以下，列舉本發明之實施形態，對本發明進一步詳細地進行說明，但本發明並不限定於該等。以下，於本發明中，將包含鳶尾黃素及鳶尾黃素糖苷之組合物、及包含該等之減少脂肪用組合物、抗肥胖用組合物、促進組織分化用組合物、促進脂肪分解用組合物、用於提高角質層水分量、提高皮膚彈力或抑制皮膚水分蒸散之組合物、改善肝功能用組合物統稱為「本發明之組合物」。 (鳶尾黃素) 本發明中所使用之鳶尾黃素係由分子式C16
H12
O6
表示，亦有稱為5,7-二羥基-3-(4-羥基苯基)-6-甲氧基-4H-1-苯并吡喃-4-酮(5,7-Dihydroxy-3-(4-hydroxyphenyl)-6-methoxy-4H-1-benzopyran-4-one)或者6-
甲氧基-5,7-二羥基-3-(4-羥基苯基)-4H-1-苯并吡喃-4-酮(6-
Methoxy-5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one)之情形。以下亦將鳶尾黃素記載為TGE。鳶尾黃素之化學式如下所示。 [化1](鳶尾黃素糖苷) 作為鳶尾黃素糖苷(以下亦稱為TGE糖苷)，係將上述TGE中1處或2處之氫取代為糖而成者，例如可列舉：鍵結選自葡萄糖、木糖、山梨糖、半乳糖、洋芹糖、鼠李糖之1種或2種以上之糖而成者。就提高促進組織分化、促進脂肪分解、改善肝功能、改善膚質等本發明之效果之觀點而言，較佳為於TGE上鍵結選自葡萄糖、木糖之1種或2種以上之糖而成者。於糖苷中，該等糖通常與TGE之4位及/或7位之羥基鍵結，就更確實地獲得上述效果之方面或糖苷之獲取容易性之方面而言，較佳為與7位之羥基鍵結者。TGE糖苷中之糖之數量(亦稱為糖之鍵結數)例如可列舉1個以上，就提高上述效果之觀點而言，糖之鍵結數較佳為1個〜3個，更佳為1〜2個。此處所謂糖之數量，係指單糖單元之數量。作為TGE糖苷，可僅使用1種，但較佳為組合2種以上。TGE及TGE糖苷於本發明之組合物中，可為有機合成品，亦可從植物等萃取。TGE及TGE糖苷可為粉末狀等固體狀，亦可為溶解於水或有機溶劑中之狀態。TGE及TGE糖苷亦可僅以該等之混合物之形式存在，亦可為溶解、分散或混合至其他物質中之狀態。 作為於TGE上鍵結1個糖而成之糖苷，例如可列舉：鳶尾苷(Tectoridin)、鳶尾苷4'-葡糖苷。又，作為於TGE上鍵結2個糖而成之糖苷，可列舉鳶尾黃素-7-O-木糖葡萄糖苷(tectorigenin-7-O-xylosylglucoside)。此處，所謂於TGE上鍵結n個糖，可鍵結n個糖之連結體，亦可鍵結於鳶尾苷之不同部位之糖之合計數為n個。所謂鳶尾苷，係於TGE之7位上鍵結1個葡萄糖而成之糖苷，例如亦表示為鳶尾黃素7-O-β-D-葡萄糖苷或7-(β-D-吡喃葡萄糖氧基)-4',5-二羥基-6-甲氧基異黃酮(7-(β-D-Glucopyranosyloxy)-4',5-dihydroxy-6-methoxyisoflavone)等。所謂鳶尾苷4'-葡糖苷，係於TGE之4位上鍵結1個葡萄糖而成之糖苷，例如亦稱為鳶尾黃素4'-O-β-D-葡萄糖苷或3-[4-(β-D-吡喃葡萄糖氧基)苯基]-5,7-二羥基-6-甲氧基-4H-1-苯并吡喃-4-酮(3-[4-(β-D-Glucopyranosyloxy)phenyl]-5,7-dihydroxy-6-methoxy-4H-1-benzopyran-4-one)等。鳶尾黃素-7-O-木糖葡萄糖苷係於TGE之7位上鍵結1個葡萄糖，且於該葡萄糖上進而鍵結木糖而成之糖苷，例如亦稱為6''-O-木糖鳶尾苷(6''-O-Xylosyltectoridin)。例如，鳶尾苷(以下亦稱為TD)具有以下化學式之結構。 [化2]又，例如鳶尾黃素-7-O-木糖葡萄糖苷(以下亦稱為TGXG)具有以下化學式之結構。式中，Glu係葡萄糖基，Xyl係木糖基。 [化3]於本發明中，就上述效果優異之方面而言，較佳為TGE糖苷包含於TGE上鍵結1個糖而成之糖苷及於TGE上鍵結2個糖而成之配合體。於TGE糖苷包含於TGE上鍵結1個糖而成之糖苷及於TGE上鍵結2個糖而成之配合體之情形時，於TGE上鍵結1個糖而成之糖苷與於TGE上鍵結2個糖而成之糖苷之質量比較佳為前者：後者為1：0.01以上且7.5以下。該質量比為1：0.01以上有如下優點：能夠獲得本發明之較高作用，並且將TD等之於TGE上鍵結1個糖而成之糖苷之量限制為一定量。又，藉由使該質量比為1：7.5以下，能夠進一步提高上述效果。就該等觀點而言，於TGE上鍵結1個糖而成之糖苷與於TGE上鍵結2個糖而成之糖苷之量比，較佳為前者：後者為1：0.01以上且5以下，尤佳為1：0.1以上且2.5以下。 又，作為TGE糖苷，就更確實地提高上述效果之觀點、及糖苷之獲取容易性等觀點而言，亦較佳為包含TD作為於TGE上鍵結1個糖而成之糖苷，並且包含TGXG作為於TGE上鍵結2個糖而成之配合體，且TD與TGXG之調配比為上述較佳比率1：0.01以上且7.5以下。TD與TGXG之調配比尤佳為1：0.01以上且5以下，尤佳為1：0.1以上且2.5以下。 於本發明中，關於TGE與TGE糖苷之量比，並無特別限制，於將TGE與TGE糖苷以特定比率組合之情形時，能夠獲得尤其優異之效果。於TGE糖苷相對於TGE之質量比為1：10以上之情形時，促進脂肪分解之作用或改善肝功能之作用飛躍性地提高。進而，就使促進脂肪分解之作用或改善肝功能之效果變得尤其優異之觀點而言，TGE與TGR糖苷之質量比更佳為1：13以上，進而較佳為1：16以上，尤佳為1：18以上。又，發現就使減少脂肪、抗肥胖、促進組織分化、提高角質層水分量、提高皮膚彈力或抑制皮膚水分蒸散之效果變得尤其優異之觀點而言，亦較佳為將TGE與TGE糖苷之質量比設為上述特定比率。本發明之組合物可將所合成之鳶尾黃素及鳶尾黃素糖苷組合而調配，亦可為鳶尾科之射干(Belamcanda chinensis)或豆科之葛(粉葛(Pueraria thomsonii)、野葛(Pueraria lobata)、葛藤(Pueraria thunbergiana)等)之花部等植物之萃取物。作為植物之萃取物，尤佳為豆科之葛之花部之萃取物。又，可於植物之萃取物中添加有機合成品而製備，亦可將源自複數種植物之萃取物組合而製備，亦可將有機合成品組合而製備。 關於TGE與TGE糖苷之質量比，就防止由TGE糖苷引起之皮膚炎、嘔吐、下痢、腸胃炎等副作用之觀點而言，較佳為將TGE糖苷之量設為一定以下。就該觀點而言，較佳為將TGE與TGE糖苷之質量比設為較佳為1：500以下、更佳為1：250以下、進而較佳為1：100以下、尤佳為1：50以下。 又，就提高上述效果之觀點而言，作為TGE與TGE糖苷之質量比、以及於TGE上鍵結1個糖而成之糖苷與於TGE上鍵結2個糖而成之糖苷之質量比之組合，較佳為TGE與TGE糖苷之質量比為1：10以上，且於TGE上鍵結1個糖而成之糖苷與於TGE上鍵結2個糖而成之糖苷之質量比為1：0.01以上且7.5以下。該組合中質量比或TGE糖苷之較佳例已如上所述。 本發明之組合物中之TGE及TGE糖苷之量之測定能夠利用HPLC(High performance liquid chromatography，高效液相層析)法定量地或定性地確認。用以供於HPLC之試樣係藉由適當去除雜質或調整濃度等公知之方法製備。 本發明之組合物只要以特定比率包含TGE及TGE糖苷，則會發揮尤其優異之效果，就更進一步提高減少脂肪、抗肥胖、促進組織分化、促進脂肪分解、提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散或改善肝功能之效果之觀點而言，尤佳為包含賦形劑。作為賦形劑，例如可列舉：還原麥芽糖、還原帕拉金糖(reduced palatinose)、糊精、難消化性糊精、纖維素、聚葡萄糖、澱粉、環糊精、乳糖、葡萄糖、砂糖、還原麥芽糖、麥芽糖、山梨糖醇、赤蘚糖醇、木糖醇、乳糖醇、甘露糖醇。其中，就提高本發明之效果之觀點而言，尤佳為還原麥芽糖、還原帕拉金糖、糊精、難消化性糊精、纖維素、聚葡萄糖。 又，就更進一步提高減少脂肪、抗肥胖、促進組織分化、促進脂肪分解、提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散或改善肝功能之效果之觀點而言，本發明之組合物尤佳為包含油脂類。作為油脂類，例如可列舉：紅花油、橄欖油、菜籽油、棕櫚油、芝麻油、大豆油、紫蘇油、亞麻仁油、洋甘菊油、米糠油、小麥胚芽油、牛油樹油脂、蓖麻油、葵花籽油、澳洲胡桃油、糙米胚芽油、山茶油、玉米油、魚油、椰子油、長鏈/中長鏈/短鏈脂肪酸甘油三酯、二醯基甘油酯、以及該等油脂類之氫化物等。其中，就提高本發明之效果之觀點而言，尤佳為紅花油、橄欖油、菜籽油、棕櫚油、芝麻油、大豆油。 本發明之組合物除上述以外，亦可於無損本發明之效果之範圍內含有通常所使用之其他成分。作為此種成分，可列舉：各種結合劑、光澤劑、潤滑劑、穩定劑、稀釋劑、增量劑、增黏劑、乳化劑、抗氧化劑、pH值調整劑、著色料、香料、添加劑等。其他成分之含量能夠視本發明之組合物之形態等而適當地選擇。 本發明之組合物能夠用作外用或經口用。作為外用劑，只要塗抹於皮膚、頭皮等上使用，則並無特別限制，作為其形態，可列舉：軟膏劑、乳霜劑、凝膠劑、洗劑、乳液劑、敷劑、濕敷劑等皮膚外用劑、或注射劑等形態。 又，於將本發明之組合物用作經口劑之情形時，作為其形態，例如可列舉：錠劑、膠囊劑、粉末劑(再者，本發明之粉末包括顆粒、細粒)、液劑、棒狀劑、板狀劑、塊狀劑、固體狀劑、丸狀劑、膏狀劑、乳霜狀劑、膠囊錠狀劑、凝膠狀劑、咀嚼錠狀劑、條狀劑等。該等之中，就更進一步提昇提高角質層水分量、提高皮膚彈力或抑制皮膚水分蒸散之效果之觀點而言，尤佳為錠劑、膠囊劑、粉末劑或液劑之形態。以錠劑、膠囊劑、粉末劑或液劑之形態攝取本發明之組合物之情形與以膏狀或乳霜狀、凝膠狀等其他形態攝取之情形相比，能夠於提高角質層水分量、提高皮膚彈力或抑制皮膚水分蒸散方面獲得更高之效果。其原因並未明確，但認為主要原因在於：根據組合物之形態不同，對於攝取者產生之心理上之影響不同；或者根據形狀不同，TGE及TGE糖苷之體內吸收率不同。 作為將本發明之組合物用作經口劑之情形之具體例，可例示：補充品(supplement)；食品添加劑；填充至寶特瓶、罐、瓶等中之容器裝飲料；用以溶解於水(熱水)、牛奶、果汁等中飲用之即溶粉末飲料等。該等就進食時等容易簡單地飲食，又能夠提高可口性之方面而言較佳。 一般，於本發明之組合物為錠劑、膠囊劑之情形時，就更為提高由本發明帶來之上述各效果之觀點而言，較佳為於其固形物成分中，有效成分包含固形物成分總體之2.0～0.001質量%之TGE，更佳為包含1.0〜0.01質量%，進而較佳為包含0.5〜0.025質量%。又，TGE及TGE糖苷之合計量若於本發明之組合物之固形物成分中例如為0.5質量%以上、尤其是1質量%以上，則上述各效果進一步優異，故而較佳。關於作為錠劑或膠囊劑之本發明之組合物，例如可列舉醫藥品或補充品等。 於本發明之組合物為液劑之情形時，就更為提高由本發明帶來之上述各效果之觀點而言，較佳為於該液劑中，有效成分包含總體之0.00001〜0.5質量%之TGE，更佳為包含0.00005〜0.1質量%，進而較佳為包含0.0001〜0.05質量%。又，TGE及TGE糖苷之合計量若於本發明之組合物之固形物成分中例如為0.0001質量%以上、尤其是0.1質量%以上，則上述各效果進一步優異，故而較佳。關於作為液劑之本發明之組合物，可列舉容器裝飲料等。 又，於本發明之組合物為粉末劑或顆粒劑之情形時，就更為提高由本發明帶來之上述各效果之觀點而言，較佳為於其固形物成分中，有效成分包含固形物成分總體之0.0001〜2.5質量%之TGE，更佳為包含0.001〜2.0質量%，進而較佳為包含0.005～1.5質量%。又，TGE及TGE糖苷之合計量若於本發明之組合物之固形物成分中例如為0.001質量%以上、尤其是0.01質量%以上，則上述各效果進一步優異，故而較佳。關於作為粉末劑之本發明之組合物，可列舉即溶粉末飲料等。 於經口攝取本發明之組合物之情形時，關於其經口投予量，以上述TGE及TGE糖苷之乾燥物合計量計，較佳為成人每天大約5 mg以上且200 mg以下，更佳為10 mg以上且100 mg以下。又，關於本發明之組合物之1次之攝取量，較佳為以上述TGE及TGE糖苷之乾燥物合計量計，成人每天大約15 mg以上且50 mg以下。本發明之組合物能夠以1天量收容於1個容器中，或者分開收容於例如2〜3個之複數個容器中，以使1天之攝取量成為上述攝取量。 作為本發明之組合物之利用形態，具體而言，可列舉：醫藥品(包括準藥品)、化妝品、普通食品、營養功能食品、效能之標示得到特定機構認可之特定保健用食品、功能性標示食品等所謂健康食品等。 本發明之組合物之應用形態中，作為食品，例如可列舉：碳酸飲料、營養飲料、果實飲料、乳酸飲料、果昔(Smoothie)、青汁等飲料；冰淇淋、雪酪、刨冰等冷凍甜點：蕎麥麵、烏冬麵、粉絲、中華麵、即食麵等麵類；飴糖、糖果、口香糖、巧克力、模壓點心、休閒點心、餅乾、果凍、果醬、奶油、烘烤糕點、麵包等點心類；魚糕、火腿、香腸等水產、畜產加工食品；加工乳、醱酵乳、酸奶等乳製品；沙拉油、天婦羅油、人造黃油、蛋黃醬、起酥油、稠奶油、調味醬等油脂及其加工食品；醬料、醬油等調味料；咖喱、燉菜、雞肉雞蛋蓋飯、粥、雜燴、中華蓋飯、豬排蓋飯、炸蝦蓋飯、牛肉蓋飯、肉丁蓋飯、蛋包飯、關東煮、麻婆豆腐、餃子、燒賣、煎肉餅、肉丸子、各種醬料、各種湯等殺菌袋裝食品等。 本發明之組合物不僅能夠經濟且安全地經口攝取，而且根據下述實施例之記載可明確，本發明之組合物藉由含有TGE及TGE糖苷，能夠藉由攝取其而促進從橫紋肌等成肌細胞進行肌肉分化，此外，亦可促進從前成骨細胞向成骨細胞分化，從而能夠促進骨分化。又，本發明之組合物藉由含有TGE及TGE糖苷，能夠藉由攝取其而獲得優異之脂肪分解作用。例如，根據下述實施例之記載可明確，能夠藉由攝取TGE及TGE糖苷而促進分解體脂肪之酶之基因之表現，促進體脂肪分解。 進而，於本發明中，TGE及TGE糖苷有效地提高改善肝功能之作用。具體而言，根據下述實施例之記載可明確，TGE及TGE糖苷能夠提高於無血清培養基中培養之肝細胞之活性(酶活性)，該肝細胞存在各種應力成為誘因而導致容易產生細胞凋亡等問題。即，TGE及TGE糖苷抑制肝細胞之活性降低，即保護肝細胞。如上所述，TGE及TGE糖苷能夠保護肝細胞，因此防止肝細胞之損害，維持或增進肝細胞之活性而改善肝功能。尤其是期待藉由保護肝細胞免受各種應力而防止肝細胞之損傷，從而抑制肝細胞中之天冬胺酸轉胺酶(AST)、丙胺酸轉胺酶(Alanine transaminase，ALT)、γ-谷胺醯轉肽酶(γ-glutamyltransferase，γ-GTP)向血中轉移。 又，藉由攝取TGE及TGE糖苷而能夠獲得提高皮膚之角質層水分量之作用、提高皮膚彈力之作用、抑制皮膚水分蒸散之作用。該等改善膚質之作用係藉由持續地攝取本發明之組合物而顯著地表現，例如於經口攝取開始後4週以內表現，進而藉由8週之持續攝取、尤其是12週之持續攝取而顯著地表現。此處所謂持續攝取，係指持續例如於1～3天內至少攝取1次之狀態。 此種本發明之組合物能夠用作促進肌肉分化用組合物、促進肌肉形成用組合物、促進骨分化用組合物、促進骨形成用組合物、促進脂肪分解用組合物、促進脂質代謝用組合物、減少脂肪用組合物、抗肥胖用組合物、減重用組合物、美容組合物、增加皮膚水分量用組合物、抑制水分蒸散用組合物、提高肌膚彈力用組合物、改善膚質組合物等，此外，亦能夠用作基於預防或抑制肝細胞之脫離酶流出之作用、預防或抑制肝細胞損傷之作用、保護肝細胞之作用、保護或改善肝功能之作用之肝功能改善劑或肝功能改善組合物，能夠用於病毒性肝炎、藥物性肝損傷、自身免疫性肝炎、原發性膽汁性肝硬化、布加氏(Budd-Chiari)症候群等肝功能損傷之治療或預防。如上所述，本發明之組合物能夠用於防止宿醉等與飲酒相關之疾病以外之損傷。 又，本發明之組合物只要於用於促進肌肉分化、促進肌肉形成、促進骨分化、促進骨形成、促進脂肪分解、抗肥胖、減重、美容、改善膚質、改善肝功能之用途之方面，能夠作為製品與其他製品進行區分即可，例如可列舉於本發明之改善肝功能劑之製品之本體、包裝、說明書、宣傳物之任一者上標示出有肝功能改善功能之內容。例如所謂有肝功能改善功能之內容之標示係指：告知如在意肝功能之人、在意肝臟之健康之人等般關心肝臟運作之對象者之標示；或者標示有助於維持增進肝臟之健康，如維持肝臟之健康、輔助肝臟運作、維持健康之肝臟、維持健康之肝臟之功能、降低肝功能酶在正常範圍內偏高之數值等。又，所謂有促進脂肪分解之功能之內容之標示，係指：告知如在意肥胖之人、在意腹部周圍之人、在意體重之人、在意腹部之脂肪(內臟脂肪與皮下脂肪等)之人等般關心體脂肪之對象者之標示；或者標示有助於減少身體之脂肪，如幫助減少體重、幫助減少腹部之脂肪(內臟脂肪與皮下脂肪等)、幫助減少腰圍直徑、輔助消除肥胖、輔助減重等。 本發明之組合物安全，即便持續長期(例如3個月以上)投予(例如於1天中以上述投予量投予之情形)亦無妨。本發明之組合物較佳為如上所述持續使用。又，作為本發明之組合物之投予對象，適宜列舉有AST、ALT、γ-GTP等脫離酶之血中濃度較高之傾向之對象者，從而發揮其保護肝細胞之作用。藉由使此種對象者攝取本發明之組合物，能夠進一步提高防止肝功能降低之效果，且對於防止宿醉等亦有效。 [實施例] 以下，列舉實施例對本發明進一步詳細地進行說明。但，本發明之範圍並不限定於該等實施例。以下，於未特別事先說明之情形時，「%」表示質量%，「份」表示質量份。 [實施例1]作為實施例1之受試物質，使用質量比為鳶尾黃素(TGE)：鳶尾黃素糖苷＝1：21(糖苷之詳細成分以質量比計為TD：TGXG＝1：1.9)之粉末狀組合物。將其分別供於下述(1)〜(3)之順序之肌肉分化試驗、及下述(I)〜(V)之順序之骨分化試驗。 (肌肉分化試驗)(1)將源自小鼠橫紋肌之成肌細胞C2C12(從理化學研究所獲取)利用含10%FBS(Fetal bovine serum，胎牛血清)之DMEM(Dulbecco's minimum essential medium，杜爾伯科最低必需培養基)於5體積%CO2
培養箱中以37℃、濕潤條件進行培養直至成為特定數。其次，去除培養基，利用DPBS(Dulbecco's phosphate buffered saline，杜爾伯科磷酸緩衝液)清洗3次後，利用胰蛋白酶-乙二胺四乙酸(Trypsin-EDTA)將細胞剝離。添加新鮮之含10%FBS之DMEM，使胰蛋白酶反應停止後，將細胞收集至試管，利用離心機以800 rpm進行3分鐘離心而使細胞沈澱。以成為2×104
cells/mL之方式使細胞懸浮於新鮮之含10%FBS之DMEM中，以100 μL/well接種於96孔板中，於5體積%CO2
培養箱中以37℃、濕潤條件預培養2天。 (2)使實施例1之受試物質以TGE及TGE糖苷之總量之濃度成為40 mM之方式溶解於DMSO(Dimethyl sulfoxide，二甲基亞碸)中，將其利用含2%馬血清(HS)之DMEM稀釋至200倍，進而將其以成為特定濃度之2倍之方式利用包含0.5%DMSO之含2%HS之DMEM進行稀釋。此處所謂特定濃度，係指以TGE及TGE糖苷之總量之濃度計為25 μM。 (3)將於(1)中進行過預培養之細胞之培養基去除，添加作為分化誘導培養基之含2%HS之DMEM 50 μL、及於上述(2)中製備之含2%HS之DMEM 50 μL，進而於5%體積CO2
培養箱中以37℃、濕潤條件繼續培養24小時。另一方面，作為對照組，添加作為分化誘導培養基之含2%HS之DMEM 100 μL，以相同方式進行培養。24小時後，去除培養基，使用Rneasy mini(Qiagen公司製造)從細胞純化RNA(ribonucleic acid，核糖核酸)。利用ReverTra Ace(註冊商標)qPCR RT Master Mix with gDNA Remover(東洋紡公司製造)，由經純化之RNA合成cDNA(complementary deoxyribonucleic acid，互補脫氧核糖核酸)。使用GAPDH(glyceraldehyde-3-phosphate dehydrogenase，3-磷酸甘油醛脫氫酶)(Mm_Gapdh_3_SG QuantiTect Primer Assay，Qiagen公司製造)作為內部標準，且使用肌細胞生成素(Myogenin)(Mm_Myog_1_SG QuantiTect primer assay，Qiagen公司製造)之引子、QuantiNOVA SVBR GREEN(Qiagen公司製造)作為測定基因，利用Rotor-Gene Q(Qiagen公司製造)進行PCR(Polymerase chain reaction，聚合酶鏈反應)。PCR之結果係使用Rotor Gene Q Pure Detection(Qiagen公司製造)進行分析所得。關於藉由分析所得之基因表現量，算出將對照組(比較例1)設為100%之相對值。將結果示於圖1中。 一般於成肌細胞中，肌細胞生成素基因之表現被廣泛地用作向肌肉組織之分化誘導標記物。如圖1所示，藉由將具有特定之TGE：TGE糖苷量比之實施例1之組合物添加於分化誘導培養基中之成肌細胞中，肌細胞生成素基因表現量提高。該情況表示於添加有實施例1之組合物之成肌細胞中，向肌肉組織之分化得到促進。 (骨分化試驗)(I)：將源自小鼠頭頂之前成骨細胞MC3T3-E1細胞(從DS Pharma Biomedical公司獲取)利用含10%FBS之MEM(minimum essential medium，最低必需培養基)α培養基(不含抗壞血酸)於5體積%CO2
培養箱中以37℃、濕潤條件進行培養直至成為特定數。去除培養基，利用DPBS清洗2次後，利用胰蛋白酶-乙二胺四乙酸將細胞剝離。添加新鮮之含10%FBS之MEMα培養基，使胰蛋白酶反應停止後，將細胞收集至試管，利用離心機以800 rpm進行3分鐘離心而使細胞沈澱。以成為4×105
cells/mL之方式使細胞懸浮於新鮮之含10%FBS之MEMα培養基中，以100 μL/well接種於96孔板中，於5體積%CO2
、37℃、濕潤條件下預培養1天。 (II)：使抗壞血酸鈉與β甘油磷酸二鈉以分別成為50 μg/mL、10 mM之方式溶解於含10%FBS之MEMα培養基中，製成誘導向骨組織分化之分化誘導培養基。使實施例1之受試物質以按TGE及TGE糖苷之總量之濃度計成為20 mM之方式溶解於DMSO中，將其利用分化誘導培養基稀釋至200倍，進而將其以成為特定濃度之2倍之方式利用包含0.5%DMSO之含10%FBS之MEMα培養基進行稀釋，製成含受試物質之分化誘導培養基。此處所謂特定濃度，係指以TGE及TGE糖苷之總量之濃度計為100 μM。 (III)：將於(I)中進行過預培養之細胞之培養基去除，以100 μl/well添加(II)中製備之含受試物質之分化誘導培養基，2天或3天更換一次培養基，並於5體積%CO2
、37℃、濕潤條件下繼續培養14天。另一方面，作為對照組，將不含受試物質之分化誘導培養基以相同量添加於其他孔中，除此以外，以相同方式進行培養。 (IV)上述(III)之培養後，去除培養基，利用無血清DMEM清洗1次後，添加利用無血清DMEM培養基稀釋至30倍之細胞計數試劑盒-8(Cell Counting-Kit 8)(同仁化學公司製造)150 μL，於37℃下進行保溫直至適當顯色。利用讀板儀(ThermoScientific公司製造)測定450 nm之吸光度(A1)。(V)從(IV)之測定後之培養基去除細胞計數試劑盒-8，依據TRAP/ALP(tartrate-resistant acid phosphatase/alkaline phosphatase，抗酒石酸性磷酸酶/鹼性磷酸酶)染色試劑盒(和光純藥公司製造)之說明書，進行鹼性磷酸酶(ALP)活性測定用之染色。其次，使甘胺酸、無水氯化鎂、氯化鋅以分別成為0.1 M、1 mM、1 mM之方式溶解於超純水中，利用氫氧化鈉將pH值調整至10.4，製成甘胺酸緩衝液。使一粒4-硝基苯基磷酸二鈉鹽六水合物5 mg錠劑(Sigma Aldrich公司製造)溶解於所獲得之甘胺酸緩衝液5 ml中。將所得之液體添加至ALP染色後之各孔中150 μL。以37℃加溫30分鐘後，將上清液100 μL移至分析板(assay plate)(AGC Techno Glass公司製造)中，利用讀板儀對405 nm之吸光度(A2)進行測定。使用上述中所測得之吸光度A1及A2，藉由下述式求出ALP活性。關於由ALP活性＝A2/A1 所獲得之ALP活性之值，將對照組(比較例1)設為100%而算出相對值。將結果示於圖2中。 一般，ALP活性被廣泛地用作MC3T3-E1細胞向成骨細胞之分化誘導標記物。如圖2所示，藉由將具有特定之TGE：TGE糖苷量比之實施例1之組合物添加於分化誘導培養基中之MC3T3-E1細胞中，ALP活性提高。該情況表示於添加有實施例1之組合物之MC3T3-E1細胞中，成骨分化得到促進。 [實施例2-1〜2-8、比較例2]作為受試物質，使用將TD、TGXG及TGE以下述表1之記載進行調配所得者。表1中之TGE糖苷係指TD及TGXG之合計量。將該等實施例及比較例之受試物質供於下述(i)～(viii)之順序之(脂肪分解酶表現試驗)。
(表中，數字表示質量比) (脂肪分解酶表現試驗)利用下述方法測定激素敏感性脂肪酶(HSL)之基因表現。(i)於37℃、5體積%CO2
培養箱內，使用75 cm2
燒瓶，將供分化為脂肪細胞之小鼠成纖維(fibroblast)細胞(3T3-L1)於預培養培養基中進行培養。作為預培養培養基，使用10%FBS-DMEM。(ii)使藉由胰蛋白酶處理而浮游之細胞懸浮於10%FBS-DMEM中，以2×103
cells/well之細胞密度接種至96孔板之各孔中。將接種後之細胞於37℃、5體積%CO2
培養箱內預培養2天。(iii)去除培養基，以100 μL/well添加預先以分別包含1 μM之地塞米松、1 mM之3-異丁基-1-甲基黃嘌呤(3-Isobutyl-1-methylxanthine)、20 μg/mL之胰島素之方式製備之10%FBS-DMEM(向脂肪細胞之分化誘導培養基)，於37℃、5體積%CO2
培養箱內培養2天。(iv)從各孔去除一半培養基，更換為預先以包含20 μg/mL之胰島素之方式製備之10%FBS-DMEM(脂肪細胞分化維持培養基)，進而於37℃、5體積%CO2
培養箱內培養5天。在此期間，每隔2天將培養基之一半更換為包含胰島素之10%FBS-DMEM。(v)從各孔將培養基完全去除，以100 μL/well(TGE及TGE糖苷之總量之濃度為100 μM)添加預先以受試物質成為100 μg/mL之方式製備之10%FBS-DMEM，進而於37℃、5體積%CO2
培養箱內繼續培養48小時。(vi)將培養基完全去除，利用經冰浴冷卻之DPBS清洗2次後，以100 μL/well添加ISOGEN(NIPPON GENE公司製造)，按照使用說明書將總RNA進行純化。(vii)利用ReverTra AceR qPCR RT Master Mix with gDNA Remover，依據使用說明書，從(vi)中所獲得之總RNA合成cDNA。(viii)使用QuantiNova SYBR Green PCR試劑盒，將(vii)中所獲得之cDNA作為模板，進行RT-PCR(Reverse Transcription-Polymerase Chain Reaction，逆轉錄-聚合酶鏈反應)。作為引子，使用作為HSL基因表現用之Mm_Lipe_1_SG QuantiTect Primer Assay。內部標準使用Mm_Actb_1_SG QuantiTect Primer Assay進行Actb。關於HSL基因表現量，將以比較例2(對照組)之基因表現量為1時之實施例2-1〜2-8之相對值示於圖3中。 如圖3所示，藉由使用各實施例之受試物質，激素敏感性脂肪酶(HSL)之基因表現量大幅地高於比較例2。已知激素敏感性脂肪酶(HSL)為中性脂肪分解酶，且存在於脂肪細胞內，將甘油三酯分解為游離脂肪酸與甘油，並使游離脂肪酸釋出於血液中。因此，判明本發明之組合物藉由促進激素敏感性脂肪酶(HSL)基因表現，而有效地促進脂肪細胞中之脂肪分解。 (肝功能改善試驗)[實施例3-1～3-16以及比較例3](1)細胞培養 於37℃、5體積%CO2
培養箱內，使用75 cm2
燒瓶，藉由含10%胎牛血清(FBS)之培養基培養源自人類肝癌之細胞(HepG2，ATCC公司製造)。其次，將藉由胰蛋白酶處理而浮游之細胞以4.0×104
cells/well之細胞密度從75 cm2
燒瓶接種至96孔板之各孔中。其後，於37℃、5體積%CO2
培養箱內預培養24小時。從各孔去除培養基後，以100 μL/well添加製備成特定濃度之含受試物質之培養基，於CO2
培養箱內培養24小時。作為含受試物質之培養基中之培養基，使用無血清DMEM。作為受試物質，使用將TD、TGXG及TGE以下述表2之記載調配所得者。表2中之TGE糖苷係指TD及TGXG之合計量。作為上述特定濃度，以TD、TGXG及TGE之總量之濃度計設為400 μg/ml。比較例3為對照組(control)，使用含0.5%DMSO之無血清DMEM代替含受試物質之培養基。
＜細胞活性測定＞上述24小時培養後，去除培養基，將利用無血清DMEM稀釋至30體積倍之細胞計數試劑盒-8溶液(同仁化學製造)以每孔150 μl之方式添加至各孔中。於37℃、5體積%CO2
培養箱內進行靜置而適度地顯色後，測定450 nm下之吸光度。基於所獲得之資料，算出相對於對照組之百分率(% of control)。將由相對於對照組之百分率(% of control)＝(樣本資料(Data sample)－空白資料(Data blank))/(對照資料(Data control)－空白資料(Data blank))×100所獲得之結果示於圖4。於圖4中記載平均值及標準偏差。 根據圖4所示之結果可知，TGE及其糖苷之量比特定之鳶尾黃素類具有保護於無血清DMEM中培養之肝細胞之活性之優異作用。因此，藉由使用該質量比特定之鳶尾黃素類，能夠保護肝細胞而改善肝功能。 [實施例4]作為實施例4之受試物質，使用質量比為TGE：TGE糖苷＝1：22.19(TD：TGXG＝1：1.81)之粉末狀組合物43.14 mg(TGE及TGE糖苷之總量)。將其供於下述美容試驗。 (美容試驗)將健康正常之2名女性(平均年齡19.7歲)作為受試者。請受試者每天早晨攝取溶解於水中之實施例4之受試物質。將其持續12週。於攝取開始前、攝取開始後(4、8、12週後)，分別利用下述測定裝置測定左頰部(用直線連接外眼角與鼻翼時之中央附近)之角質層水分量、水分蒸散量、彈力。將角質層水分量之平均值示於圖5，將水分蒸散量之平均值示於圖6，將彈力之平均值示於圖7。 •角質層水分量：利用皮表角質層水分量測定裝置SKICON-200EX(I.B.S.公司製造)進行測定。該皮表角質層水分量測定裝置係將皮膚之導電率(conductance，單位：μS)作為角質層之水分量進行評價者。 •水分蒸散量：利用Tewameter TH300(Courage＋Khazaka公司製造)進行測定。 •彈力：利用皮膚黏彈力測定裝置Cutometer MPA580(Courage＋
Khazaka公司製造)進行測定。關於彈力，使用恢復彈性比率作為能夠利用作為測定器之Cutometer獲得之皮膚黏彈性值之測定參數，該恢復彈性比率係以負壓吸引測定部之皮膚一定時間，對被吸入之高度及從吸引部頂點恢復之皮膚之位移量進行分析而求出。恢復彈性比率越接近1.00，可謂皮膚之彈力性越高。該等測定係利用機器所附帶之說明書所記載之標準方法實施。 根據圖5〜圖7之結果，判明隨著具有特定之TGE：TGE糖苷量比之本發明之組合物攝取時間變長，肌膚之角質層水分量提高，水分蒸散量得到抑制，且彈力提高。 以下列舉本發明之組合物之調配例，但下述調配例並不限定本發明。 調配例1〜10：錠劑 以成為表3之比例之方式調配各成分，依據慣例進行打錠而製造調配例1〜10之錠劑(1錠250 mg)。調配例1〜10之錠劑於尤其良好地發揮提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散等本發明之效果之方面而言優異。 [表3]
調配例11〜20：軟膠囊 藉由以成為表4之比例之方式製備軟膠囊之內容液並填充於膠囊皮膜中，而製造軟膠囊(1粒300 mg)。將膠囊皮膜液流延而製成膜，並且將內容液填充於內部進行熱密封，並使成形之軟膠囊進行乾燥而製成膠囊。調配例11〜20之軟膠囊於尤其良好地發揮提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散等本發明之效果之方面而言優異。
調配例21〜30：粉末(顆粒劑) 依據表5，將各成分投入至Flow Coater NFLO-200型流動層造粒機(Freund Corporation(股)製造)中，於氣流下混合數分鐘。藉由對其噴霧水而進行造粒。其次，利用30目之篩子篩選所獲得之造粒物而製造顆粒劑。調配例21〜22之顆粒劑於尤其良好地發揮提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散等本發明之效果之方面而言優異。
調配例21-22：液劑(液體飲料) 以表6之比例調配各成分而製成液體飲料。調配例31〜40之液體飲料於尤其良好地發揮提高角質層水分量、提高皮膚彈力、抑制皮膚水分蒸散等本發明之效果之方面而言優異。 Hereinafter, the present invention will be described in more detail with reference to embodiments of the present invention, but the present invention is not limited to these. Hereinafter, in the present invention, a composition comprising irisin and irisin glycoside, and a composition for reducing fat, a composition for anti-obesity, a composition for promoting tissue differentiation, and a composition for promoting lipolysis comprising these are used The composition, the composition for increasing the moisture content of the stratum corneum, improving the skin elasticity or inhibiting the transpiration of skin water, and the composition for improving the liver function are collectively referred to as "the composition of the present invention". (Irisin) The irisin used in the present invention is represented by the molecular formula C 16 H 12 O 6 , also known as 5,7-dihydroxy-3-(4-hydroxyphenyl)-6-methoxy yl-4H-1-benzopyran-4-one (5,7-Dihydroxy-3-(4-hydroxyphenyl)-6-methoxy-4H-1-benzopyran-4-one) or 6 - methoxy- 5,7-Dihydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one (6 - Methoxy-5,7-dihydroxy-3-(4-hydroxyphenyl)-4H- 1-benzopyran-4-one). Hereinafter, irisin is also described as TGE. The chemical formula of irisin is shown below. [hua 1] (Iris flavin glycosides) Iris flavin glycosides (hereinafter also referred to as TGE glycosides) are those obtained by substituting 1 or 2 hydrogens in the above-mentioned TGE with sugars. Sugar, sorbose, galactose, celose, rhamnose, one or two or more sugars. From the viewpoint of enhancing the effects of the present invention, such as promoting tissue differentiation, promoting lipolysis, improving liver function, and improving skin texture, it is preferable to bond one or more sugars selected from glucose and xylose to TGE. become. Among the glycosides, these sugars are usually bonded to the hydroxyl group at the 4-position and/or the 7-position of TGE, and in terms of obtaining the above-mentioned effects more reliably or the easiness of obtaining the glycoside, it is preferably the 7-position. hydroxyl bonder. The number of sugars in TGE glycosides (also referred to as the number of bonds of sugar) can be, for example, one or more, and from the viewpoint of improving the above effect, the number of bonds of sugar is preferably 1 to 3, more preferably 1~2 pieces. The number of sugars here refers to the number of monosaccharide units. As TGE glycosides, only one type may be used, but two or more types are preferably used in combination. In the composition of the present invention, TGE and TGE glycosides can be organic synthetic products, and can also be extracted from plants and the like. TGE and TGE glycosides may be in a solid state such as powder, or may be in a state of being dissolved in water or an organic solvent. TGE and TGE glycosides may exist only in the form of a mixture of these, or may be in a state of being dissolved, dispersed or mixed into other substances. As a glycoside which couple|bonded one sugar to TGE, irisin (Tectoridin) and irisin 4'-glucoside are mentioned, for example. Moreover, tectorigenin-7-O-xylosylglucoside is mentioned as a glycoside which couple|bonded two sugars to TGE. Here, the so-called n sugars are bonded to TGE, which may be a linker of n sugars, and the total number of sugars which may be bonded to different parts of irisin is n. The so-called irisin is a glycoside formed by bonding a glucose to the 7-position of TGE, for example, it is also expressed as irisin 7-O-β-D-glucoside or 7-(β-D-glucopyranosyloxy base)-4', 5-dihydroxy-6-methoxyisoflavone (7-(β-D-Glucopyranosyloxy)-4', 5-dihydroxy-6-methoxyisoflavone) and so on. The so-called irisin 4'-glucoside is a glycoside formed by bonding a glucose to the 4-position of TGE, such as irisin 4'-O-β-D-glucoside or 3-[4- (β-D-glucopyranosyloxy)phenyl]-5,7-dihydroxy-6-methoxy-4H-1-benzopyran-4-one (3-[4-(β-D -Glucopyranosyloxy)phenyl]-5,7-dihydroxy-6-methoxy-4H-1-benzopyran-4-one) and so on. Irisin-7-O-xylose glucoside is a glycoside formed by bonding a glucose on the 7th position of TGE, and then bonding xylose on the glucose, for example, it is also called 6''-O-wood Glycosin (6''-O-Xylosyltectoridin). For example, irisin (hereinafter also referred to as TD) has the structure of the following chemical formula. [hua 2] Also, for example, irisin-7-O-xyloglucoside (hereinafter also referred to as TGXG) has the structure of the following chemical formula. In the formula, Glu is a glucosyl group, and Xyl is a xylosyl group. [hua 3] In the present invention, it is preferable that the TGE glycoside includes a glycoside in which one sugar is bonded to TGE and a complex in which two sugars are bonded to TGE, from the viewpoint of being excellent in the above effects. When the TGE glycoside includes a glycoside formed by bonding 1 sugar on TGE and a complex formed by bonding 2 sugars on TGE, the glycoside formed by bonding 1 sugar on TGE and the complex formed by bonding 1 sugar on TGE The mass ratio of the glycoside formed by bonding two sugars is preferably the former: the latter is 1:0.01 or more and 7.5 or less. The mass ratio of 1:0.01 or more has the advantages that the higher effect of the present invention can be obtained, and the amount of the glycoside such as TD, which is formed by bonding one sugar to TGE, is limited to a certain amount. Moreover, by making this mass ratio 1:7.5 or less, the said effect can be improved further. From these viewpoints, the ratio of the amount of the glycoside formed by bonding one sugar to TGE and the amount of the glycoside formed by bonding two sugars to TGE is preferably the former: the latter is 1:0.01 or more and 5 or less , particularly preferably 1:0.1 or more and 2.5 or less. In addition, as TGE glycosides, it is also preferable to include TD as a glycoside formed by bonding one sugar to TGE, and to include TGXG from the viewpoints of more reliably enhancing the above-mentioned effects, and from the viewpoints of the availability of glycosides. As a complex in which two sugars are bonded to TGE, the blending ratio of TD and TGXG is the above-mentioned preferred ratio of 1:0.01 or more and 7.5 or less. The mixing ratio of TD and TGXG is more preferably 1:0.01 or more and 5 or less, particularly preferably 1:0.1 or more and 2.5 or less. In the present invention, the amount ratio of TGE and TGE glycoside is not particularly limited, and when TGE and TGE glycoside are combined in a specific ratio, particularly excellent effects can be obtained. When the mass ratio of TGE glycoside to TGE is 1:10 or more, the effect of promoting lipolysis or the effect of improving liver function is dramatically increased. Furthermore, the mass ratio of TGE to TGR glycoside is more preferably 1:13 or more, more preferably 1:16 or more, particularly preferably from the viewpoint of making the effect of promoting lipolysis or the effect of improving liver function particularly excellent. 1:18 or more. In addition, it was found that the combination of TGE and TGE glycosides is also preferable from the viewpoint of making the effects of reducing fat, anti-obesity, promoting tissue differentiation, increasing the moisture content of the stratum corneum, improving skin elasticity, or inhibiting skin moisture transpiration particularly excellent. The mass ratio is set to the above-mentioned specific ratio. The composition of the present invention can be prepared by combining the synthesized irisin and irisin glycosides, and can also be Belamcanda chinensis of Iridaceae or kudzu (Pueraria thomsonii), kudzu (Pueraria) lobata), kudzu (Pueraria thunbergiana, etc.) flower parts and other plant extracts. As a plant extract, it is especially the extract of the kudzu flower of the legume family. Moreover, it can be prepared by adding an organic synthetic product to a plant extract, and it can also be prepared by combining the extracts derived from a plurality of plants, and it can also be prepared by combining an organic synthetic product. Regarding the mass ratio of TGE to TGE glycoside, from the viewpoint of preventing side effects such as dermatitis, vomiting, diarrhea, and gastroenteritis caused by TGE glycoside, it is preferable to set the amount of TGE glycoside to a certain value or less. From this viewpoint, the mass ratio of TGE and TGE glycoside is preferably 1:500 or less, more preferably 1:250 or less, still more preferably 1:100 or less, particularly preferably 1:50 the following. Also, from the viewpoint of improving the above-mentioned effects, the mass ratio of TGE and TGE glycosides, and the mass ratio of the glycosides formed by bonding one sugar to TGE and the glycosides formed by bonding two sugars to TGE Combination, preferably the mass ratio of TGE and TGE glycoside is 1:10 or more, and the mass ratio of the glycoside formed by bonding 1 sugar on TGE to the glycoside formed by bonding 2 sugars on TGE is 1: 0.01 or more and 7.5 or less. Preferred examples of the mass ratio or TGE glycosides in the combination are as described above. The measurement of the amount of TGE and TGE glycosides in the composition of the present invention can be quantitatively or qualitatively confirmed by HPLC (High performance liquid chromatography, high performance liquid chromatography). A sample for HPLC is prepared by a known method such as appropriate removal of impurities or concentration adjustment. As long as the composition of the present invention contains TGE and TGE glycoside in a specific ratio, it exhibits particularly excellent effects, and further improves fat reduction, anti-obesity, promotion of tissue differentiation, promotion of lipolysis, improvement of stratum corneum moisture content, and improvement of skin elasticity. From the viewpoint of the effect of inhibiting skin water evapotranspiration or improving liver function, it is particularly preferable to contain an excipient. Examples of excipients include reduced maltose, reduced palatinose, dextrin, indigestible dextrin, cellulose, polydextrose, starch, cyclodextrin, lactose, glucose, sugar, reduced Maltose, Maltose, Sorbitol, Erythritol, Xylitol, Lactitol, Mannitol. Among them, from the viewpoint of enhancing the effect of the present invention, reduced maltose, reduced palatinose, dextrin, indigestible dextrin, cellulose, and polydextrose are particularly preferred. Also, from the viewpoint of further enhancing the effects of reducing fat, anti-obesity, promoting tissue differentiation, promoting lipolysis, increasing the moisture content of the stratum corneum, improving skin elasticity, inhibiting skin moisture transpiration, or improving liver function, the composition of the present invention It is especially preferable to contain fats and oils. Examples of oils and fats include safflower oil, olive oil, rapeseed oil, palm oil, sesame oil, soybean oil, perilla oil, linseed oil, chamomile oil, rice bran oil, wheat germ oil, shea butter, and castor oil , sunflower oil, macadamia oil, brown rice germ oil, camellia oil, corn oil, fish oil, coconut oil, long-chain/medium-chain/short-chain fatty acid triglycerides, diacylglycerols, and any of these oils hydride, etc. Among them, safflower oil, olive oil, rapeseed oil, palm oil, sesame oil, and soybean oil are particularly preferred from the viewpoint of enhancing the effects of the present invention. In addition to the above, the composition of the present invention may contain other commonly used components within a range that does not impair the effects of the present invention. Examples of such components include various binders, glossing agents, lubricants, stabilizers, diluents, extenders, tackifiers, emulsifiers, antioxidants, pH adjusters, colorants, fragrances, additives, and the like . The content of other components can be appropriately selected depending on the form and the like of the composition of the present invention. The composition of the present invention can be used for external use or oral use. The external preparation is not particularly limited as long as it is used on the skin, scalp, etc., and examples thereof include ointments, creams, gels, lotions, emulsions, compresses, and wet compresses Forms such as external skin preparations or injections. When the composition of the present invention is used as an oral preparation, examples of the form include lozenges, capsules, powders (in addition, the powder of the present invention includes granules and fine particles), liquid tablets, sticks, tablets, blocks, solids, pills, pastes, creams, capsules, gels, chewing tablets, bars, etc. . Among these, from the viewpoint of further enhancing the effect of increasing the moisture content of the stratum corneum, improving skin elasticity, or inhibiting skin moisture transpiration, the form of a tablet, a capsule, a powder or a liquid is particularly preferred. When ingesting the composition of the present invention in the form of lozenge, capsule, powder or liquid, compared with the case of ingesting it in other forms such as paste, cream, gel, etc., the moisture content of the stratum corneum can be increased. , Improve skin elasticity or inhibit skin moisture evaporation to obtain higher effect. The reason for this is not clear, but it is considered that the main reasons are: the psychological effects on the ingested person differ depending on the form of the composition; or the absorption rate of TGE and TGE glycosides in vivo differs depending on the form. Specific examples of the case where the composition of the present invention is used as an oral preparation include: supplements; food additives; beverages filled in containers such as PET bottles, cans, and bottles; (hot water), milk, juice and other instant powder beverages. These are preferable in that it is easy to eat and drink easily, and the palatability can be improved. In general, when the composition of the present invention is a tablet or a capsule, from the viewpoint of further enhancing the above-mentioned effects brought about by the present invention, it is preferable that the active ingredient contains the solid content in the solid content. 2.0-0.001 mass % of TGE of the whole component is more preferable to contain 1.0-0.01 mass %, and it is still more preferable to contain 0.5-0.025 mass %. Moreover, when the total amount of TGE and TGE glycosides is, for example, 0.5 mass % or more, especially 1 mass % or more in the solid content of the composition of the present invention, since the above-mentioned effects are further excellent, it is preferable. The composition of the present invention as a tablet or capsule includes, for example, pharmaceuticals, supplements, and the like. When the composition of the present invention is a liquid preparation, from the viewpoint of further enhancing the above-mentioned effects brought about by the present invention, it is preferable that the active ingredient in the liquid preparation contains 0.00001 to 0.5 mass % of the total. TGE is more preferably contained in 0.00005 to 0.1 mass %, and still more preferably contained in 0.0001 to 0.05 mass %. Moreover, when the total amount of TGE and TGE glycosides is, for example, 0.0001 mass % or more, particularly 0.1 mass % or more, in the solid content of the composition of the present invention, since the above-mentioned effects are further excellent, it is preferable. As for the composition of the present invention as a liquid agent, a container-packed drink etc. are mentioned. In addition, when the composition of the present invention is a powder or granule, from the viewpoint of further enhancing the above-mentioned effects brought about by the present invention, it is preferable that the active ingredient contains the solid content in the solid content. 0.0001-2.5 mass % of TGE of the whole component is more preferable to contain 0.001-2.0 mass %, and it is still more preferable to contain 0.005-1.5 mass %. Moreover, when the total amount of TGE and TGE glycosides is, for example, 0.001 mass % or more, especially 0.01 mass % or more, in the solid content of the composition of the present invention, since the above-mentioned effects are further excellent, it is preferable. As the composition of the present invention as a powder agent, an instant powder drink etc. are mentioned. In the case of oral ingestion of the composition of the present invention, the oral dosage is preferably about 5 mg or more and 200 mg or less per day for adults, more preferably in terms of the total amount of the above-mentioned dry matter of TGE and TGE glycosides 10 mg or more and 100 mg or less. In addition, the one-time ingestion of the composition of the present invention is preferably about 15 mg or more and 50 mg or less per day for an adult, based on the total amount of the above-mentioned TGE and TGE glycosides in dry matter. The composition of the present invention can be contained in one container for one day, or can be contained in a plurality of containers such as 2 to 3 separately, so that the one-day intake becomes the above-mentioned intake. Specific examples of the utilization form of the composition of the present invention include pharmaceuticals (including quasi-drugs), cosmetics, general foods, nutritional functional foods, specific health-care foods whose efficacy labels are approved by specific agencies, and functional labels. Food and other so-called health food, etc. In the application form of the composition of the present invention, as food, for example, beverages such as carbonated beverages, nutritional beverages, fruit beverages, lactic acid beverages, smoothies, and green juices; frozen desserts such as ice cream, sherbet, and shaved ice: Soba noodles, udon noodles, vermicelli noodles, Chinese noodles, instant noodles and other noodles; caramel, candy, chewing gum, chocolate, molded desserts, leisure snacks, biscuits, jelly, jam, cream, baked cakes, bread and other desserts; fish Cakes, hams, sausages and other aquatic and livestock processed foods; processed milk, fermented milk, yogurt and other dairy products; salad oil, tempura oil, margarine, mayonnaise, shortening, heavy cream, seasoning sauce and other oils and their processing Food; sauces, soy sauce and other seasonings; curry, stew, chicken and egg rice bowl, porridge, chowder, Chinese rice bowl, pork chop rice bowl, fried shrimp rice bowl, beef rice bowl, diced pork rice bowl, omelette rice, oden , Mapo tofu, dumplings, shumai, pan-fried meat patties, meatballs, various sauces, various soups and other sterilized bagged foods. The composition of the present invention can not only be ingested economically and safely, but it is also clear from the description of the following examples that the composition of the present invention contains TGE and TGE glycoside, and can promote the formation of striated muscle and the like by ingesting them. Muscle cells undergo muscle differentiation and, in addition, can promote differentiation from pre-osteoblasts to osteoblasts, thereby promoting bone differentiation. Moreover, since the composition of this invention contains TGE and TGE glycoside, it can acquire the outstanding lipolysis effect|action by ingesting them. For example, it is clear from the description of the following examples that the body fat decomposition can be promoted by ingesting TGE and TGE glycosides to promote the expression of genes for enzymes that decompose body fat. Furthermore, in the present invention, TGE and TGE glycosides effectively enhance the effect of improving liver function. Specifically, according to the descriptions in the following examples, it is clear that TGE and TGE glycosides can increase the activity (enzymatic activity) of hepatocytes cultured in serum-free medium, and that various stresses in the hepatocytes can induce apoptosis and cause apoptosis death and other issues. That is, TGE and TGE glycosides inhibit the decrease in the activity of hepatocytes, that is, protect hepatocytes. As mentioned above, TGE and TGE glycosides can protect liver cells, thereby preventing liver cell damage, maintaining or enhancing liver cell activity and improving liver function. In particular, it is expected to prevent liver cell damage by protecting liver cells from various stresses, thereby inhibiting aspartate transaminase (AST), alanine transaminase (ALT), γ- Glutamate transpeptidase (γ-glutamyltransferase, γ-GTP) is transferred to the blood. In addition, by ingesting TGE and TGE glycosides, the effect of increasing the moisture content of the stratum corneum of the skin, the effect of improving skin elasticity, and the effect of inhibiting the transpiration of skin water can be obtained. These skin texture-improving effects are remarkably manifested by continuous ingestion of the composition of the present invention, for example, within 4 weeks after the initiation of oral ingestion, and further by continuous ingestion for 8 weeks, especially 12 weeks. ingested and manifested significantly. The term "continuous ingestion" here refers to a state of continuous ingestion, for example, at least once in 1 to 3 days. The composition of the present invention can be used as a composition for promoting muscle differentiation, a composition for promoting muscle formation, a composition for promoting bone differentiation, a composition for promoting bone formation, a composition for promoting fat decomposition, and a composition for promoting lipid metabolism composition, composition for fat reduction, composition for anti-obesity, composition for weight loss, cosmetic composition, composition for increasing skin moisture content, composition for inhibiting water transpiration, composition for improving skin elasticity, composition for improving skin texture In addition, it can also be used as a liver function improving agent or liver function based on the effect of preventing or inhibiting the efflux of detachment enzymes from hepatocytes, the effect of preventing or inhibiting liver cell damage, the effect of protecting hepatocytes, and the effect of protecting or improving liver function. The function-improving composition can be used for the treatment or prevention of liver function damage such as viral hepatitis, drug-induced liver injury, autoimmune hepatitis, primary biliary cirrhosis, and Budd-Chiari syndrome. As described above, the composition of the present invention can be used to prevent damages other than alcohol-related diseases such as hangovers. In addition, the composition of the present invention only needs to be used for the purposes of promoting muscle differentiation, promoting muscle formation, promoting bone differentiation, promoting bone formation, promoting lipolysis, anti-obesity, weight loss, beauty, improving skin texture, and improving liver function. It can be used as a product to distinguish it from other products, for example, it can be listed in any one of the body, package, instruction manual and publicity material of the product of the liver function improving agent of the present invention, which is marked with the content of liver function improving function. For example, the so-called label with the content of improving liver function refers to a label that informs those who care about liver function, such as those who care about liver function, those who care about liver health, etc.; or label that helps maintain and improve liver health, Such as maintaining the health of the liver, assisting the functioning of the liver, maintaining a healthy liver, maintaining the function of a healthy liver, reducing the values of liver function enzymes that are high in the normal range, etc. In addition, the so-called indication of the content of the function of promoting fat decomposition refers to: those who care about obesity, those who care about the abdomen, those who care about weight, those who care about abdominal fat (visceral fat, subcutaneous fat, etc.) Labels for those who are generally concerned about body fat; or labels that help reduce body fat, such as helping to reduce weight, helping to reduce abdominal fat (visceral fat and subcutaneous fat, etc.) Wait. The composition of the present invention is safe even if it is continuously administered for a long time (for example, more than 3 months) (for example, in the case of administering the above-mentioned dose in one day). The compositions of the present invention are preferably used continuously as described above. Moreover, as the subject to which the composition of the present invention is administered, there is preferably a subject whose blood concentration of dezymes such as AST, ALT, and γ-GTP tends to be high, thereby exerting its protective effect on hepatocytes. By ingesting the composition of the present invention in such a subject, the effect of preventing a decrease in liver function can be further enhanced, and it is also effective for preventing hangovers and the like. [Examples] Hereinafter, the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these Examples. Hereinafter, "%" represents mass %, and "part" represents mass part unless otherwise specified. [Example 1] As the test substance of Example 1, the mass ratio of irisin (TGE): irisin glycosides = 1:21 (the detailed components of the glycosides in terms of mass ratios are TD: TGXG = 1:1.9 ) powder composition. They were respectively used for the muscle differentiation test in the following (1)~(3) order, and the bone differentiation test in the following (1)~(V) order. (Muscle differentiation test) (1) Myoblasts C2C12 (obtained from the Institute of Physics and Chemistry) derived from mouse striated muscle were treated with DMEM (Dulbecco's minimum essential medium, Dulbecco's minimum essential medium) containing 10% FBS (Fetal bovine serum, fetal bovine serum). Albuquerque Minimum Essential Medium) was cultured in a 5 vol% CO2 incubator at 37°C under humidified conditions until a specific number was reached. Next, the medium was removed, and after washing three times with DPBS (Dulbecco's phosphate buffered saline), the cells were detached with trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA). After adding fresh DMEM containing 10% FBS to stop the trypsin reaction, the cells were collected into a test tube, and the cells were pelleted by centrifugation at 800 rpm for 3 minutes in a centrifuge. The cells were suspended in fresh DMEM containing 10% FBS in a manner of 2×10 4 cells/mL, seeded in a 96-well plate at 100 μL/well, and incubated in a 5 vol% CO 2 incubator at 37°C, Humid conditions were pre-incubated for 2 days. (2) The test substance of Example 1 was dissolved in DMSO (Dimethyl sulfoxide, dimethyl sulfoxide) so that the concentration of the total amount of TGE and TGE glycosides was 40 mM, and the test substance containing 2% horse serum ( HS) was diluted 200-fold with DMEM containing 2% HS containing 0.5% DMSO so as to be 2-fold the specific concentration. The so-called specific concentration here means that the concentration of the total amount of TGE and TGE glycosides is 25 μM. (3) The medium of the precultured cells in (1) was removed, and 50 μL of DMEM containing 2% HS as a differentiation induction medium, and 50 DMEM containing 2% HS prepared in (2) above were added. μL, and further cultured in a 5% volume CO 2 incubator at 37°C under humid conditions for 24 hours. On the other hand, as a control group, 100 μL of DMEM containing 2% HS was added as a differentiation induction medium, and culture was performed in the same manner. After 24 hours, the medium was removed, and RNA (ribonucleic acid, ribonucleic acid) was purified from the cells using RNeasy mini (manufactured by Qiagen). Using ReverTra Ace (registered trademark) qPCR RT Master Mix with gDNA Remover (manufactured by Toyobo Co., Ltd.), cDNA (complementary deoxyribonucleic acid, complementary deoxyribonucleic acid) was synthesized from the purified RNA. GAPDH (glyceraldehyde-3-phosphate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase) (Mm_Gapdh_3_SG QuantiTect Primer Assay, manufactured by Qiagen) was used as an internal standard, and Myogenin (Mm_Myog_1_SG QuantiTect primer assay, Qiagen) was used PCR (Polymerase chain reaction, polymerase chain reaction) was performed using Rotor-Gene Q (manufactured by Qiagen) as a primer of QuantiNOVA SVBR GREEN (manufactured by Qiagen) as the measurement gene. The results of PCR were analyzed using Rotor Gene Q Pure Detection (manufactured by Qiagen). The relative value of the control group (Comparative Example 1) as 100% was calculated with respect to the gene expression level obtained by the analysis. The results are shown in FIG. 1 . Generally in myoblasts, the expression of the myogenin gene is widely used as a marker for induction of differentiation into muscle tissue. As shown in FIG. 1 , by adding the composition of Example 1 with a specific TGE:TGE glycoside ratio to myoblasts in the differentiation induction medium, the expression level of the myogenin gene was increased. This indicates that the myoblasts to which the composition of Example 1 was added were promoted to differentiate into muscle tissue. (Bone differentiation test) (I): The MC3T3-E1 cells (obtained from DS Pharma Biomedical, Inc.) derived from mouse head pre-osteoblasts were treated with 10% FBS-containing MEM (minimum essential medium, minimum essential medium) α medium ( Without ascorbic acid) in a 5 vol% CO2 incubator at 37°C, humidified conditions until the specified number. The medium was removed, and after washing twice with DPBS, the cells were detached with trypsin-ethylenediaminetetraacetic acid. After adding fresh MEMα medium containing 10% FBS to stop the trypsin reaction, the cells were collected into a test tube, and the cells were pelleted by centrifugation at 800 rpm for 3 minutes in a centrifuge. The cells were suspended in fresh MEMα medium containing 10% FBS in a manner of 4×10 5 cells/mL, and seeded in a 96-well plate at 100 μL/well, at 5 vol% CO 2 , 37°C, and humidified conditions Pre-culture for 1 day. (II): Sodium ascorbate and β-glycerophosphate disodium were dissolved in MEMα medium containing 10% FBS at 50 μg/mL and 10 mM, respectively, to prepare a differentiation induction medium for inducing differentiation into bone tissue. The test substance of Example 1 was dissolved in DMSO so that the concentration of the total amount of TGE and TGE glycosides was 20 mM, and it was diluted 200-fold with a differentiation induction medium, and then it was made into a specific concentration of 2. In a multi-fold manner, the MEMα medium containing 0.5% DMSO and 10% FBS was diluted to prepare a differentiation induction medium containing the test substance. The so-called specific concentration here means that the concentration of the total amount of TGE and TGE glycosides is 100 μM. (III): The culture medium of the pre-cultured cells in (I) will be removed, and the differentiation induction medium containing the test substance prepared in (II) will be added at 100 μl/well, and the medium will be replaced every 2 or 3 days. The cells were further cultured for 14 days under 5 vol% CO 2 , 37° C., and humid conditions. On the other hand, as a control group, a differentiation induction medium containing no test substance was added to other wells in the same amount, and the culture was carried out in the same manner. (IV) After culturing in the above (III), the medium was removed, and after washing once with serum-free DMEM, a Cell Counting-Kit 8 (Cell Counting-Kit 8) (Tongren Chemical) diluted to 30 times with serum-free DMEM medium was added. Company) 150 μL, incubate at 37°C until appropriate color development. The absorbance (A1) at 450 nm was measured using a plate reader (manufactured by Thermo Scientific). (V) Removal of Cell Counting Kit-8 from Medium After Assay of (IV), according to TRAP/ALP (tartrate-resistant acid phosphatase/alkaline phosphatase, tartrate-resistant acid phosphatase/alkaline phosphatase) staining kit (and The manual for the measurement of alkaline phosphatase (ALP) activity was carried out. Next, glycine, anhydrous magnesium chloride, and zinc chloride were dissolved in ultrapure water at 0.1 M, 1 mM, and 1 mM, respectively, and the pH was adjusted to 10.4 with sodium hydroxide to prepare a glycine buffer. liquid. One tablet of 4-nitrophenylphosphate disodium salt hexahydrate 5 mg (manufactured by Sigma Aldrich) was dissolved in 5 ml of the obtained glycine buffer. The resulting liquid was added to 150 μL of each well after ALP staining. After heating at 37° C. for 30 minutes, 100 μL of the supernatant was transferred to an assay plate (manufactured by AGC Techno Glass), and the absorbance (A2) at 405 nm was measured with a plate reader. The ALP activity was calculated|required by the following formula using the absorbance A1 and A2 measured above. The relative value of the ALP activity value obtained by ALP activity=A2/A1 was calculated by taking the control group (Comparative Example 1) as 100%. The results are shown in FIG. 2 . In general, ALP activity is widely used as a marker for inducing differentiation of MC3T3-E1 cells into osteoblasts. As shown in Figure 2, by adding the composition of Example 1 with a specific TGE:TGE glycoside amount ratio to MC3T3-E1 cells in differentiation induction medium, ALP activity was increased. This indicates that in MC3T3-E1 cells to which the composition of Example 1 was added, osteogenic differentiation was promoted. [Examples 2-1 to 2-8, Comparative Example 2] As test substances, those obtained by preparing TD, TGXG and TGE as described in Table 1 below were used. The TGE glycosides in Table 1 refer to the combined amount of TD and TGXG. The test substances of these Examples and Comparative Examples were used in the following sequence (i) to (viii) (lipolytic enzyme expression test). (In the table, numbers indicate mass ratios) (Lipolytic enzyme expression test) The gene expression of hormone-sensitive lipase (HSL) was measured by the following method. (i) Mouse fibroblast cells (3T3-L1) for differentiation into adipocytes were cultured in a preculture medium using a 75 cm 2 flask in a 37° C., 5 vol% CO 2 incubator. As a pre-culture medium, 10% FBS-DMEM was used. (ii) The trypsinized cells were suspended in 10% FBS-DMEM, and seeded into each well of a 96-well plate at a cell density of 2×10 3 cells/well. The seeded cells were pre-cultured for 2 days in a 37°C, 5 vol% CO 2 incubator. (iii) The medium was removed, and 3-Isobutyl-1-methylxanthine (3-Isobutyl-1-methylxanthine), 20 μg containing 1 μM dexamethasone, 1 mM 3-Isobutyl-1-methylxanthine and 20 μg were added at 100 μL/well. 10% FBS-DMEM (differentiation induction medium to adipocytes) prepared by the method of insulin/mL, and cultured for 2 days at 37° C. in a 5 vol% CO 2 incubator. (iv) Half of the medium was removed from each well, replaced with 10% FBS-DMEM (adipocyte differentiation maintenance medium) prepared in advance containing 20 μg/mL of insulin, and then incubated at 37°C in a 5 vol% CO 2 incubator cultured for 5 days. During this period, half of the medium was replaced with 10% FBS-DMEM containing insulin every 2 days. (v) The medium was completely removed from each well, and 100 μL/well (the total concentration of TGE and TGE glycosides was 100 μM) was added with 10% FBS-DMEM prepared in advance so that the test substance would be 100 μg/mL , and further cultured in a 37°C, 5 vol% CO 2 incubator for 48 hours. (vi) The medium was completely removed, washed twice with ice-cooled DPBS, and then ISOGEN (manufactured by NIPPON GENE) was added at 100 μL/well to purify total RNA according to the instruction manual. (vii) Using the ReverTra AceR qPCR RT Master Mix with gDNA Remover, according to the instruction manual, synthesize cDNA from the total RNA obtained in (vi). (viii) RT-PCR (Reverse Transcription-Polymerase Chain Reaction) was performed using the cDNA obtained in (vii) as a template using QuantiNova SYBR Green PCR kit. As a primer, Mm_Lipe_1_SG QuantiTect Primer Assay for HSL gene expression was used. Internal standards use Mm_Actb_1_SG QuantiTect Primer Assay for Actb. Regarding the expression level of the HSL gene, the relative values of Examples 2-1 to 2-8 when the gene expression level of Comparative Example 2 (control group) was set to 1 are shown in FIG. 3 . As shown in FIG. 3 , by using the test substances of each example, the gene expression level of hormone-sensitive lipase (HSL) was significantly higher than that of Comparative Example 2. Hormone-sensitive lipase (HSL) is known to be a neutral lipolytic enzyme and is present in adipocytes, decomposes triglycerides into free fatty acids and glycerol, and releases free fatty acids into the blood. Therefore, it was found that the composition of the present invention effectively promotes lipolysis in adipocytes by promoting the expression of hormone-sensitive lipase (HSL) gene. (Liver function improvement test) [Examples 3-1 to 3-16 and Comparative Example 3] (1) Cells were cultured in a 37°C, 5 vol% CO 2 incubator, using a 75 cm 2 flask, with 10% Human hepatoma-derived cells (HepG2, manufactured by ATCC Corporation) were cultured in a medium of fetal bovine serum (FBS). Next, the cells floated by trypsin treatment were seeded into each well of a 96-well plate from a 75 cm 2 flask at a cell density of 4.0×10 4 cells/well. After that, it was pre-incubated for 24 hours in a 37°C, 5 vol% CO 2 incubator. After removing the medium from each well, a medium containing the test substance prepared at a specific concentration was added at 100 μL/well, and cultured in a CO 2 incubator for 24 hours. As the medium in the medium containing the test substance, serum-free DMEM was used. As the test substance, those prepared by blending TD, TGXG, and TGE as described in Table 2 below were used. The TGE glycosides in Table 2 refer to the combined amount of TD and TGXG. As the above-mentioned specific concentration, the concentration of the total amount of TD, TGXG and TGE was 400 μg/ml. Comparative Example 3 was a control, and serum-free DMEM containing 0.5% DMSO was used to replace the medium containing the test substance. <Cell activity measurement> After the above-mentioned 24-hour culture, the medium was removed, and 150 μl per well of Cell Counting Kit-8 solution (manufactured by Dojin Chemical) diluted to 30 times by volume with serum-free DMEM was added to each well. After standing in a 37°C, 5 vol% CO 2 incubator for moderate color development, the absorbance at 450 nm was measured. Based on the obtained data, the % of control was calculated. The results obtained from the percentage of control = (Data sample - Data blank)/(Data control - Data blank) x 100 shown in Figure 4. The mean and standard deviation are shown in FIG. 4 . From the results shown in FIG. 4 , it can be seen that the amount of TGE and its glycosides has an excellent effect on protecting the activity of hepatocytes cultured in serum-free DMEM compared with specific irisin. Therefore, by using irisin with a specific mass ratio, it is possible to protect liver cells and improve liver function. [Example 4] As the test substance of Example 4, 43.14 mg of a powdered composition with a mass ratio of TGE:TGE glycoside=1:22.19 (TD:TGXG=1:1.81) (the total amount of TGE and TGE glycoside) was used ). This was used for the following cosmetic tests. (Cosmetic test) Two healthy and normal women (average age 19.7 years old) were used as subjects. The subjects were asked to ingest the test substance of Example 4 dissolved in water every morning. Keep it going for 12 weeks. Before the start of ingestion and after the start of ingestion (after 4, 8, and 12 weeks), the moisture content and evapotranspiration of the stratum corneum of the left cheek (near the center when the outer corner of the eye and the alar of the nose are connected with a straight line) were measured using the following measuring devices, respectively. , elasticity. The average value of the moisture content of the stratum corneum is shown in FIG. 5 , the average value of the water evapotranspiration amount is shown in FIG. 6 , and the average value of the elastic force is shown in FIG. 7 . • Cuticle moisture content: Measured using an epidermis cuticle moisture content measuring apparatus SKICON-200EX (manufactured by IBS Corporation). This apparatus for measuring the moisture content of the epidermis stratum corneum evaluates the skin conductivity (conductance, unit: μS) as the moisture content of the stratum corneum. • The amount of water evapotranspiration: measured by Tewameter TH300 (manufactured by Courage+Khazaka Corporation). • Elasticity: Measured with a skin viscoelasticity measuring device Cutometer MPA580 (manufactured by Courage + Khazaka Corporation). Regarding the elasticity, the recovery elasticity ratio is used as a measurement parameter of the skin viscoelasticity value obtained by the Cutometer as a measuring device. The displacement amount of the skin restored by the vertex is analyzed and obtained. The closer the recovery elasticity ratio is to 1.00, the higher the elasticity of the skin. These measurements were performed using standard methods described in the instructions included with the machine. From the results of FIGS. 5 to 7 , it was found that as the ingestion time of the composition of the present invention with a specific TGE:TGE glycoside ratio increased, the moisture content of the stratum corneum of the skin was increased, the amount of water evapotranspiration was suppressed, and the elasticity was improved. The following formulation examples of the composition of the present invention are listed, but the following formulation examples do not limit the present invention. Formulation Examples 1 to 10: Tablets Each component was mixed so that the ratios in Table 3 were obtained, and tableting was performed according to the usual practice to manufacture the lozenges (250 mg per tablet) of Formulation Examples 1 to 10. The tablets of Preparation Examples 1 to 10 are excellent in that the effects of the present invention, such as an increase in the moisture content of the stratum corneum, improvement of skin elasticity, and inhibition of skin moisture transpiration, are particularly well exhibited. [table 3] Formulation Examples 11 to 20: Soft Capsules Soft capsules (1 capsule of 300 mg) were prepared by preparing the content of the soft capsules in a proportion as shown in Table 4 and filling them in the capsule film. The capsule membrane liquid is casted to form a film, and the content liquid is filled in the interior for heat sealing, and the formed soft capsule is dried to form a capsule. The soft capsules of Preparation Examples 11 to 20 are excellent in that the effects of the present invention, such as increasing the moisture content of the stratum corneum, improving skin elasticity, and inhibiting skin moisture transpiration, are particularly well exhibited. Formulation Examples 21 to 30: Powder (granules) According to Table 5, each component was put into a Flow Coater NFLO-200 type fluid bed granulator (manufactured by Freund Corporation), and mixed for several minutes under airflow. Granulation is carried out by spraying it with water. Next, the obtained granules were screened with a 30-mesh sieve to produce granules. The granules of Preparation Examples 21 to 22 are excellent in that the effects of the present invention, such as increasing the moisture content of the stratum corneum, improving skin elasticity, and inhibiting skin moisture transpiration, are particularly well exhibited. Preparation Examples 21-22: Liquid preparations (liquid beverages) Each component was prepared in the proportions shown in Table 6 to prepare liquid beverages. The liquid beverages of Preparation Examples 31 to 40 are excellent in that the effects of the present invention, such as increasing the moisture content of the stratum corneum, improving skin elasticity, and inhibiting transpiration of skin moisture, are particularly well exhibited.
