TW202228741A - 具有生髮、養髮效果之乳酸菌 - Google Patents
具有生髮、養髮效果之乳酸菌 Download PDFInfo
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- TW202228741A TW202228741A TW110136189A TW110136189A TW202228741A TW 202228741 A TW202228741 A TW 202228741A TW 110136189 A TW110136189 A TW 110136189A TW 110136189 A TW110136189 A TW 110136189A TW 202228741 A TW202228741 A TW 202228741A
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- Prior art keywords
- lactic acid
- acid bacteria
- hair
- present
- dermal papilla
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Abstract
本發明利用毛乳頭細胞之活化及角質細胞增殖因子對獲得生髮、養髮、護髮效果而言必不可少之觀點，探索具有該活化能力及該增殖因子產生能力之乳酸菌。
本發明利用具有毛乳頭細胞活化能力或角質細胞增殖因子產生促進能力之乳酸桿菌屬乳酸菌、其培養液或醱酵產物。又，該乳酸菌較佳為具有毛乳頭細胞活化能力及角質細胞增殖因子產生促進能力兩者。進而，作為本案發明之乳酸菌之例示，可列舉胚芽乳酸桿菌N793株(NITE BP-03233)。
Description
本發明係一種具有較高之毛乳頭細胞活化能力及角質細胞增殖因子產生促進能力之源自玉米之乳酸桿菌屬胚芽乳酸桿菌種之乳酸菌之菌株、以及含有該乳酸菌、其培養液或醱酵產物之處理物之飲料、食品、化妝品、準醫藥品及醫藥品。
毛髮稀少係指因年齡增長、壓力、睡眠不足、遺傳等各種原因而導致髮根活性下降，毛髮週期變短。毛髮週期係指毛髮重複生長期、退行期、休止期3個階段之週期(非專利文獻1)。毛髮週期中最長之時期為生長期，通常為3年〜6年左右。但是，因上述毛髮週期變短之原因，導致生長期縮短，毛髮稀少。
生長期係藉由角質細胞增殖因子、類胰島素生長因子1、血管內皮增殖因子等生長因子來維持。該等生長因子係由存在於髮根深處之毛乳頭細胞產生。已知，自毛乳頭細胞分泌之角質細胞增殖因子係直接作用於成為毛髮來源之毛母細胞，促進毛母細胞之增殖及分裂，故認為其係最直接作用於毛髮之生長因子。根據以上情況，毛乳頭細胞係生髮所必需之細胞，若可促進毛乳頭細胞之活化及角質細胞增殖因子產生量之增加，則有助於生髮(非專利文獻2)。
另一方面，於專利文獻1中揭示有一種以生髮及養髮為目的而利用植物提取物作為促進角質細胞增殖之促進劑的方法。
[先前技術文獻]
[非專利文獻]
[非專利文獻1]野野村美宗著，「化妝品科學」，第1版，Fragrance Journal公司，2020年3月，p.74-78
[非專利文獻2]M. Iino et al., 「Adenosine Stimulates Fibroblast Growth Factor-7 Gene Expression Via Adenosine A2b Receptor Signaling in Dermal Papilla Cells」, Vol.127, Journal of Investigative Dermatology, June 2007, p.1318-1325
[專利文獻]
[專利文獻1]日本專利特開2009-242324
又，亦設想該先前申請案公開公報中所揭示之利用植物提取物之方法以外的方法。
(發明所欲解決之問題)
因此，本發明人等著眼於乳酸菌，利用毛乳頭細胞之活化及角質細胞增殖因子對獲得生髮、養髮、護髮效果而言必不可少之觀點，對數種乳酸菌進行篩選，結果發現存在具有較高之毛乳頭細胞活化能力及角質細胞增殖因子產生促進能力之乳酸菌，從而完成了本發明。
(解決問題之技術手段)
亦即，本案發明之第1態樣係一種乳酸桿菌屬乳酸菌、其培養液或醱酵產物，其等具有毛乳頭細胞活化能力或角質細胞增殖因子產生促進能力之至少一者。本案發明之第2態樣係如本案發明之第1態樣所記載之乳酸菌、其培養液或醱酵產物，其等具有毛乳頭細胞活化能力及角質細胞增殖因子產生促進能力兩者。本案發明之第3態樣係如本案發明之第1或2態樣所記載之乳酸菌，其中，上述乳酸桿菌屬乳酸菌為胚芽乳酸桿菌N793株(NITE BP-03233)。
本案申請人亦追求一種包含該乳酸菌、其培養液或醱酵產物之飲食品。亦即，本案發明之第4態樣係一種飲食品，其含有本案發明之第1至3態樣中任一項所記載之乳酸菌、其培養液或醱酵產物。
進而，本案申請人亦追求一種包含該乳酸菌、其培養液或醱酵產物之化妝品、準醫藥品、醫藥品。亦即，本案發明之第5態樣係一種化妝品、準醫藥品或醫藥品，其等含有本案發明之第1至3態樣中任一項所記載之乳酸菌、其培養液或醱酵產物。
(對照先前技術之功效)
本發明之乳酸菌、其培養液或醱酵產物係具有毛乳頭細胞活化能力或角質細胞增殖因子產生促進能力之至少一者。
以下，對本發明詳細地進行說明。
1.胚芽乳酸桿菌N793株(NITE BP-03233)
本發明之乳酸菌為胚芽乳酸桿菌(Lactobacillus plantarum)。尤其是胚芽乳酸桿菌屬乳酸菌中之胚芽乳酸桿菌N793株(NITE BP-03233)。本發明中所謂之記號N793係日清食品控股股份有限公司自行賦予菌株之編號，本胚芽乳酸桿菌N793株係本發明人首次分離所得者。
再者，更詳細而言，本胚芽乳酸桿菌N793株被鑑定為胚芽乳酸桿菌胚芽亞種(Lactobacillus plantarum subsp. plantarum)屬乳酸菌。
本發明之胚芽乳酸桿菌N793株係於2020年6月18日以NITE BP-03233寄存於獨立行政法人製品評價技術基盤機構專利微生物寄存中心。本發明之胚芽乳酸桿菌N793株之菌學性質係如以下表1及表2所示。本菌學性質係基於伯傑氏系統細菌學手冊(Bergey's manual of systematic bacteriology)Vol.2 (1986)中記載之方法。表1係表示與本菌株相關之形狀等，表2係表示藉由API 50CH及API CHL(Biomerieux公司製造)對糖合成代謝性進行試驗所得之結果。表2中，「+」表示陽性，「－」表示陰性。
[表1]
| 試驗項目 | 胚芽乳酸桿菌N793株 | |
| 培養溫度 | 30℃ | |
| 細胞形態 | 桿菌 (0.6～0.7×2.0～3.0 μm) | |
| 革蘭氏染色性 | + | |
| 有無芽孢 | － | |
| 運動性 | － | |
| 菌落形態 | 培養基 | MRS瓊脂培養基 |
| 培養時間 | 24小時 | |
| 直徑 | 2～3 mm | |
| 色調 | 乳白色 | |
| 形狀 | 圓形 | |
| 隆起狀態 | 透鏡狀 | |
| 周緣 | 全緣 | |
| 表面之形狀等 | 平滑 | |
| 透明度 | 不透明 | |
| 黏稠度 | 黃油狀 | |
| 生長溫度試驗 | 37℃ | + |
| 45℃ | + | |
| 觸酶反應 | － | |
| 氧化酶反應 | － | |
| 自葡萄糖產生酸/氣體 | +/－ | |
| O/F測試(氧化/醱酵) | +/－ | |
| +：陽性、－：陰性 |
[表2]
| 糖基質 | 結果 | 糖基質 | 結果 | ||
| 0 | 對照組 | － | 25 | 馬栗樹皮苷檸檬酸鐵 | + |
| 1 | 甘油 | － | 26 | 柳苷 | + |
| 2 | 赤藻糖醇 | － | 27 | D-纖維二糖 | + |
| 3 | D-阿拉伯糖 | － | 28 | D-麥芽糖 | + |
| 4 | L-阿拉伯糖 | + | 29 | D-乳糖 | + |
| 5 | D-核糖 | + | 30 | D-蜜二糖 | + |
| 6 | D-木糖 | － | 31 | D-蔗糖 | + |
| 7 | L-木糖 | － | 32 | D-海藻糖 | + |
| 8 | D-福壽草醇 | － | 33 | 菊糖 | － |
| 9 | 甲基-βD-吡喃木糖苷 | － | 34 | D-松三糖 | + |
| 10 | D-半乳糖 | + | 35 | D-棉子糖 | + |
| 11 | D-葡萄糖 | + | 36 | 澱粉 | － |
| 12 | D-果糖 | + | 37 | 肝糖 | － |
| 13 | D-甘露糖 | + | 38 | 木糖醇 | － |
| 14 | L-花椒糖 | － | 39 | 龍膽二糖 | + |
| 15 | L-鼠李糖 | － | 40 | D-松二糖 | + |
| 16 | 半乳糖醇 | － | 41 | D-來蘇糖 | － |
| 17 | 肌醇 | － | 42 | D-塔格糖 | － |
| 18 | D-甘露醇 | + | 43 | D-岩藻糖 | － |
| 19 | D-山梨糖醇 | + | 44 | L-岩藻糖 | － |
| 20 | 甲基-αD-吡喃甘露糖苷 | + | 45 | D-阿拉伯糖醇 | － |
| 21 | 甲基-αD-吡喃葡萄糖苷 | － | 46 | L-阿拉伯糖醇 | － |
| 22 | N-乙醯葡糖胺 | + | 47 | 葡萄糖酸鹽 | + |
| 23 | 苦杏仁苷 | + | 48 | 2-酮葡萄糖酸鹽 | － |
| 24 | 熊果苷 | + | 49 | 5-酮葡萄糖酸鹽 | － |
[表3]
| 朊蛋白腖No.3 | 10.00 g |
| 牛肉萃取物 | 10.00 g |
| 酵母萃取物 | 5.00 g |
| D-葡萄糖 | 20.00 g |
| 聚山梨醇酯80 | 1.00 g |
| 檸檬酸銨 | 2.00 g |
| 乙酸鈉 | 5.00 g |
| 硫酸錳 | 0.10 g |
| 硫酸鎂 | 0.05 g |
| 磷酸二鉀 | 2.00 g |
| 蒸餾水 | 1000 mL |
2.毛乳頭細胞活化試驗
如下述實驗例所示，本發明之胚芽乳酸桿菌N793株係具有較高之毛乳頭細胞活化能力。毛乳頭細胞活化能力之確認係藉由以下試驗方法來進行。
＜用於毛乳頭細胞活化能力評價之受檢體(乳酸菌粉末懸浮液)之製備＞
用於毛乳頭細胞活化能力評價之受檢體(乳酸菌粉末懸浮液)係將乳酸菌於表3所示之MRS培養基(Becton Dickinson公司製造)中於37℃下培養24小時。繼而，使用離心分離機收集經增殖之菌體。用殺菌水將收集之菌體洗淨，藉由離心分離機收集菌體。將洗淨及收集菌體反覆進行3次。於95℃下加熱殺菌15分鐘後，再次藉由離心分離機收集菌體。使用冷凍乾燥機將收集之菌體進行冷凍乾燥，獲得乾燥菌體粉末。使乾燥菌體粉末以1 mg/mL懸浮於磷酸鹽緩衝液(PBS，Phosphate Buffered Saline)(PBS(－) pH 7.4 Gibco公司製造)中，製成乳酸菌粉末懸浮液。
＜毛乳頭細胞活化能力評價＞
對上述獲得之受檢體之毛乳頭細胞活化作用進行評價。亦即，將人毛乳頭細胞以1.2×10
4cells/well之密度接種於96孔板，於CO
2培養箱(5%CO
2、37℃)中培養6小時後，添加包含各受檢體(最終濃度1%)之100 μL之D-MEM培養基(FBS4%)。添加後，於CO
2培養箱(5%CO
2、37℃)中培養72小時後，藉由吸光光度法測定培養液中之毛乳頭細胞數。測定時使用細胞計數套組8(同仁化學研究所製造)。再者，使用PBS(PBS(－) pH 7.4 Gibco公司製造)作為對照組。
3.角質細胞增殖因子產生促進試驗
如下述實驗例所示，本發明之胚芽乳酸桿菌N793株係具有較高之角質細胞增殖因子產生促進能力。角質細胞增殖因子產生促進能力之確認係藉由以下試驗方法來進行。
＜角質細胞增殖因子產生促進能力評價＞
對與毛乳頭細胞活化試驗相同之受檢體評價角質細胞增殖因子產生促進作用。將人毛乳頭細胞以1.0×10
4cells/well之密度接種於48孔板，於CO
2培養箱(5%CO
2、37℃)中培養24小時後，添加包含各受檢體群(最終濃度10%)之300 μL之毛乳頭細胞培養基(PromoCell公司製造)。添加後，於CO
2培養箱(5%CO
2、37℃)中培養24小時後，藉由酵素結合免疫吸附分析(ELISA，enzyme linked immunosorbent assay)法測定培養上清液中之角質細胞增殖因子。測定時係使用人KGF/FGF-7 DuoSet ELISA(R&D Systems公司製造)。再者，使用PBS(PBS(－) pH 7.4 Gibco公司製造)作為對照組。
4.菌體、其培養液或醱酵產物
本發明之乳酸菌可以各種形態利用。例如，若為菌體本身，則可利用該菌體之冷凍乾燥型或對該菌體進行酵素處理所得者等。
進而，除了菌體本身以外，亦可利用本發明之乳酸菌之培養液。具體而言，菌體、菌體及培養液、培養上清液等均可利用。又，當然，可對上述菌體等視需要實施離心分離或者蛋白酶或脂肪酶等酵素處理。
進而，亦可為利用菌體、其培養液以及該乳酸菌之醱酵產物。即，亦可利用以糖類等作為營養源並藉由該乳酸菌進行醱酵所獲得之醱酵產物。具體而言，作為以乳等作為原料之醱酵產物，可列舉酸酪乳或乳酪。
5.飲食品
本發明之乳酸菌、其培養液或醱酵產物可包含於飲食品中來使用。例如考慮添加有乳酸菌之醱酵乳及添加有乳酸菌之乳酸菌飲料。又，作為醱酵產物，可列舉酸酪乳或乳酪等。於目前之與乳及乳製品之成分標準等相關之政府機關指令中，作為成分標準，乳酸菌數並無特別規定，若為醱酵乳(無脂乳固形份為8.0%以上者)或乳酸菌飲料(無脂乳固形份為3.0%以上者)，則較佳為1.0×10
7cfu/ml以上，若為乳酸菌飲料(無脂乳固形份未滿3.0%者)，則較佳為1.0×10
6cfu/ml以上，藉由於乳等醱酵液中增殖，或以最終製品之形態增殖，可實現上述菌數。又，除了添加有乳酸菌之醱酵乳及添加有乳酸菌之乳酸菌飲料以外，亦可用於黃油等乳製品、蛋黃醬等蛋加工品、乳酪餅等甜點麵包類等。又，亦可適用於泡麵或曲奇等加工食品。除了上述以外，本發明之食品亦可為將上述乳酸菌、其培養液或醱酵產物與視需要之適當載體及添加劑一起添加並進行製劑化而成之形態(例如粉末、顆粒、膠囊、錠劑等)。又，包含本發明之乳酸菌、其培養液或醱酵產物之飲食品只要含有本發明之菌體、其培養液或醱酵產物作為有效成分即可，當然，除了本發明之乳酸菌、其培養液或醱酵產物以外，亦可含有其他成分(例如糖類、蛋白質、胺基酸、脂質、水分、維生素、礦物質等)。
本發明之乳酸菌、其培養液或醱酵產物除了包含於一般之飲料或食品中有用以外，包含於特定保健用食品、營養輔助食品等中亦有用。
6.化妝品、準醫藥品、醫藥品
本發明之乳酸菌、其培養液或醱酵產物除了可包含於食品中來使用以外，亦可包含於化妝品、準醫藥品、醫藥品中來使用。具體而言，可列舉：護髮劑、養髮劑、生髮劑、或利用該等之洗髮精、潤絲精、生髮水、養髮露等。
亦即，可藉由濃縮或分離等將本發明之乳酸菌、其培養液或醱酵產物進行萃取物化或粉末化而製備後，用作以該成分作為有效成分之化妝品、準醫藥品、醫藥品。例如，可提供將經萃取物化者裝入瓶等中者，或亦使用賦形劑等將醱酵液乾燥粉末以顆粒或膠囊化、打錠而成錠劑之形態提供。
又，作為化妝品、準醫藥品、醫藥品，只要含有本發明之菌體、其培養液或醱酵產物作為有效成分即可，當然，除了該成分以外，亦可含有其他成分(例如糖類、蛋白質、胺基酸、脂質、水分、維生素、礦物質等)。
進而，本發明之乳酸菌、其培養液或醱酵產物亦可應用於青貯料、動物用餌料、植物液體肥料等動物飼料、植物肥料領域。
(產業上之可利用性)
本發明之乳酸菌(胚芽乳酸桿菌N793株)、其培養液或醱酵產物係具有較高之毛乳頭細胞活化能力或角質細胞增殖因子產生促進能力之至少一者。
[實施例]
以下，表示本發明之實施例，但本發明並不限定於以下實施例。
＜試驗例1＞毛乳頭細胞活化能力評價
對本發明之胚芽乳酸桿菌N793株及本公司持有之3株胚芽乳酸桿菌比較株(N808、N814、N828)實施毛乳頭細胞活化能力評價。
毛乳頭細胞活化能力評價係按如下順序進行。對於本發明之菌株及比較菌株，分別於表3所示之MRS培養基(Becton Dickinson公司製造)中於CO
2培養箱(5%CO
2、37℃)中培養24小時。繼而，使用離心分離機收集經增殖之菌體。用殺菌水將收集之菌體洗淨，藉由離心分離機進行收集菌體。將洗淨及收集菌體反覆進行3次。其後，使用冷凍乾燥機將收集之菌體進行冷凍乾燥，獲得乾燥菌體粉末。使乾燥菌體粉末以1 mg/mL懸浮於PBS(PBS(－) pH 7.4 Gibco公司製造)中，製成乳酸菌粉末懸浮液。
另外，將人毛乳頭細胞以1.2×10
4cells/well之密度接種於96孔板，於CO
2培養箱(5%CO
2、37℃)中培養6小時後，添加包含各受檢體群(最終濃度1%)之100 μL之D-MEM培養基(FBS4%)。添加後，於CO
2培養箱(5%CO
2、37℃)中培養72小時後，藉由吸光光度法測定培養液中之毛乳頭細胞數。測定時係使用細胞計數套組8(同仁化學研究所製造)。再者，使用PBS(PBS(－) pH 7.4 Gibco公司製造)作為對照組。
表4及圖1中表示將對照組設為1時之各菌株之毛乳頭細胞活化率。
[表4]
| 菌株名稱 | 毛乳頭細胞活化率(vs對照組) |
| 對照組(不添加乳酸菌) | 1.000 |
| 胚芽乳酸桿菌N808 | 1.068 |
| 胚芽乳酸桿菌N814 | 1.062 |
| 胚芽乳酸桿菌N828 | 1.021 |
| 胚芽乳酸桿菌N793 | 1.706 |
根據表4及圖1亦可知，本發明之乳酸菌(胚芽乳酸桿菌N793株)之毛乳頭細胞活化能力明顯較高，與本公司持有之其他乳酸菌相比亦具有較高之毛乳頭細胞活化能力。
＜試驗例2＞角質細胞增殖因子產生促進能力評價
對本發明之胚芽乳酸桿菌N793株、標準株(JCM1149)及本公司持有之3株胚芽乳酸桿菌比較株(N800、N807、N813)實施角質細胞增殖因子產生促進能力評價。
對與毛乳頭細胞活化試驗相同之受檢體評價角質細胞增殖因子產生促進作用。將人毛乳頭細胞以1.0×10
4cells/well之濃度接種於48孔板，於CO
2培養箱(5%CO
2、37℃)中培養24小時後，添加包含各受檢體群(最終濃度10%)之300 μL之毛乳頭細胞培養基(PromoCell公司製造)。添加後，於CO
2培養箱(5%CO
2、37℃)中培養24小時後，藉由ELISA法測定角質細胞增殖因子產生量。測定時係使用人KGF/FGF-7 DuoSet ELISA(R&D Systems公司製造)。再者，使用PBS(PBS(－) pH 7.4 Gibco公司製造)作為對照組。
將各菌株之角質細胞增殖因子產生量示於表5及圖2。
[表5]
| 菌株名稱 | 角質細胞增殖因子產生量(pg/mL) |
| 對照組(不添加乳酸菌) | 28.08 |
| 胚芽乳酸桿菌JCM1149 | 28.29 |
| 胚芽乳酸桿菌N800 | 29.08 |
| 胚芽乳酸桿菌N807 | 31.11 |
| 胚芽乳酸桿菌N813 | 29.53 |
| 胚芽乳酸桿菌N793 | 40.26 |
確認到本發明之乳酸菌(胚芽乳酸桿菌N793株)之角質細胞增殖因子產生促進能力明顯較高，具有與本公司持有之其他乳酸菌相比更高之角質細胞增殖因子產生促進能力。
圖1係將本發明之菌株與比較菌株之毛乳頭細胞活化能力進行比較之圖。
圖2係將本發明之菌株與比較菌株之角質細胞增殖因子產生促進能力進行比較之圖。
寄存國家 JP日本
寄存機構 獨立行政法人製品評價技術基盤機構特許微生物寄託中心(NPMD)
寄存日期 2020/06/18
寄存號碼 BP-03233
Claims (5)
- 一種乳酸桿菌屬乳酸菌、其培養液或醱酵產物，其等具有毛乳頭細胞活化能力或角質細胞增殖因子產生促進能力之至少一者。
- 如請求項1之乳酸菌、其培養液或醱酵產物，其等具有毛乳頭細胞活化能力及角質細胞增殖因子產生促進能力兩者。
- 如請求項1或2之乳酸菌、其培養液或醱酵產物，其中，上述乳酸桿菌屬乳酸菌為胚芽乳酸桿菌N793株(NITE BP-03233)。
- 一種飲食品，其含有請求項1至3中任一項之乳酸菌、其培養液或醱酵產物。
- 一種化妝品、準醫藥品或醫藥品，其含有請求項1至3中任一項之乳酸菌、其培養液或醱酵產物。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2020-176970 | 2020-10-21 | ||
| JP2020176970A JP2022068040A (ja) | 2020-10-21 | 2020-10-21 | 発毛・育毛効果を有する乳酸菌 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| TW202228741A true TW202228741A (zh) | 2022-08-01 |
Family
ID=81456020
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| TW110136189A TW202228741A (zh) | 2020-10-21 | 2021-09-29 | 具有生髮、養髮效果之乳酸菌 |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP4234680A4 (zh) |
| JP (1) | JP2022068040A (zh) |
| CN (1) | CN116171114A (zh) |
| TW (1) | TW202228741A (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2023158297A (ja) | 2022-04-18 | 2023-10-30 | 日東電工株式会社 | ホワイトボードフィルムシステム、ホワイトボードシステムおよび覗き見防止システム |
| CN115501165B (zh) * | 2022-09-22 | 2024-06-11 | 深圳市兰亭科技股份有限公司 | 一种用于头皮护理的组合物及其制备方法与应用 |
-
2020
- 2020-10-21 JP JP2020176970A patent/JP2022068040A/ja active Pending
-
2021
- 2021-09-29 TW TW110136189A patent/TW202228741A/zh unknown
- 2021-10-20 EP EP21882879.6A patent/EP4234680A4/en active Pending
- 2021-10-20 CN CN202180063769.6A patent/CN116171114A/zh active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| EP4234680A4 (en) | 2024-08-21 |
| JP2022068040A (ja) | 2022-05-09 |
| CN116171114A (zh) | 2023-05-26 |
| EP4234680A1 (en) | 2023-08-30 |
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