TWI621447B - 冰花癒傷組織萃取物於延緩皮膚細胞老化、調理皮膚、預防及治療皮膚癌之用途 - Google Patents
冰花癒傷組織萃取物於延緩皮膚細胞老化、調理皮膚、預防及治療皮膚癌之用途 Download PDFInfo
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Abstract
一種使用冰花(Mesembryanthemum crystallinum L.)癒傷組織萃取物於製造藥劑或護膚產品的用途,其中該藥劑或護膚產品係用於以下至少一者:延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌、及預防皮膚癌。
Description
本發明係關於冰花(Mesembryanthemum crystallinum L.)癒傷組織萃取物及其應用,尤其係關於使用冰花癒傷組織萃取物以延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌、及/或預防皮膚癌的應用。
紫外線(UV)是造成皮膚老化及病變的主要因素之一,其依波長可分為長波紫外線(UVA)、中波紫外線(UVB)、及短波紫外線(UVC)。已知,日光中的紫外線有超過90%是UVA,其具很強的穿透力,可穿透皮膚的真皮層而對皮膚造成傷害,經常暴露於UVA輻射下將引起皮膚光老化(photoaging)、活性氧分子(ROS)生成、DNA破壞、膠原蛋白流失、皮膚鬆弛等,甚至可能造成皮膚癌。
光老化是指皮膚長期暴露於紫外線輻射下,使得皮膚細胞加速老化,導致皮膚發生角質增厚、皮膚乾燥脫屑、產生細紋及斑點、皮膚鬆弛等現象。ROS的生成則會造成細胞中氧化
壓力升高、加速細胞老化,甚至造成細胞中細胞基質降解酵素的過度活化,進而導致膠原蛋白流失及皮膚鬆弛。此外,UVA亦可能破壞細胞中的DNA、造成DNA受損,過多受損DNA的累積不僅造成細胞老化,也可能使細胞癌化進而導致皮膚癌。
在皮膚癌中,以黑色素細胞瘤的致死率為最高。目前,臨床上主要是以手術切除的方式治療黑色素細胞瘤,至於無法承受手術治療的患者,則尚無有效的治療或預防方法。
有鑒於上述問題,業界仍亟需可有效延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌及/或預防皮膚癌的方法。本案發明人研究發現,冰花癒傷組織萃取物可有效降低紫外線對細胞所造成的傷害,具有降低皮膚紋理、毛孔、及經皮水分散失度的效果,且可有效誘發黑色素細胞瘤細胞之細胞凋亡,從而可用於延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌及/或預防皮膚癌。
本發明之一目的,在於提供一種使用冰花癒傷組織萃取物於製造護膚產品之用途,其中該護膚產品係用於以下至少一者:延緩皮膚細胞老化、調理皮膚、及修補皮膚。較佳地,該護膚產品係用於以下至少一者:抗光老化、抗氧化、及修復DNA,或用於以下至少一者:減緩膠原蛋白分解、減緩膠原蛋白流失、及減少肌膚紋理。
本發明之另一目的,在於提供一種使用冰花癒傷組織萃取物於製造藥劑之用途,其中該藥劑係用於以下至少一者:治療皮膚癌、預防皮膚癌,該皮膚癌可以為例如黑色素細胞瘤。
本發明之又一目的,在於提供一種用於延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌、及預防皮膚癌之至少一者的方法,其係包含對一有需要之個體投予一有效量之冰花癒傷組織萃取物。
第1圖係以DCF-DA染劑測試人類皮膚纖維母細胞中ROS含量的結果;第2圖係以qRT-PCR(real-time quantitative polymerase chain reaction)檢測人類皮膚纖維母細胞中CAT-1及SOD2-1基因表現量的結果;第3A至3C圖係以qRT-PCR檢測人類皮膚纖維母細胞中BER相關基因(包括UNG-1、OGG1-1、MPG-1、及APE1-1)、NER相關基因(包括ERCC1-1、ERCC6-1、及XPA-1)、及DSB相關基因(包括XRCC1-1及XRCC5-1)之表現量的結果;第4A及4B圖係以單細胞電泳檢測細胞之DNA受損情形的結果;第5圖係以qRT-PCR檢測人類皮膚纖維母細胞中TIMP1及COL1A1基因表現量的結果;第6圖係以MTT檢定法檢測人類皮膚纖維母細胞之存活率的結果;第7圖係以Skin analyzer TD膚質檢測儀檢測皮膚紋理的結果;以及第8圖係以MTT檢定法檢測黑色素瘤細胞之存活率的結果。
以下將描述根據本發明之部分具體實施態樣;惟,在不背離本發明精神下,本發明尚可以多種不同形式之態樣來實踐,不應將本發明保護範圍解釋為限於說明書所陳述者。此外,除非文中有另外說明,於本說明書中(尤其是在後述專利申請範圍中)所使用之「一」、「該」及類似用語應理解為包含單數及複數形式;所謂“有效量”,是指投予至個體時,可有效至少部分改善懷疑個體的病情的化合物數量;所謂“個體”是指人類或非人的哺乳動物;所謂“治療”是包括預防特定疾病或症狀、減輕特定的疾病或症狀及/或防止或消除該病症。
冰花(Mesembryanthemum crystallinum L.)在分類學上屬於番杏科、松葉菊屬的雙子葉一年生植物。本案發明人研究發現,冰花植株在受傷後所產生之癒傷組織具有類似哺乳類全能幹細胞的特性,該癒傷組織萃取物可有效抗光老化、抗氧化、修復DNA、減緩膠原蛋白分解、減緩膠原蛋白流失、及減少肌膚紋理。
因此,本發明係提供一種使用冰花癒傷組織萃取物於延緩皮膚細胞老化、調理皮膚、修補皮膚、預防皮膚癌、及/或治療皮膚癌之應用。前述應用包括:使用冰花癒傷組織萃取物於製造一延緩皮膚細胞老化、調理皮膚、及/或修補皮膚之護膚產品;使用冰花癒傷組織萃取物於製造一治療皮膚癌及/或預防皮膚癌之藥劑;以及使用冰花癒傷組織萃取物以對有需要之個體提供延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌、及預防皮膚癌之至少一者的方法。
本發明所採用之冰花癒傷組織萃取物,係可經由以溶劑萃取冰花癒傷組織而提供。其中,所採用之溶劑可以為極性溶劑,例如醇、水、或其組合。較佳地,該溶劑係選自C1-4醇、水、及其組合。舉例言之,可以乙醇進行該萃取操作。視需要地,可於超音波震盪操作下進行萃取,以提升萃取效果。較佳地,可以在進行萃取之前,先進行一乾燥處理。
於本發明之部分實施態樣中,係於進行萃取之前,先冷凍乾燥冰花癒傷組織。舉例言之,可以100:1(水:經冷凍乾燥之冰花癒傷組織)之重量比混合水與經冷凍乾燥之冰花癒傷組織,並於70℃下對所得到之混合物進行超音波震盪,歷時45分鐘以完成萃取。或者,可以10:1之重量比混合70%乙醇與經冷凍乾燥冰花癒傷組織,並在70℃下對所得到之該混合物進行超音波震盪,歷時30分鐘以完成萃取。
該冰花癒傷組織可透過包含如下步驟之操作提供:I. 以6%次氯酸鈉溶液沖洗冰花植株,其後以滅菌水沖洗冰花植株,視需要地,可重複前述沖洗操作;II. 在冰花植株的表面上切出傷口,以誘導產生癒傷組織(歷時1至3個月);III. 在25℃、50至60%濕度下,以MS培養基(Murashige and Skoog Basal Medium,購自Sigma公司,產品編號:M5519)培養步驟II所得之癒傷組織(歷時1至1.5個月)。
於本發明中,冰花癒傷組織萃取物可以萃取所得之萃取原液直接使用,或視需要對該萃取原液進行例如過濾、滅菌、濃縮、稀釋等一或多個步驟,以提升萃取液之使用便利性。舉例
言之,透過例如濃縮乾固、噴霧乾燥、或冷凍乾燥等操作,可由萃取液得到方便攜帶或方便儲存之粉末產物。
根據本發明所提供之護膚產品係可呈任何合宜的形式,並無特殊的限制,端視所欲之用途而呈對應之合宜劑型。舉例言之,但不以此為限,該護膚產品可呈供直接外用之乳液、乳霜、凝膠(例如水凝膠)、膏狀物(例如分散膏、軟膏)、噴霧劑、或溶液(例如洗液、懸浮液)等形式。或者,可將本發明之護膚產品製備成可供吞食或飲用的食品形式,例如健康食品、美容飲品等。此外,亦可將本發明護膚產品以皮下注射之針劑形式提供。
類似地,根據本發明所提供之藥劑係可呈任何合宜的型式,並無特殊限制,端視所欲之用途而呈對應之合宜劑型。舉例言之,但不以此為限,該藥劑可以口服或非經口服(例如:經皮、皮下、靜脈內、肌肉、腹腔、或鼻腔)之投藥方式施用至有需要之個體上。其中,視使用形式及用途而定,可選用合宜之載劑以提供該醫藥組合物或藥物,其中,該載劑包括賦形劑、稀釋劑、輔助劑、安定劑、吸收延遲劑、崩散劑、增溶劑、乳化劑、抗氧化劑、黏合劑、結合劑、增黏劑、分散劑、懸浮化劑、潤滑劑、吸濕劑等。
以適於口服投藥之藥劑形式為例,該藥劑中可含有任何不會不利影響冰花癒傷組織萃取物及/或其活性成分之所欲效益的醫藥上可接受之載劑,例如:水、食鹽水、葡萄糖(dextrose)、甘油、乙醇或其類似物、纖維素、澱粉、糖膨潤土(sugar bentonite)、及前述之組合。可利用任何合宜之方法,以適於口服投藥的劑型
提供該藥劑,例如:錠劑(例如糖衣錠)、丸劑、膠囊劑、顆粒劑、散劑、流浸膏劑、溶液劑、糖漿劑、懸液劑、酊劑等。
至於適於皮下靜脈內、肌肉、或腹腔注射之針劑或點滴劑型,則可於該藥劑中含有一或多種例如等張溶液、鹽類緩衝液(如磷酸鹽緩衝液或檸檬酸鹽緩衝液)、增溶劑、乳化劑、5%糖溶液、以及其他載劑等成分,以靜脈輸注液、乳劑靜脈輸注液、乾粉注射劑、懸液注射劑、或乾粉懸液注射劑等劑型提供該藥劑。或者,將該藥劑製備成一注射前固體,以可溶於其他溶液或懸浮液中之劑型、或可乳化之劑型提供該注射前固體,並於投予至有需要之個體之前,將該注射前固體溶於其他溶液或懸浮液中或將其乳化,提供所欲之注射劑。
視需要地,可於根據本發明所提供之護膚產品或藥劑中另外含有合宜用量之添加劑,例如可提高該護膚產品或藥劑於服用時的口適感及視覺感受之調味劑、調色劑、著色劑等,以及可改善該護膚產品或藥劑的穩定性及儲存性之緩衝劑、保存劑、防腐劑、抗菌劑、抗真菌劑等。此外,該護膚產品或藥劑可視需要另含一或多種其他活性成分(例如,玻尿酸、杏仁酸、熊果素、膠原蛋白、彈性蛋白等),或者與含該一或多種其他活性成分之護膚產品或藥物併用,以進一步加強該護膚產品或藥劑之功效或增加製劑配方的運用靈活性與調配度。原則上,只要所添加的其它成分或添加劑不會對本發明護膚產品或藥劑之所欲功效產生不利的影響即可。
於根據本發明之應用所提供之護膚產品或藥劑係可
以一日一次、一日多次、或數日一次等不同頻率施用,端視投予個體之需求、年齡、體重、健康況狀、及施用目的而異。
當施用根據本發明所提供之護膚產品於皮膚表面以延緩皮膚細胞老化、調理皮膚、及/或修補皮膚時,可視產品型態而含有不同濃度之冰花癒傷組織萃取物。舉例言之,當以精華液之形式提供本發明冰花癒傷組織萃取物時,該精華液中之冰花癒傷組織萃取物之濃度可以為例如0.01-10重量%(如:1重量%)。而當以美容飲品形式提供本發明冰花癒傷組織萃取物時,該飲品中之冰花癒傷組織萃取物之濃度則可以為例如1-1000ppm(如:100ppm)。
本發明亦提供一種用於延緩皮膚細胞老化、調理皮膚、修補皮膚、治療皮膚癌、及預防皮膚癌之至少一者的方法,其係包含對一有需要之個體投予一有效量之冰花癒傷組織萃取物。其中,有關該冰花癒傷組織萃取物的投予途徑、投予形式、適用劑量、以及相關治療或預防之應用,均如上述之說明。
茲以下列實施例進一步例示說明本發明。其中該等實施例僅提供作為說明,而非用以限制本發明之保護範圍。本發明保護範圍係如後附申請專利範圍所示。
實施例
實施例1:萃取物的製備
將購自台灣蔬菜之家園藝資材行之冰花種子(商品代碼:00K20)培育成冰花植株,並將該植株分成A、B兩批。其中,對A批施以如下處理,以提供冰花癒傷組織:I. 以6%次氯酸鈉溶液沖洗冰花植株,其後以滅菌水沖洗冰花植
株,視需要地,可重複前述沖洗操作;II. 在冰花植株的表面上切出傷口,以誘導產生癒傷組織(歷時1至3個月);III. 在25℃、50至60%濕度下,以MS培養基(Murashige and Skoog Basal Medium,購自Sigma公司,產品編號:M5519)培養步驟II所得之癒傷組織(歷時1至1.5個月)。
分別以下列步驟處理上述第B批之冰花植株以及由第A批冰花植株所得之冰花癒傷組織,以製備冰花植株(IP)萃取物及冰花癒傷組織(IPC)萃取物:
(1)冰花植株(或冰花癒傷組織)置於-22℃下進行冷凍乾燥,歷時12小時;以及
(2)粉碎步驟(1)之經乾燥的冰花植株(或冰花癒傷組織),以提供冰花植株粉末(或冰花癒傷組織粉末)。
(3)將70%乙醇與步驟(2)之冰花植株粉末(或冰花癒傷組織粉末)以10:1(乙醇:粉末)的重量比混合;
(4)在70℃下,對步驟(3)所獲得的混合物進行超音波震盪,歷時30分鐘;
(5)以濾膜過濾步驟(4)所得之混合物,以提供一濾液;
(6)加熱由步驟(5)所獲得之濾液至95℃,並維持20分鐘,以進行滅菌;
(7)待步驟(6)之濾液冷卻後,將其冷藏保存以供後續實驗使用。
實施例2:冰花癒傷組織(IPC)萃取物之抗氧化功效
於MEM培養基(購自Gibco,產品編號:61100-061)中培養人類皮膚纖維母細胞(CCD-966SK;購自ATCC)歷時24小時,其後,將細胞分為四組,並進行以下處理:
(1)控制組:將細胞置於MEM培養基中培養2小時(即,將細胞培養於不含IP萃取物及IPC萃取物的培養液中)。
(2)H2O2組:將細胞置於MEM培養基中培養1小時(即,將細胞培養於不含IP萃取物及IPC萃取物的培養液中),接著,加入H2O2(使其於培養基中的最終濃度達到1毫莫耳濃度)進行處理,歷時1小時。
(3)H2O2+IP組:將細胞置於含有2毫克/毫升得自實施例1的冰花植株(IP)萃取物的MEM培養基中培養1小時,接著,加入H2O2(使其於培養基中的最終濃度達到1毫莫耳濃度)進行處理,歷時1小時。
(4)H2O2+IPC組:將細胞置於含有2毫克/毫升得自實施例1的冰花癒傷組織(IPC)萃取物的MEM培養基中培養1小時,接著,加入H2O2(使其於培養基中的最終濃度達到1毫莫耳濃度)進行處理,歷時1小時。
接著,以DCF-DA染劑(購自Sigma公司,產品型號:D6883)處理上述各組細胞歷時15分鐘,其後以流式細胞儀偵測各組之螢光強度,其中,由於ROS可將DCF-DA(不具螢光)轉變為
DCF(具有螢光),故所測得之螢光強度可代表細胞中的ROS含量,螢光強度越高代表細胞中的ROS含量越高。最後,以控制組的結果為基準,計算其他各組的相對螢光強度。結果示於第1圖(*表示與H2O2組有顯著差異,p<0.05;**表示與H2O2組有顯著差異,p<0.01;#表示與H2O2+IP組有顯著差異,p<0.05)。
由第1圖可知,相較於控制組,H2O2組的螢光強度顯著上升。然而,相較於H2O2組,H2O2+IP組與H2O2+IPC組的螢光強度皆明顯下降。此外,相較於H2O2+IP組,H2O2+IPC組的螢光強度明顯更低。前述結果顯示,冰花癒傷組織(IPC)萃取物可有效抵抗H2O2氧化壓力、降低細胞中的ROS含量,可用於抗氧化,而且所提供之效果明顯優於冰花植株(IP)萃取物。
實施例3:冰花癒傷組織萃取物於延緩皮膚細胞老化、調理皮膚、及修補皮膚的效果
(3-1)人類皮膚纖維母細胞之處理
於MEM培養基中培養人類皮膚纖維母細胞(CCD-966SK;購自ATCC)歷時24小時,其後,將細胞分為四組,並進行以下處理:
(1)控制組:將細胞置於MEM培養基中培養48小時(即,將細胞培養於不含冰花癒傷組織萃取物的培養液中)。
(2)UVA+6hr組、UVA+24hr組、UVA+48hr組:分別將細胞置於含有2毫克/毫升得自實施例1的冰花癒傷組織萃取物的MEM培養基中培養6小時、24小時、及48小時。
(3)接著,以UVA照射上述各組細胞,歷時1小時。
(3-2)抗光老化及抗氧化
為進一步了解本發明冰花癒傷組織萃取物於抗光老化及抗氧化的效果,於完成實施例3-1之後,進行qRT-PCR,以檢測各組細胞中CAT-1及SOD2-1等抗氧化基因的表現量,並以控制組的結果為基準,計算其他組的相對基因表現量。結果示於第2圖。
由第2圖可知,相較於控制組(未經冰花癒傷組織萃取物處理),經冰花癒傷組織萃取物處理之組別(包括UVA+6hr組UVA+24hr組、及UVA+48hr組)的抗氧化基因(即,CAT-1及SOD2-1)表現量皆明顯提升。此結果顯示,冰花癒傷組織萃取物可提供抗光老化及抗氧化的效果,故可用於延緩皮膚細胞老化。
(3-3)修復DNA
生物體中具有鹼基切除修復(base excision repair,BER)、核苷酸切除修復(nucleotide excision repair,NER)、雙股斷裂末端接合(double-strand break end joint,DSB)等DNA修復機制,故可避免因為DNA突變或斷裂所造成的傷害。
為了解本發明冰花癒傷組織萃取物是否可修復DNA,於完成實施例3-1之後所進行之qRT-PCR中,除檢測各組細胞中CAT-1及SOD2-1等抗氧化基因的表現量以外,並檢測細胞中有關BER、NER、及DSB等機制的基因表現量,後者之結果示於第3A至3C圖(*p<0.05、**p<0.01、***p<0.001)。
由第3A至3C圖可知,相較於控制組,經冰花癒傷組織萃取物處理之組別(包括UVA+6hr組UVA+24hr組、及UVA+48hr組)的BER相關基因(包括UNG-1、OGG1-1、MPG-1、及APE1-1)、NER相關基因(包括ERCC1-1、ERCC6-1、及XPA-1)、及DSB相關
基因(包括XRCC1-1及XRCC5-1)表現量皆明顯提升。前述結果顯示,冰花癒傷組織萃取物可有效修復DNA,並藉此達到延緩皮膚細胞老化的效果。
(3-4)保護DNA
已知,在單細胞電泳檢測中,DNA受損的細胞會出現拖尾的現象,因此,可藉由計算有拖尾現象之細胞的比例來評估DNA受損程度。
為進一步了解本發明冰花癒傷組織萃取物是否可保護細胞之DNA,係對實施例2之控制組、H2O2組、及H2O2+IPC組三組細胞進行單細胞電泳檢測。結果示於第4A及4B圖(** p<0.01)。
由第4A及4B圖可知,相較於控制組,H2O2組中有拖尾現象(即,DNA受損)的細胞比例明顯較高。然而,相較於H2O2組,H2O2+IPC組中有拖尾現象的細胞比例則明顯較低。前述結果顯示,冰花癒傷組織萃取物可有效保護細胞DNA,並藉此達到延緩皮膚細胞老化的效果。
(3-5)減緩膠原蛋白分解及流失
膠原蛋白分解及流失會加速皮膚老化,其中,已知TIMP1的基因表現量上升代表膠原蛋白的分解被抑制,COL1A1表現上升,代表膠原蛋白表現量增加。為了解本發明冰花癒傷組織萃取物是否可減緩皮膚之膠原蛋白分解及流失,係於完成實施例3-1之後所進行之qRT-PCR中,亦檢測控制組及UVA+24hr組細胞中TIMP1及COL1A1的表現量,並以控制組作為基準計算UVA+24hr組的相對基因表現量。結果示於第5圖(*p<0.05、**p<0.01)。
由第5圖可知,相較於控制組,UVA+24hr組的TIMP1
及COL1A1表現量皆明顯提升。此結果顯示,冰花癒傷組織萃取物可有效降低膠原蛋白分解及提升膠原蛋白合成,故可用於減緩膠原蛋白分解及/或流失,達到調理皮膚、及修補皮膚的效果。
(3-6)提高人類皮膚纖維母細胞存活率
於人類皮膚纖維母細胞穩定培養24小時後,將其分為六組,並進行以下處理:
(1)控制組:將細胞置於MEM培養基中培養24小時(即,將細胞培養於不含冰花癒傷組織萃取物的培養液中)。
(2)UVA組:將細胞置於MEM培養基中培養24小時(即,將細胞培養於不含冰花癒傷組織萃取物的培養液中),接著,以UVA照射細胞,歷時1小時。
(3)UVA+0.75組、UVA+1組、UVA+1.5組、及UVA+2組:分別將細胞置於含有0.75、1、1.5、及2毫克/毫升由實施例1獲得之冰花癒傷組織萃取物的MEM培養基中培養24小時,接著,以UVA照射細胞,歷時1小時。
其後,以MTT檢定法檢測各組細胞之存活率。結果示於第6A及6B圖(** p<0.01)。由第6A圖可知,相較於控制組,UVA組之細胞存活率明顯較低。然而,由第6B圖可知,相較於UVA組,經冰花癒傷組織萃取物處理之組別(包括UVA+0.75組、UVA+1組、UVA+1.5組、及UVA+2組)的細胞存活率皆顯著回升,且冰花癒傷組織萃取物之使用濃度係與細胞存活率成正比。前述結果顯示,冰花癒傷組織萃取物可有效降低UVA對細胞的傷害,且在試驗濃度範圍內對正常人類皮膚纖維母細胞不具毒性。
(3-7)減少肌膚紋理
本實驗採自身對照方式進行,10位自願受試者在第0週時,以Skin analyzer TD膚質檢測儀及DermaLab® Combo超音波膚質分析儀測試並記錄自身之皮膚紋理、毛孔、及經皮水分散失度;接著,每位受試者早晚各一次以精華液(含有1重量%之由實施例1獲得之冰花癒傷組織萃取物)塗抹半臉,並以安慰劑(即,成分皆與精華液相同,但不含本發明之冰花癒傷組織萃取物)塗抹另半臉,持續6週;其後,測試並記錄使用精華液(含有本發明之冰花癒傷組織萃取物)或安慰劑6週後的皮膚紋理(即,細紋)、毛孔、及經皮水分散失度。結果示於表1及第7圖。
如表1所示,在使用含有本發明冰花癒傷組織萃取物之精華液6週後,受試者的皮膚紋理明顯減少、毛孔明顯縮小,且經皮水分散失度明顯降低。此外,如第7圖顯示,相較於塗抹安慰劑,受試者在塗抹精華液(含有本發明之冰花癒傷組織萃取物)6週後,眼部周圍的細紋明顯改善。前述結果顯示,本發明之冰花癒傷組織萃取物確實具有調理皮膚及修補皮膚的效果。
實施例4:冰花癒傷組織萃取物於治療及預防黑色素
細胞瘤的效果
於黑色素瘤細胞(購自ATCC,產品編號:CRL-6475)穩定培養24小時後,將其分為六組,其中一組係繼續培養於不含冰花癒傷組織萃取物的DMEM培養基(購自Gibco,產品編號:12100-046)中,歷時48小時(此為「控制組」),其餘五組則分別培養於含有由實施例1獲得之冰花癒傷組織萃取物的DMEM培養基中(冰花癒傷組織萃取物於培養基中的最終濃度分別為0.25、0.5、1、2及4毫克/毫升),歷時48小時。接著,以MTT檢定法檢測各組的存活率,並以控制組之結果為基準,計算其他各組之相對細胞存活率。結果示於第8圖(** p<0.01)。
如第8圖所示,當冰花癒傷組織萃取物的濃度高於0.5毫克/毫升時,黑色素瘤細胞的存活率係與冰花癒傷組織萃取物的使用濃度呈反比。此結果顯示,冰花癒傷組織萃取物可有效毒殺黑色素瘤細胞,故可用於治療及/或預防皮膚癌。
Claims (3)
- 一種使用冰花癒傷組織萃取物於製造一藥劑之用途,其中該藥劑係用於治療皮膚癌。
- 如請求項1之用途,其中該藥劑係用於治療黑色素細胞瘤。
- 如請求項1或2中任一項之用途,其中該藥劑係以經皮或口服之投藥方式施用。
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