRC 60153。將該細胞培養於添加10%胎牛血清(fetal bovine serum,FBS)(GIBCO公司,編號10438-026,美國)、0.1mM非必需胺基酸、1.5g/L碳酸氫鈉(Sigma公司,編號S5761,美國)、1mM丙酮酸鈉(GIBCO公司,編號11360-070,美國)的最低必需培養液(Minimum essential medium,MEM)(Eagle)(配於厄爾平衡鹽溶液(Earle's Balanced Salt Solution,Earle's BSS))(GIBCO公司,編號41500-034,美國)。
在含有2mL上述培養基的6孔培養盤中每個孔洞接種1×105個人類皮膚纖維母細胞(n=3)。之後,將細胞分成二組,其中包括對照組及實驗組。將0.5%(v/v)薔薇屬植物萃取物添加至實驗組的細胞中,至於對照組的細胞則添加培養基。接著,將各組細胞於37℃下培養24小時。在使用BDTM MitoScreen(JC-1)試劑套組前,需先將10x JC-1染色測定緩衝液(JC-1 10x assay buffer)置於37℃下預熱,並以無菌之1x磷酸鹽緩衝溶液(1x Phosphate buffered saline,1x PBS)配製1x JC-1染色測定緩衝液,且維持溫度為37℃,接著加入130μL之二甲基亞碸(Dimethyl sulfoxide,DMSO)於該1x JC-1染色測定緩衝液中進行冷凍乾燥,即為JC-1染色測定緩衝液的儲存溶液,其能夠在-20℃下保存6個月;待使用時,將JC-1染劑與該1x JC-1測定緩衝液以1:100的比例均勻,即為JC-1作用染劑。之後,加入胰蛋白酶(trypsin)/EDTA處理3分鐘後,吸取懸浮的細胞至1.5mL的微離心管,以400g轉速離心5分鐘收集沈澱的細胞。
在移除上清液後,以1mL的1X PBS再懸浮細胞,然後轉移至1.5mL的離心管,以400g轉速離心5分鐘。在移除上清液後,添加100μL的JC-1工作溶液,混合均勻後在避光下作用15分鐘。之後,以400g轉速離心5分鐘,再以
1mL的1X清洗緩衝液清洗並以400g轉速離心5分鐘,然後以1mL的1X清洗緩衝液清洗並以400g轉速離心5分鐘。以含有2%FBS的500μL之1X PBS再懸浮細胞,然後利用流式細胞儀(Beckman)分析觀察細胞凋亡時粒線體膜電位改變,並以Excel進行t-檢驗(student t-test)統計分析樣品群體之間差異的統計學意義。
圖1是本發明薔薇屬植物萃取物在提升皮膚纖維母細胞的粒線體活性上之效用的數據圖。由圖1可見,與對照組相較之下,實驗組的皮膚纖維母細胞的粒線體活性(即相對的JC-1聚合體(aggregate))有顯著提升,其中與對照組比較,實驗組的皮膚纖維母細胞的粒線體活性提升達55.4%。本實施例的結果顯示,本發明薔薇屬植物萃取物可以顯著提升細胞粒線體活性,維持肌膚年輕活力,達到抗老之潛力。
首先,以添加有10%胎牛血清(Gibco)、0.5mM丙酮酸鈉(sodium pyruvate)及15mM HEPES的杜貝可氏改良的依格氏培養基(Dulbecco’s Modified Eagle’s Medium,DMEM)與Ham's F12培養基(1:1混合)培養人類皮膚纖維母細胞CCD-966SK(BCRC 60153)於6-孔盤,2mL培養基的細胞濃度為1.5×105細胞/孔,接而於37℃進行培養24小時,並移除培養基。接著,將經培養的細胞分成三組,其中包括對照組、藍光組,及實驗組。將0.5%(v/v)薔薇屬植物萃取物添加至實驗組的細胞中,並照射藍光(品牌:ZAMI STUDIO,藍光波長:415nm;劑量50J/cm2)15分鐘。藍光組的細胞被照射藍光15分鐘,至於對照組的細胞則未添加薔薇屬植物萃取物及未照射藍光。將各組細胞於37℃進行培養1小時之後,添加5μg/mL二氯二氫螢光黃二乙酸酯(Dichloro-dihydro-fluorescein diacetate,DCFH-DA)(Sigma/SI-D6883-50MG)(儲備溶液為5mg/mL溶於DMSO中)並於37℃反應15分鐘。接著,以1mL的1X PBS(Gibco)清洗每孔兩次,然後添加200μL胰蛋白酶(trypsin)並於避光環境反應5分鐘,接而將細胞培養物收集至1.5mL體積的離心管,並以400×g進行離心10分鐘。之後,移除上清液並以1X PBS清洗一次,接而以400×g進行離心10分鐘。接著,移除上清液並以1mL的1X PBS再懸浮細胞沉澱物。之後,以流式細胞儀(Beckman)藉由激發波長(excitation wavelength)450~490nm及放射波長(emission wavelength)510~550nm
來偵測DCFH-DA的螢光訊號,藉此計算出受到ROS傷害的細胞百分比。各組之間的統計學顯著差異是藉由史徒登氏t-檢定來決定。本實施例的結果顯示於圖2。
圖2是本發明薔薇屬植物萃取物在減少由藍光所導致的細胞自由基生成上的功效之數據圖。由圖2可見,與對照組相較之下,藍光組測得的受到ROS傷害的細胞百分比(即氧化壓力高表現細胞)有顯著提升,這表示藍光會對皮膚纖維母細胞產生大量ROS傷害;而與藍光組相較之下,實驗組測得的受到ROS傷害的細胞百分比有顯著降低(降低藍光所導致的ROS表現量約21%)。本實施例的結果顯示,本發明薔薇屬植物萃取物具有減少由藍光所導致的細胞自由基生成之功效。
首先,以添加有10%胎牛血清(FBS)(GIBCO)、0.1mM非必需胺基酸、1.5g/L碳酸氫鈉及1mM丙酮酸鈉(sodium pyruvate)(90%)的最低必需培養基(MEM)(Eagle)(配於厄爾平衡鹽溶液(Earle's Balanced Salt Solution,Earle's BSS))(GIBCO公司,編號41500-034,美國)培養人類皮膚纖維母細胞CCD-966SK(BCRC 60153)於6-孔盤,2mL培養基的細胞濃度為2×105細胞/孔,接而於37℃進行培養24小時,並移除培養基。接著,將經培養的細胞分成三組,其中包括對照組、過氧化氫組,及實驗組。將0.25%(v/v)薔薇屬植物萃取物及1mM H2O2(Sigma)添加至實驗組的細胞中。過氧化氫組的細胞被添加以1mM H2O2,至於對照組的細胞則不做任何處理。之後,添加5μg/mL二氯二氫螢光黃二乙酸酯(Dichloro-dihydro-fluorescein diacetate,DCFH-DA)(Sigma/SI-D6883-50MG)(儲備溶液為5mg/mL溶於DMSO中)並於37℃反應15分鐘,然後於37℃下以H2O2處理細胞1小時,接而以1mL的1X PBS(Gibco)清洗每孔兩次。接著,添加200μL胰蛋白酶(trypsin)並於避光環境反應5分鐘,接而將細胞培養物收集至1.5mL體積的離心管,並以400×g進行離心10分鐘。之後,移除上清液並以1X PBS清洗一次,接而以400×g進行離心10分鐘。接著,移除上清液並以1mL的1X PBS再懸浮細胞沉澱物。之後,以流式細胞儀(Beckman)藉由激發波長(excitation wavelength)450~490nm及放射波長(emission wavelength)510~550nm來偵測
DCFH-DA的螢光訊號,藉此計算出受到ROS傷害的細胞百分比。各組之間的統計學顯著差異是藉由史徒登氏t-檢定來決定。本實施例的結果顯示於圖3。
圖3是本發明薔薇屬植物萃取物在減少由過氧化氫所導致的細胞自由基生成上的功效之數據圖。由圖3可見,與對照組相較之下,過氧化氫組測得的受到ROS傷害的細胞百分比(即氧化壓力高表現細胞)有顯著提升,這表示過氧化氫會對皮膚纖維母細胞產生大量ROS傷害;而與過氧化氫組相較之下,實驗組測得的受到ROS傷害的細胞百分比有顯著降低(降低ROS表現量至少97%)。本實施例的結果顯示,本發明薔薇屬植物萃取物具有減少由過氧化氫所導致的細胞自由基生成的功效。
首先,將老鼠皮膚黑色素瘤細胞株B16F10(對應於ATCC CRL-6475)培養於杜貝可氏改良的依格氏培養基(Dulbecco’s Modified Eagle’s Medium,DMEM)(添加有1%青黴素/鏈黴素(Gibco)及10% FBS(Gibco))中。於6孔培養盤的每孔中加入3mL的培養基,使每孔具有1.5 x 105個B16F10細胞。在37℃下培養24小時後,移除培養基。
之後,將B16F10細胞分成三組,其中包括實驗組、藍光組以及對照組。將0.5%(v/v)薔薇屬植物萃取物添加至實驗組的細胞中,於37℃下培養1小時後,並照射藍光3小時。藍光組的細胞被照射藍光3小時,至於對照組的細胞則未添加薔薇屬植物萃取物及未照射藍光。
各組細胞培養物在37℃下培養48小時後,移除培養基並以1xPBS(Gibco)沖洗兩次。之後,加入胰蛋白酶(trypsin)來處理細胞3分鐘並將懸浮的細胞收集於15mL體積的離心管中,接而以400xg/5分鐘進行旋轉(spin)以沉澱細胞。在以1xPBS沖洗兩次之後,以200μL的1XPBS再懸浮細胞沉澱物(cell pellet)。接著,將細胞溶液於液態氮中放置10分鐘,接而於室溫下靜置30分鐘進行解凍。在解凍完全之後,以12,000xg進行旋轉30分鐘,接而移除上清液並添加120μL的1N NaOH(配於ddH2O)。在混合均勻之後,於60℃乾浴槽中靜置1小時。之後,取100μL的體積至96孔培養盤中並於450nm的波長下以ELISA讀取儀來讀取各孔的吸光值(OD450)。
黑色素含量(%)是藉由將所測得的吸光值(OD450)代入下列公式(1)而計算出:
黑色素含量(%)=(各組所測得的OD450吸光值/對照組所測得的OD450吸光值)×100% (1)
各組之間的統計學顯著差異是藉由史徒登氏t-檢定來決定。本實施例的結果顯示於圖4。
圖4是本發明薔薇屬植物萃取物在抑制藍光所導致的黑色素生成上的功效之數據圖。由圖4可見,與對照組相較之下,藍光組的黑色素含量有顯著提升,這表示藍光會對細胞產生大量黑色素;而與藍光組相較之下,實驗組的黑色素含量有顯著的降低(與藍光組比較,實驗組的黑色素含量降低至少38.8%)。本實施例的結果顯示,本發明薔薇屬植物花萃取物具有抑制藍光所導致的黑色素生成的功效。
綜上所述,本發明薔薇屬植物萃取物可藉由提升細胞(例如皮膚纖維母細胞)的粒線體活性、減少細胞(例如皮膚纖維母細胞)的自由基生成(例如由藍光所導致)及抑制黑色素生成(例如由3C藍光產品如手機及平板電腦所導致),達到保護皮膚細胞,強力抗氧化及抗老化的功效,藉此維持肌膚年輕活力。
以上所述僅為舉例性,而非為限制性者。任何未脫離本發明之精神與範疇,而對其進行之等效修改或變更,均應包含於後附之申請專利範圍中。
Claims (10)
- 一種薔薇屬(ROSA)植物萃取物用於製備一抗老化及抗氧化之組成物的用途,其中該薔薇屬植物萃取物係以一溶劑萃取一薔薇屬植物所獲得,該溶劑為水、甘油、含水甘油或其組合,其中該薔薇屬植物萃取物是一玫瑰(Rosa rugosa)花萃取物。
- 如申請專利範圍第1項所述的用途,其中該抗老化及抗氧化包括提升細胞的粒線體活性、減少細胞的自由基生成及抑制黑色素生成。
- 如申請專利範圍第2項所述的用途,其中該細胞是一皮膚纖維母細胞。
- 如申請專利範圍第3項所述的用途,其中該自由基生成是由一藍光或一過氧化氫所導致。
- 如申請專利範圍第2項所述的用途,其中該黑色素生成是由一藍光所導致。
- 如申請專利範圍第1項所述的用途,其中該玫瑰花萃取物的有效濃度為至少0.25%(v/v)。
- 如申請專利範圍第1項所述的用途,其中該萃取是於一介於65~85℃的溫度進行。
- 如申請專利範圍第1項所述的用途,其中該溶劑與該玫瑰花的體積比介於5~20:1~5。
- 如申請專利範圍第1項所述之用途,其中該組成物是一醫藥品、一食品產品或一保養品。
- 如申請專利範圍第9項所述的用途,其中該醫藥品包含一醫藥上可接受的載劑。
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TW201129325A (en) * | 2010-02-23 | 2011-09-01 | Yong-Zhen Chen | Method for making herbal enzyme |
CN101831354A (zh) * | 2010-04-22 | 2010-09-15 | 云南玫瑰庄园酒业有限责任公司 | 鲜玫瑰萃取液的提取方法 |
TW201509438A (zh) * | 2013-09-06 | 2015-03-16 | Li-Ren Wang | 促進毛髮增生之植物組成物 |
CN104367527A (zh) * | 2014-10-23 | 2015-02-25 | 青岛克立克信息技术有限公司 | 一种高效去角质面膜 |
CN105154278A (zh) * | 2015-07-23 | 2015-12-16 | 金华职业技术学院 | 一种佛手柑橘酒及其发酵方法 |
CN105250180B (zh) * | 2015-11-09 | 2018-11-27 | 北京工商大学 | 一种玫瑰鲜活水及其制备方法与应用 |
CN106306934A (zh) * | 2016-08-13 | 2017-01-11 | 安庆市老杨埠农业科技有限公司 | 一种发酵型佛手柑金珠果复合饮料及其制备工艺 |
CN106474016A (zh) * | 2016-12-16 | 2017-03-08 | 深圳市聚华太科技有限公司 | 一种头皮外用组合物及头皮滋养液 |
CN108175723A (zh) * | 2017-12-11 | 2018-06-19 | 百朗德生物化学(海门)有限公司 | 具有美白和防止皮肤老化的组合物及其在化妆品中的应用 |
CN108485846A (zh) * | 2018-03-22 | 2018-09-04 | 辽宁法奈迪生物工程有限公司 | 一种天然保湿抗氧化的洗脸皂及制备方法 |
CN109394850B (zh) * | 2018-12-17 | 2021-06-29 | 温州古木生物科技有限公司 | 一种具有延缓皮肤老化功效的中药组合物及其水提物和发酵物及应用 |
CN110592151B (zh) * | 2019-09-16 | 2020-06-30 | 哈尔滨美华生物技术股份有限公司 | 乳酸菌生产高含γ-氨基丁酸发酵液及其在化妆品中的应用 |
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