TWI827334B - Use of rutin in preparing compositions for promoting hair growth - Google Patents
Use of rutin in preparing compositions for promoting hair growth Download PDFInfo
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- TWI827334B TWI827334B TW111141869A TW111141869A TWI827334B TW I827334 B TWI827334 B TW I827334B TW 111141869 A TW111141869 A TW 111141869A TW 111141869 A TW111141869 A TW 111141869A TW I827334 B TWI827334 B TW I827334B
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Abstract
一種蘆丁用於製備促進生髮組成物的用途，該促進生髮組成物含有該蘆丁成分，並以一有效量外用施予在一個體的脫毛部位皮膚上，能夠促進該個體的脫毛部位之毛囊細胞生長，其中該促進生髮組成物當中該蘆丁成分為天然植物的萃取物，在皮膚使用上無副作用產生，並可使得該個體的脫毛部位能明顯看出皮膚毛髮增生，藉以有效提供優異促進毛髮生長及預防脫髮的效果。A use of rutin for preparing a hair growth-promoting composition. The hair growth-promoting composition contains the rutin component and is externally applied to the skin of a person's hair removal site in an effective amount, which can promote the hair follicles of the individual's hair removal site. Cell growth, wherein the rutin component of the hair growth-promoting composition is a natural plant extract, has no side effects on the skin, and can make the individual's hair removal area clearly visible skin hair growth, thereby effectively providing excellent promotion Hair growth and hair loss prevention effects.
Description
本發明係一種天然蘆丁萃取物的用途；特別是指一種蘆丁用於製備促進生髮組成物的用途。The present invention relates to the use of natural rutin extract; in particular, it refers to the use of rutin in preparing a composition for promoting hair growth.
在現今高壓社會環境中，繁忙的課業及事業逐漸影響各年齡層的身心健康，相對引發各式慢性疾病，再加上近代生活習慣、環境、基因、藥物以及賀爾蒙問題，脫髮/掉髮的皮膚疾病已經廣泛分布於各年齡層。In today's high-pressure social environment, busy schoolwork and careers gradually affect the physical and mental health of all age groups, relatively causing various chronic diseases. Coupled with modern living habits, environment, genes, drugs and hormonal problems, hair loss/hair loss Skin diseases have been widely distributed among all age groups.
目前市面上推出多種預防及改善脫髮的生髮產品，莫過於外用落健minoxidil、口服柔沛finasteride，雖然前述生髮產品在臨床試驗上均具有優異地毛髮生長效果，但仍有接近半數的落健使用者無法取得良好的生髮效果，而柔沛的生髮效果則可能因為服用天數拉長反而降低效用。There are currently a variety of hair growth products on the market to prevent and improve hair loss, including topical Rogaine minoxidil and oral finasteride. Although the aforementioned hair growth products have excellent hair growth effects in clinical trials, there are still nearly half of Rogaine users who cannot A good hair growth effect is obtained, but the hair growth effect of Ropei may be reduced due to the prolonged use of it.
除此之外，前述生髮產品在使用時均會產生相對的副作用，具體來說，落健的外用塗敷有可能導致皮膚紅腫熱痛、血壓過低等副作用；而服用柔沛將導致過敏反應及雄性使用者性功能障礙；如此一來，對於許多使用者而言，會因副作用的現象而降低該生髮產品的使用意願。In addition, the aforementioned hair growth products will produce relative side effects when used. Specifically, the external application of Rogaine may cause side effects such as redness, swelling, heat and pain on the skin, and low blood pressure; while taking Rogaine will cause allergic reactions and Sexual dysfunction in male users; thus, for many users, the side effects will reduce their willingness to use the hair growth product.
有鑑於此，本發明之目的在於提供一種蘆丁用於製備促進生髮組成物的用途，該蘆丁成分為天然植物的萃取物，在皮膚使用上無副作用產生，還能有效活化毛囊細胞生長。In view of this, the object of the present invention is to provide a use of rutin for preparing a hair growth-promoting composition. The rutin component is a natural plant extract, has no side effects when used on the skin, and can effectively activate the growth of hair follicle cells.
緣以達成上述目的，本發明提供的蘆丁用於製備促進生髮組成物的用途，該促進生髮組成物含有該蘆丁成分，並以一有效量外用施予在一個體的脫毛部位皮膚上，能夠促進該個體的脫毛部位之毛囊細胞生長。In order to achieve the above object, the rutin provided by the present invention is used for preparing a hair growth promoting composition. The hair growth promoting composition contains the rutin component and is externally applied to the skin of a person's hair removal area in an effective amount. It can promote the growth of hair follicle cells in the individual's hair removal area.
本發明之效果在於，該促進生髮組成物當中該蘆丁成分為天然植物的萃取物，在皮膚使用上無副作用產生，且經過一系列動物試驗顯示，該個體在持續外用施予7~14天時，可使得該個體的脫毛部位能明顯看出皮膚毛髮增生，提供優異促進毛髮生長及預防脫髮的效果。The effect of the present invention is that the rutin component in the hair growth-promoting composition is a natural plant extract, which has no side effects when used on the skin, and a series of animal tests have shown that the individual can continue to apply it externally for 7 to 14 days. At this time, the skin and hair growth can be clearly seen in the individual's hair removal area, providing excellent effects in promoting hair growth and preventing hair loss.
本發明提供的蘆丁用於製備促進生髮組成物的用途，該促進生髮組成物含有該蘆丁成分（Rutin），並以一有效量外用施予在一個體的脫毛部位皮膚上，且在本實施例中，該蘆丁成分能夠活化該個體的脫毛部位當中皮膚組織的β-連環蛋白（β-catenin）之表現，藉以促進該個體的脫毛部位之毛囊細胞生長；更進一步說明，該蘆丁成分（Rutin）係為蕎麥植物之天然萃取物，該促進生髮組成物中含有一溶劑具有聚乙二醇-400及乙醇，該聚乙二醇-400與乙醇以7：3的體積比例進行調配，該蘆丁成分與該溶劑混合形成該促進生髮組成物，且該促進生髮組成物對該個體的脫毛部位持續外用施予7~14天時，該個體的脫毛部位受到該蘆丁成分的活化能明顯看出皮膚毛髮增生。The rutin provided by the present invention is used for preparing a hair growth promoting composition. The hair growth promoting composition contains the rutin component (Rutin) and is externally applied to the skin of a person's hair removal area in an effective amount, and in the present invention In embodiments, the rutin component can activate the expression of β-catenin in the skin tissue of the individual's hair removal area, thereby promoting the growth of hair follicle cells in the individual's hair removal area; further illustrating, the rutin The ingredient (Rutin) is a natural extract of the buckwheat plant. The hair growth-promoting composition contains a solvent containing polyethylene glycol-400 and ethanol. The polyethylene glycol-400 and ethanol are prepared in a volume ratio of 7:3. , the rutin component is mixed with the solvent to form the hair growth promoting composition, and when the hair growth promoting composition is continuously applied externally to the hair removal site of the individual for 7 to 14 days, the hair removal site of the individual is activated by the rutin component Skin and hair growth can be clearly seen.
本文中所載「有效量」（effective amount）在此處係指一組成物當中有效成分的用量足以產生欲求的生理反應，該有效成分的有效量不必然能夠治癒疾病或病徵，但能夠延緩、阻礙或防止病徵的發生，或是可緩減病徵的發生；該有效成分的具體有效量取決於多種因素，例如欲試驗的特定狀況、個體的生理條件(如，個體重、年齡或性別)、接受試驗的個體類型及持續時間的本質，以及所用的具體配方。舉例來說，可將該有效成分的有效量表示成重量，譬如以克、毫克或微克來表示；或者將該有效成分的有效量表示成重量相對於體重的比例，例如為每公斤體重多少毫克（mg/kg）。或者是，可將該有效成分的有效量表示成濃度，例如莫耳濃度、重量濃度、體積濃度、重量莫耳濃度、莫耳分率、重量分率及混合比值，且習知技藝人士可依據動物模式的試驗劑量來換算成人體等效劑量（human equivalent dose,HED）。The "effective amount" as used herein refers to the amount of active ingredient in a composition that is sufficient to produce the desired physiological response. The effective amount of the active ingredient does not necessarily cure the disease or symptoms, but can delay, Hinder or prevent the occurrence of symptoms, or can alleviate the occurrence of symptoms; the specific effective amount of the active ingredient depends on many factors, such as the specific condition to be tested, the individual's physiological conditions (such as individual weight, age or gender), The nature of the types and duration of individuals tested, and the specific formulation used. For example, the effective amount of the active ingredient can be expressed as weight, such as grams, milligrams or micrograms; or the effective amount of the active ingredient can be expressed as a ratio of weight to body weight, such as milligrams per kilogram of body weight. (mg/kg). Alternatively, the effective amount of the active ingredient can be expressed as a concentration, such as molar concentration, weight concentration, volume concentration, weight molar concentration, molar fraction, weight fraction and mixing ratio, and those skilled in the art can The experimental dose in animal models is converted into human equivalent dose (HED).
在本實施例中，該蘆丁成分使用所述有效量是以莫耳濃度或重量百分比作為表示，其中當該蘆丁成分的有效量以莫耳濃度表示時，該蘆丁成分使用所述有效量係介於10至60mM，在一較佳實施例中，該蘆丁成分使用所述有效量係介於12.5至50mM，在另一較佳實施例中，該蘆丁成分使用所述有效量係介於25至50mM；另外，當該蘆丁成分的有效量以重量百分比表示時，該蘆丁成分使用所述有效量係介於0.5~4wt%，在一較佳實施例中，該蘆丁成分使用所述有效量係介於0.76~3.05wt%，在另一較佳實施例中，該蘆丁成分使用所述有效量係介於1.53~3.05wt%。In this embodiment, the effective amount of the rutin component is expressed in molar concentration or weight percentage. When the effective amount of the rutin component is expressed in molar concentration, the effective amount of the rutin component is expressed in molar concentration or weight percentage. The amount is between 10 and 60mM. In a preferred embodiment, the effective amount of the rutin component is between 12.5 and 50mM. In another preferred embodiment, the effective amount of the rutin component is used. is between 25 and 50mM; in addition, when the effective amount of the rutin component is expressed in weight percent, the effective amount of the rutin component is between 0.5 and 4wt%. In a preferred embodiment, the rutin component is The effective amount of the rutin component ranges from 0.76 to 3.05wt%. In another preferred embodiment, the effective amount of the rutin component ranges from 1.53 to 3.05wt%.
特別說明地，本實施例中該蘆丁成分使用所述有效量經篩選具有優異促進毛髮生長及預防脫髮的效果，其中該蘆丁成分使用所述有效量低於10mM，或者低於0.5wt%時，該蘆丁成分因有效量過少而使毛髮生長效果不佳；當該蘆丁成分使用所述有效量高於60mM，或者高於4wt%時，則因該蘆丁成分無法完全溶解於該溶劑，可能會導致後續在外用施予過程無法均勻塗佈在該個體的脫毛部位，致使影響毛髮生長效益。Specifically, in this embodiment, the effective amount of the rutin component used has been screened to have excellent effects on promoting hair growth and preventing hair loss, wherein the effective amount of the rutin component used is less than 10mM, or less than 0.5wt% When the effective amount of the rutin component is too small, the hair growth effect is poor; when the effective amount of the rutin component is higher than 60mM, or higher than 4wt%, the rutin component cannot be completely dissolved in the Solvents may cause the subsequent external application process to be unable to be evenly coated on the hair removal site of the individual, thus affecting the hair growth effect.
本文中所載「個體」（subject），係指包含人類等可接受本實施該促進生髮組成物的動物，除非另有所指，否則「個體」一詞意指包含任何在接受本實施該促進生髮組成物的動物後，可產生治療效益的哺乳動物，例如人類、大鼠、小鼠、天竺鼠、猴子、豬、山羊、牛、馬、狗、貓、鳥及家禽。在本實施例是以小鼠作為「個體」試驗。The term "subject" as used herein refers to animals including humans and other animals that can receive the hair growth-promoting composition of the present invention. Unless otherwise specified, the term "subject" means any animal that accepts the hair-promoting composition of the present invention. The therapeutic benefit may be produced in mammals such as humans, rats, mice, guinea pigs, monkeys, pigs, goats, cattle, horses, dogs, cats, birds and poultry. In this example, mice were used as "individual" tests.
為充分瞭解本發明之目的、特徵及功效，本實施例茲提供該促進生髮組成物的動物實驗模型、於形態學觀察毛髮變化、於毛髮長度分析，以及於組織學觀察毛囊變化，藉以說明本實施該促進生髮組成物當中蘆丁成分確實提供促進毛髮生長的效果。下述具體實施例可進一步證明本發明可實際應用範圍，且任何所屬技術領域中具有通常知識者在參照本發明之後，當可對其參數或設定作適當的更動，惟其仍應屬於本發明之範疇內。In order to fully understand the purpose, characteristics and efficacy of the present invention, this embodiment provides an animal experimental model of the hair growth-promoting composition, morphological observation of hair changes, hair length analysis, and histological observation of hair follicle changes to illustrate the present invention. The rutin component in the hair growth promoting composition can indeed provide the effect of promoting hair growth. The following specific examples can further prove the scope of practical application of the present invention, and any person with ordinary knowledge in the technical field can make appropriate changes to the parameters or settings after referring to the present invention, but they should still fall within the scope of the present invention. within the scope.
建立毛髮增生動物實驗模型：Establish an animal experimental model of hair hyperplasia:
本實施例所選用的「個體」係使用由國家動物實驗中心購買週齡7~8週之C57BL/6小鼠進行試驗，各該個體係先於環境中適應一週後再進行後續各項試驗，其中該個體的脫髮部位係在背部，有利於後續觀察。The "individuals" selected in this example were 7-8 week old C57BL/6 mice purchased from the National Animal Experiment Center for testing. Each system was adapted to the environment for a week before subsequent tests were conducted. The individual's hair loss area is tied to the back, which is convenient for follow-up observation.
本試驗之組別分別包含控制組及實驗組1~3：The groups of this experiment include control group and experimental group 1~3 respectively:
控制組：係使用前述溶劑具有該聚乙二醇-400及該乙醇，試驗過程係將該溶劑外用塗敷在該個體的脫髮部位持續14天。Control group: The aforementioned solvent containing the polyethylene glycol-400 and the ethanol was used. The test process was to apply the solvent externally to the hair loss area of the individual for 14 days.
實驗組1：係使用該促進生髮組成物外用塗敷在該個體的脫髮部位持續14天，其中該促進生髮組成物當中該蘆丁成分使用的有效量以莫耳濃度計係為12.5mM；若該蘆丁成分使用的有效量以重量百分比計係為0.76wt%。Experimental Group 1: The hair growth-promoting composition was applied externally to the hair loss area of the individual for 14 days, and the effective amount of the rutin component in the hair growth-promoting composition was 12.5mM in molar concentration; if The effective amount of the rutin component used is 0.76wt% in terms of weight percentage.
實驗組2：係使用該促進生髮組成物外用塗敷在該個體的脫髮部位持續14天，其中該促進生髮組成物當中該蘆丁成分使用的有效量以莫耳濃度計係為25mM；若該蘆丁成分使用的有效量以重量百分比計係為1.53wt%。Experimental Group 2: The hair growth-promoting composition was applied externally to the hair loss area of the individual for 14 days, and the effective amount of the rutin component in the hair growth-promoting composition was 25mM in molar concentration; if the The effective amount of rutin used is 1.53wt% in terms of weight percentage.
實驗組3：係使用該促進生髮組成物外用塗敷在該個體的脫髮部位持續14天，其中該促進生髮組成物當中該蘆丁成分使用的有效量以莫耳濃度計係為50mM；若該蘆丁成分使用的有效量以重量百分比計係為3.05wt%。Experimental Group 3: The hair growth-promoting composition was applied externally to the hair loss area of the individual for 14 days, and the effective amount of the rutin component in the hair growth-promoting composition was 50mM in terms of molar concentration; if the The effective amount of rutin used is 3.05wt% in terms of weight percentage.
觀察該個體（小鼠）毛髮生長：Observe the hair growth of the individual (mouse):
由於週齡7~8週之 C57BL/6小鼠其毛髮週期大部分處於休止期，因而在本實驗係將該控制組及該實驗組1~3進行生髮實驗，將刺激該個體處於休止期的毛囊產生毛髮週期且重新進入生長期，其中實驗過程該控制組及該實驗組1~3分別在第1、7、14天的實驗期間使用相機進行背部皮膚影像紀錄的操作。Since the hair cycle of C57BL/6 mice aged 7 to 8 weeks is mostly in the telogen phase, in this experiment, the control group and the experimental group 1 to 3 were used for hair growth experiments to stimulate the hair growth of the individuals in the telogen phase. The hair follicles generate a hair cycle and re-enter the growth phase. During the experimental process, the control group and the experimental groups 1 to 3 used cameras to record images of the back skin on the 1st, 7th, and 14th days of the experiment respectively.
請參閱圖1觀察結果來看，該控制組及該實驗組1~3在第1天時能明顯看到各該個體背部上的脫毛部位；在第7天時，控制組當中該個體的脫毛部位維持脫毛的皮膚色澤，反觀該實驗組1~3當中各該個體的脫毛部位均開始長出細小的黑色毛髮，從圖1的照片來看，該實驗組2、3以及當中各該個體的脫毛部位可明顯觀察到毛髮生長的成果；在第14天時，該控制組及該實驗組1~3當中各該個體的脫毛部位均生長出黑色毛髮，且從圖1的照片來看，該實驗組1~3當中各該個體的毛髮生長狀況均明顯優於該控制組。Please refer to Figure 1 for the observation results. The control group and the experimental groups 1 to 3 can clearly see the hair loss on the back of each individual on the 1st day; on the 7th day, the hair loss of the individual in the control group The skin color of the depilated area was maintained. On the other hand, the depilated areas of the individuals in the experimental groups 1 to 3 began to grow fine black hairs. Judging from the photos in Figure 1, the experimental groups 2, 3 and the individuals in the experimental groups began to grow fine black hairs. The results of hair growth can be clearly observed in the hair removal area; on the 14th day, black hair grew in the hair removal area of each individual in the control group and experimental groups 1 to 3, and from the photo in Figure 1, the The hair growth status of each individual in experimental groups 1 to 3 was significantly better than that of the control group.
各組個體的脫毛部位皮膚灰度值分析：Analysis of skin gray value of hair removal parts of individuals in each group:
為了進一步探討該控制組及該實驗組1~3當中各該個體的毛髮生長狀況，本試驗選用各組在第14天的各該個體，先對各組個體對應該脫毛部位進行影像擷取，再使用image J軟體分析各組個體的影像灰度值。In order to further explore the hair growth status of the individuals in the control group and the experimental groups 1 to 3, this experiment selected the individuals in each group on the 14th day. First, images were captured of the hair removal parts of the individuals in each group. Then use image J software to analyze the gray value of the images of each group of individuals.
「灰度值」為黑白圖像中的顏色深度，範圍一般從0到255，黑色為0，白色為255，可藉由分析數值的高低來判斷各組個體的毛髮增生情形，其中當分析數值越低且趨近於0時，表示灰度值接近黑色，代表該個體的脫毛部位有明顯的毛髮增生現象；相反地，當分析數值越高時，表示灰度值接近白色，代表該個體的脫毛部位不具毛髮生長現象。"Grayscale value" is the color depth in a black and white image, generally ranging from 0 to 255, with black being 0 and white being 255. The hair growth of each group of individuals can be judged by analyzing the value. When analyzing the value When the value is lower and approaches 0, it means that the gray value is close to black, which means that the individual's hair removal area has obvious hair growth; conversely, when the analysis value is higher, it means that the gray value is close to white, which means that the individual's hair growth is There is no hair growth in the hair removal area.
請參閱圖2及表1的檢測結果，該控制組的皮膚灰度值為119.4577±17.2175，該實驗組1的皮膚灰度值為113.235±38.6152，該實驗組2的皮膚灰度值為78.4813±14.016，該實驗組3的皮膚灰度值為50.8783±5.0039，從檢測結果得知，該實驗組1~3的皮膚灰度值相較於該控制組的皮膚灰度值有下降，表示該實驗組1~3均有效助於毛髮增生的現象，更由實驗組1~3的數據結果來看，該實驗組3的皮膚灰度值為最低，表示該實驗組3的毛髮增生效果最為優異，其次為該實驗組2，由此可判斷該實驗組2、3當中該促進生髮組成物所使用該蘆丁成分的有效量能對該個體的脫毛部位產生優異的毛髮增生效果。Please refer to the test results in Figure 2 and Table 1. The skin gray value of the control group is 119.4577±17.2175, the skin gray value of the experimental group 1 is 113.235±38.6152, and the skin gray value of the experimental group 2 is 78.4813± 14.016, the skin gray value of experimental group 3 is 50.8783±5.0039. From the test results, it is known that the skin gray value of experimental group 1~3 has decreased compared with the skin gray value of the control group, indicating that the experiment Groups 1 to 3 are all effective in promoting hair growth. Judging from the data results of experimental groups 1 to 3, the skin gray value of experimental group 3 is the lowest, which means that experimental group 3 has the best hair growth effect. Next is the experimental group 2. From this, it can be judged that the effective amount of the rutin component used in the hair growth-promoting composition in the experimental groups 2 and 3 can produce an excellent hair growth effect on the hair removal site of the individual.
表1.係為實驗組1~3及控制組分別在14天時所檢測的皮膚灰度值數據表
上述各組皮膚灰度值及標準誤差(Standard Error, SE)的數據結果，係由各組個體進行三個獨立實驗(n=3)的檢測數值，其中「*」表示該實驗組1~3與該控制組的數值比較，p＜0.05。The data results of the above-mentioned skin gray value and standard error (SE) of each group are the detection values of three independent experiments (n=3) conducted by individuals in each group, where "*" indicates the experimental group 1~3 Compared with the value of the control group, p<0.05.
各組個體的毛髮生長之長度測量分析：Measurement and analysis of hair growth length in each group of individuals:
本試驗進一步探討各組個體的毛髮生長的長度，同樣選用各組在第14天的各該個體，試驗過程係對各組個體進行犧牲，從各組個體對應脫毛部位隨機取10根新生毛髮並進行長度量測。This experiment further explores the length of hair growth of individuals in each group. Individuals in each group on the 14th day are also selected. During the test, individuals in each group are sacrificed, and 10 new hairs are randomly taken from the corresponding hair removal parts of each group of individuals and Take length measurements.
請參閱圖3及表2的實驗結果，該控制組的新生毛髮平均長度為0.21±0.0306公分，該實驗組1的新生毛髮平均長度為±0.0569公分，該實驗組2的新生毛髮平均長度為0.5067±0.057公分，該實驗組3的新生毛髮平均長度為0.5467±0.0441公分，從檢測結果得知，該實驗組1~3的新生毛髮平均長度相較於該控制組均呈現增長，更由該實驗組1~3及的數據結果來看，該實驗組2、3的新生毛髮平均長度相較於該實驗組1更具有顯著性的增長，其中又以該實驗組3的新生毛髮平均長度為最長，由此可判斷該實驗組2、3當中該促進生髮組成物所使用該蘆丁成分的有效量能提供較佳的毛髮生長效率。Please refer to the experimental results in Figure 3 and Table 2. The average length of new hair in the control group is 0.21±0.0306 cm, the average length of new hair in experimental group 1 is ±0.0569 cm, and the average length of new hair in experimental group 2 is 0.5067 ±0.057 cm, the average length of new hair in experimental group 3 is 0.5467±0.0441 cm. From the test results, it is known that the average length of new hair in experimental groups 1 to 3 has increased compared with the control group. Moreover, this experiment Judging from the data results of groups 1 to 3 and 3, the average length of new hair in experimental groups 2 and 3 has increased significantly compared with experimental group 1, and the average length of new hair in experimental group 3 is the longest. , it can be judged that the effective amount of the rutin component used in the hair growth-promoting composition in the experimental groups 2 and 3 can provide better hair growth efficiency.
表2.係為實驗組1~3及控制組分別在14天時所檢測的新生毛髮平均長度統計表
上述各組新生毛髮平均長度及標準誤差(Standard Error, SE)的數據結果，係由各組個體進行三個獨立實驗(n=3)的檢測數值，其中「*」表示該實驗組1~3與該控制組的數值比較，p＜0.05。The above data results of the average length and standard error (SE) of new hair in each group are the test values of three independent experiments (n=3) conducted by individuals in each group, where "*" indicates the experimental group 1~3 Compared with the value of the control group, p<0.05.
各組個體的組織切片觀察與皮膚厚度分析：Tissue section observation and skin thickness analysis of individuals in each group:
前述實驗的各組個體犧牲後進行背部皮膚組織的固定 (fixation)、包埋、切片等處理步驟，且於脫蠟後以Hematoxylin & Eosin 染色，有利於評估觀察皮膚及毛囊組織；當皮膚中的毛囊產生生髮現象時，意味著這階段的毛囊處於毛髮生長期，所以在這階段能觀察到狹長型的毛囊形狀以及真皮層的增厚，作為辨別毛髮增生的辨別標準。The individuals in each group of the aforementioned experiments were sacrificed and underwent processing steps such as fixation, embedding, and slicing of the back skin tissue. After dewaxing, they were stained with Hematoxylin & Eosin, which is conducive to the evaluation and observation of skin and hair follicle tissue; when the skin tissue in the skin When hair follicles grow, it means that the hair follicles are in the anagen phase. Therefore, the narrow and long hair follicle shape and the thickening of the dermis can be observed at this stage, which serve as the criteria for identifying hair hyperplasia.
請參閱圖4的影像結果，該控制組的皮膚組織不具明顯的狹長的毛囊形狀，該實驗組1的皮膚組織可直接看出毛囊數目的增加，且在該實驗組2、3更能清楚觀察到相對狹長的毛囊形狀，藉以該實驗組1~3均具備促進毛囊增生的效果。Please refer to the image results in Figure 4. The skin tissue of the control group does not have an obvious long and narrow hair follicle shape. The increase in the number of hair follicles can be directly seen in the skin tissue of the experimental group 1, and it can be more clearly observed in the experimental groups 2 and 3. The relatively long and narrow shape of hair follicles indicates that experimental groups 1 to 3 all have the effect of promoting hair follicle proliferation.
接著，本試驗檢測該控制組及該實驗組1~3的皮膚生長厚度，請參閱圖5及表3的皮膚厚度檢測結果，該控制組的皮膚厚度為742.4537±74.9902微米，而該實驗組1的皮膚厚度為868.3417±31.5609微米，該實驗組2的皮膚厚度為1015.638±29.4934微米，該實驗組3的皮膚厚度為1011.933±29.4934微米，從檢測結果得知，該實驗組1~3相較於該控制組確實能有效促進皮膚厚度增生的現象，其中以該實驗組2、3的皮膚厚度增長最為顯著。Next, this test detects the skin growth thickness of the control group and the experimental group 1 to 3. Please refer to the skin thickness detection results in Figure 5 and Table 3. The skin thickness of the control group is 742.4537±74.9902 microns, while the skin thickness of the experimental group 1 The skin thickness of experimental group 2 is 868.3417±31.5609 microns, the skin thickness of experimental group 2 is 1015.638±29.4934 microns, and the skin thickness of experimental group 3 is 1011.933±29.4934 microns. From the test results, it is known that compared with experimental groups 1~3 The control group can indeed effectively promote the proliferation of skin thickness, among which the skin thickness growth of experimental groups 2 and 3 is the most significant.
表3.係為實驗組1~3及控制組在14天時所檢測的皮膚厚度數據表
上述各組皮膚厚度及標準誤差(Standard Error, SE)的數據結果，係由各組個體進行三個獨立實驗(n=3)的檢測數值，其中「*」表示該實驗組1~3與該控制組的數值比較，p＜0.05。The above-mentioned data results of skin thickness and standard error (SE) of each group are the detection values of three independent experiments (n=3) conducted by individuals in each group, where "*" indicates that the experimental group 1~3 and the Numerical comparison of the control group, p<0.05.
分析各組個體皮膚組織當中β-連環蛋白（β-catenin）表現：Analyze the expression of β-catenin in skin tissues of individuals in each group:
將各組犧牲後個體進行背部皮膚組織的均質化、蛋白質裂解萃取、蛋白質電泳、SDS-Page展開蛋白及蛋白轉漬處理至PVDF膜等處理步驟，接著使用甲醇復水及使用脫脂奶粉進行蛋白阻攔後，以β-連環蛋白（β-catenin）一級抗體（Santa Cruz, sc-7963）與小鼠二級抗體進行轉染，並使用enhanced luminal reagent進行呈色，利用迷你型冷光成像分析系統及其軟體Evolution Capt 進行影像擷取與分析。當皮膚毛囊活化產生相關生髮現象時，β-連環蛋白（β-catenin）的活化能提高毛囊細胞的活性以調控生髮現象。The sacrificed individuals in each group were subjected to processing steps such as homogenization of back skin tissue, protein lysis extraction, protein electrophoresis, SDS-Page unfolding of proteins, and protein transfer to PVDF membrane, followed by rehydration with methanol and protein blocking with skim milk powder. After that, β-catenin primary antibody (Santa Cruz, sc-7963) and mouse secondary antibody were used for transfection, and enhanced luminal reagent was used for color development, using a mini luminescence imaging analysis system and its The software Evolution Capt performs image capture and analysis. When skin hair follicles are activated to produce related hair growth phenomena, the activation of β-catenin can increase the activity of hair follicle cells to regulate hair growth phenomena.
請參閱圖6及表4的試驗結果，該控制組的β-連環蛋白（β-catenin）表現量為100%，而該實驗組1的β-連環蛋白（β-catenin）相對於該控制組的表現量為246.595%±25.5353，該實驗組2的β-連環蛋白（β-catenin）相對於該控制組的表現量為263.251%±48.0316，該實驗組3的β-連環蛋白（β-catenin）相對於該控制組的表現量為293.0717%±22.5779，由數據結果可清楚得知，該實驗組1~3當中該促進生髮組成物之蘆丁成分有效量能讓該β-連環蛋白的表現量提升至少200%，在一較佳實施例中，該β-連環蛋白的表現量提升200%~350%，藉以該實驗組1~3相較於該控制組確實能顯著地活化β-連環蛋白（β-catenin）的表現，其中以該實驗組2、3的β-連環蛋白（β-catenin）表現量更為顯著，在本實施例中，該實驗組2、3當中該促進生髮組成物之蘆丁成分有效量能讓β-連環蛋白（β-catenin）表現量提升215%~315%，在一較佳實施例中，該實驗組2、3當中該促進生髮組成物之蘆丁成分有效量能讓β-連環蛋白（β-catenin）表現量提升260%~300%；藉此，該實驗組1~3當中蘆丁成分的有效量均能有效活化β-連環蛋白（β-catenin）的表現，在一較佳實施例中，該實驗組2、3當中β-連環蛋白（β-catenin）活化表現為較佳，其中又以該實驗組3當中β-連環蛋白（β-catenin）活化表現為最佳，且該蘆丁成分經一系列實驗證實能有助於增進毛髮生長的效果。Please refer to the test results in Figure 6 and Table 4. The expression amount of β-catenin in the control group is 100%, while the β-catenin in the experimental group 1 is lower than that in the control group. The expression amount of β-catenin (β-catenin) in experimental group 2 relative to the control group was 263.251% ± 48.0316, and the β-catenin (β-catenin) in experimental group 3 was 246.595% ± 25.5353. ) relative to the expression amount of the control group is 293.0717% ± 22.5779. It can be clearly seen from the data results that the effective amount of the rutin component of the hair growth-promoting composition in the experimental groups 1 to 3 can make the expression of β-catenin The amount of β-catenin is increased by at least 200%. In a preferred embodiment, the expression amount of β-catenin is increased by 200% to 350%, so that the experimental groups 1 to 3 can indeed significantly activate β-catenin compared with the control group. The expression of protein (β-catenin), among which the expression amount of β-catenin (β-catenin) in experimental groups 2 and 3 is more significant. In this example, the hair growth-promoting composition in experimental groups 2 and 3 An effective amount of the rutin component of the substance can increase the expression of β-catenin by 215% to 315%. In a preferred embodiment, the rutin of the hair growth-promoting composition in the experimental groups 2 and 3 The effective dose of the ingredient can increase the expression of β-catenin by 260% to 300%. Therefore, the effective dose of rutin in experimental groups 1 to 3 can effectively activate β-catenin (β-catenin). catenin) performance, in a preferred embodiment, the activation performance of β-catenin (β-catenin) in the experimental groups 2 and 3 is better, and the β-catenin (β-catenin) in the experimental group 3 has better activation performance. catenin) has the best activation performance, and this rutin ingredient has been confirmed by a series of experiments to help promote hair growth.
表4.係為實驗組1~3在14天時所檢測β-catenin 相對控制組的表現量數據表
上述各組β-連環蛋白相對表現量及標準誤差(Standard Error, SE)的數據結果，係由各組個體進行三個獨立實驗(n=3)的檢測數值，其中「*」表示該實驗組1~3與該控制組的數值比較，p＜0.05。The data results of the relative expression amount and standard error (SE) of β-catenin in each group above are the detection values of three independent experiments (n=3) conducted by individuals in each group, where "*" indicates the experimental group 1~3 compared with the value of the control group, p<0.05.
歸納上述，該促進生髮組成物當中該蘆丁成分為天然植物的萃取物，在皮膚使用上無副作用產生，且經過一系列動物試驗顯示，該個體在持續外用施予該促進生髮組成物7~14天時，該蘆丁成分能夠顯著活化該個體的脫毛部位當中皮膚組織的β-連環蛋白（β-catenin）表現，使得該個體的脫毛部位能明顯看出皮膚毛髮增生，其中以該實驗組2、3當中該蘆丁成分使用所述有效量對應β-連環蛋白（β-catenin）活化表現為較佳，藉以有效提供優異促進毛髮生長及預防脫髮的效果。To sum up the above, the rutin component in the hair growth-promoting composition is a natural plant extract and has no side effects when used on the skin. A series of animal tests have shown that the individual continued to apply the hair growth-promoting composition 7~ At 14 days, the rutin component could significantly activate the expression of β-catenin in the skin tissue of the individual's hair removal area, so that the skin and hair proliferation could be clearly seen in the individual's hair removal area. Among them, the experimental group Among 2 and 3, the rutin component performs better in activating β-catenin when used in the stated effective amount, thus effectively providing excellent effects in promoting hair growth and preventing hair loss.
以上所述僅為本發明較佳可行實施例而已，並非用以限制本發明，本發明之技術領域的通常知識者，於參酌以上教示後，當能對上述實施例的內容做適當的修改或根據不同的實驗方法，達成本發明所主張的技術功效。The above descriptions are only preferred preferred embodiments of the present invention and are not intended to limit the present invention. A person with ordinary skill in the technical field of the present invention can make appropriate modifications to the contents of the above embodiments after referring to the above teachings. According to different experimental methods, the technical effects advocated by the present invention are achieved.
另外，以上β-連環蛋白表現量提升的幅度可能因為實驗之個體差異而有所不同，因此，本發明之促進生髮組成物的效果並不以上述實驗所示出的提升量為限，舉凡應用本發明說明書及申請專利範圍所為之等效變化，理應包含在本發明之專利範圍內。In addition, the above-mentioned increase in β-catenin expression may vary due to individual differences in experiments. Therefore, the effect of the hair growth-promoting composition of the present invention is not limited to the increase shown in the above-mentioned experiments. It applies to all applications. Equivalent changes made in the description of the invention and the patent application scope should be included in the patent scope of the invention.
［本發明］ 無 [Invention] without
圖1係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在不同時間觀察之影像對照圖。 圖2係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在14天時所檢測的皮膚灰度值數據圖。 圖3係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在14天時所檢測的新生毛髮平均長度數據圖。 圖4係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在14天時所觀察的皮膚組織切片圖。 圖5係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在14天時所檢測的皮膚厚度數據圖。 圖6係為本發明一較佳實施例之蘆丁用於製備促進生髮組成物的用途當中各實驗組在14天時所檢測的β-catenin相對表現量數據圖。 Figure 1 is a comparison of images observed by each experimental group at different times when rutin is used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention. Figure 2 is a graph showing the skin gray value data of each experimental group measured at 14 days when rutin was used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention. Figure 3 is a graph showing the average length of new hair detected in each experimental group at 14 days when rutin is used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention. Figure 4 is a diagram of skin tissue sections observed in each experimental group at 14 days when rutin was used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention. Figure 5 is a graph showing the skin thickness data of each experimental group measured at 14 days when rutin was used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention. Figure 6 is a data chart showing the relative expression amount of β-catenin detected in each experimental group at 14 days when rutin was used to prepare a composition for promoting hair growth according to a preferred embodiment of the present invention.
Claims (5)
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0377809A (en) * | 1989-08-21 | 1991-04-03 | Arusoa Sogo Kenkyusho:Kk | Hair tonic |
| JPH04275210A (en) * | 1991-03-04 | 1992-09-30 | Sadao Tsukada | Hair tonic and growing agent |
| JPH0517321A (en) * | 1991-07-12 | 1993-01-26 | Goro Kawaguchi | Hair grower |
| CN112120953A (en) * | 2019-06-24 | 2020-12-25 | 珀莱雅化妆品股份有限公司 | Preparation method of freeze-dried liposome containing hair-nourishing composition |
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2022
- 2022-11-02 TW TW111141869A patent/TWI827334B/en active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0377809A (en) * | 1989-08-21 | 1991-04-03 | Arusoa Sogo Kenkyusho:Kk | Hair tonic |
| JPH04275210A (en) * | 1991-03-04 | 1992-09-30 | Sadao Tsukada | Hair tonic and growing agent |
| JPH0517321A (en) * | 1991-07-12 | 1993-01-26 | Goro Kawaguchi | Hair grower |
| CN112120953A (en) * | 2019-06-24 | 2020-12-25 | 珀莱雅化妆品股份有限公司 | Preparation method of freeze-dried liposome containing hair-nourishing composition |
Non-Patent Citations (1)
| Title |
|---|
| 期刊 J Huelsken, et al. " beta-Catenin controls hair follicle morphogenesis and stem cell differentiation in the skin." Cell. ;105(4) ;2001 May 18 :533-545. * |
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