KR20190079035A - Composition for promoting hair growth comprising extract of Houttuynia cordata as effective component - Google Patents
Composition for promoting hair growth comprising extract of Houttuynia cordata as effective component Download PDFInfo
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- KR20190079035A KR20190079035A KR1020170180934A KR20170180934A KR20190079035A KR 20190079035 A KR20190079035 A KR 20190079035A KR 1020170180934 A KR1020170180934 A KR 1020170180934A KR 20170180934 A KR20170180934 A KR 20170180934A KR 20190079035 A KR20190079035 A KR 20190079035A
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- extract
- hair
- hair growth
- composition
- acid
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Abstract
Description
본 발명은 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 조성물에 관한 것이다.The present invention relates to a composition for promoting hair growth comprising an extract of Rhododendron japonica as an active ingredient.
인체의 모발은 평균적으로 10만 개 정도이며, 이들은 성장기(anagen), 퇴행기(catagen), 휴지기(telogen), 탈모기(exogen)의 주기적인 변화를 거치면서 생성, 소멸을 반복한다. 탈모란 모발 주기의 성장기에 해당하는 기간이 여러 원인으로 인해 짧아져 퇴행기와 휴지기 모발의 비율이 많아지는 것을 말하며, 탈모의 원인은 개인에 따라 다르지만, 유전적인 요인, 노화에 의한 두피 세포의 퇴화, 모근에 공급되는 영양소의 부족, 두피 지방질 과다로 인한 산소공급 부족, 지방질이나 이물질 또는 두피 질환에 의한 염증과 모공 폐쇄 현상, 스트레스 등이 있다. 또한, 혈액순환이 잘 되면 모근에 영양분과 산소가 원활하게 공급되어 모발이 건강하게 자랄 수 있지만, 고지방식이나 흡연 등으로 혈액순환이 원활하지 않으면 탈모가 촉진될 수 있다. The human body has an average of about 100,000 hairs, and they repeatedly produce and disappear through periodic changes of anagen, catagen, telogen, and exogen. Hair loss refers to the fact that the period corresponding to the growth period of the hair cycle is shortened due to various causes, and the ratio of the retrogressive hair to the resting hair increases. The cause of hair loss varies depending on the individual, but genetic factors, Lack of nutrients supplied to the hair follicles, lack of oxygen supply due to excessive amount of scalp, inflammation due to lipid or foreign matter or scalp disease, clogging of the pores, and stress. In addition, if the blood circulation is good, nutrients and oxygen are supplied smoothly to the hair follicles and the hair can grow healthily. However, hair loss can be promoted if the blood circulation is not smooth due to the high fat diet or smoking.
유전적 요인에 의한 탈모에는 남성호르몬이 관여한다. 남성호르몬의 하나인 테스토스테론(testosterone)은 5알파-리덕타아제(5α-reductase)라는 효소에 의해서 활성형인 디하이드로테스토스테론(dihydrotestosterone)으로 전환된다. 사춘기 이후 과량 분비된 디하이드로테스토스테론은 피지세포(sebocyte)에서 피지를 과잉생성하는 주된 원인 물질로 작용하여 여드름 및 남성형 탈모를 유발한다. 특히 모낭 세포에서도 테스토스테론에 비하여 디하이드로테스토스테론이 호르몬 수용체에 더욱 잘 결합하기 때문에 탈모를 일으키는 원인을 제공한다.Male genitals are involved in hair loss caused by genetic factors. Testosterone, one of the male hormones, is converted to the active dihydrotestosterone by an enzyme called 5α-reductase. After puberty, excess secreted dihydrotestosterone acts as a major cause of excess sebum production in sebocytes, leading to acne and male hair loss. In particular, hair follicle cells are more likely to cause hair loss because dihydrotestosterone binds more to the hormone receptor than testosterone.
또한, 두피의 기저부에서는 끊임없이 세포분열이 일어나 점차 피부표면으로 밀려가면서 각화되어 두피의 제일 바깥쪽에 각질층을 형성하게 되는데, 이는 시간이 지나면 표피가 떨어져 나가 비듬을 형성하게 된다. 이 비듬이 피지선에서 나오는 피지와 혼합하여 지루가 되는데, 이것이 모공을 막아 영양장애와 위축작용을 일으키면서 머리카락이 빠지게 된다. 지루 자체 역시 남성 호르몬 작용에 의한 이차적인 현상으로, 지루 자체가 탈모의 원인은 아니다. 즉 남성호르몬은 머리카락을 가늘게 하지만 피지선을 비대시켜 피지의 분비를 증가시킨다. 피지선이 비대해지고 기능이 활발해지면, 머리의 표면에 잔비듬이 많아지고 5알파-리덕타아제의 분비도 늘어나 모발의 발육과 성장에 영향을 주게 된다.In addition, at the base of the scalp, cell division occurs constantly, and it is gradually crooked to the surface of the skin and becomes keratinized to form a stratum corneum on the outermost side of the scalp. This dandruff mixes with the sebum coming out of the sebaceous glands and becomes bored, which blocks the pores and causes nutritional disorders and atrophy, resulting in hair loss. Boring itself is a secondary phenomenon due to the action of male hormones, and boring itself is not the cause of hair loss. In other words, male hormones thin hair, but the sebaceous glands increase the secretion of sebum. When the sebaceous gland becomes enlarged and functioning, the dandruff on the surface of the hair increases and the secretion of 5-alpha-reductase increases, affecting hair growth and growth.
탈모는 원인이 다양한 만큼 이를 해결하기 위한 방법도 다양하다. 탈모를 방지하고 육모를 촉진하기 위해서 혈관을 확장시켜 모근에 영양 공급이 잘 이루어지도록 하거나, 모발의 구성 성분과 유사한 성분을 직접 모근에 공급하는 외용제나 건강보조 식품, 의약품 등이 출시되고 있다. 하지만, 화학약품의 사용으로 인하여 약물이상 반응(협심증, 피부병, 가려움증 등)이나 부작용에 대한 사례가 보고되어 있으며, 탈모 치료를 위해 고가의 돈을 투자하지만 그만큼의 효과를 얻지 못하는 등의 문제점이 있다.Hair loss can be as diverse as the variety of causes. To prevent hair loss and promote hair growth, blood vessels are expanded to supply nutrients to the hair follicles, and external preparations, health supplements, and medicines that directly supply the hair root with components similar to those of the hair are being marketed. However, there have been reported cases of adverse drug reactions (angina, skin diseases, itching, etc.) and side effects due to the use of chemicals, and there is a problem that expensive money is invested for treating hair loss, .
어성초(Houttuynia cordata)는 삼백초과(Saururaceae)의 식물인 약모밀의 전초로, 아시아 동남부와 특히 일본, 한국 등지에 서식한다. 상기 어성초는 열가지 약효가 있다고 하여 십약이라고도 부르며, 줄기는 고구마 잎과 같고 생잎을 만지면 생선 비린내가 심하게 나는 것에서 유래되어 어성초로 부르기도 한다. 어성초는 약용 및 식용으로도 가능하며 식품공전 식품 원재료 분류에 부원료로 최소량만을 사용할 수 있는 동ㆍ식물로 분류되어 있고, 약리적으로는 강심, 이뇨, 항균, 해독, 항암 효능이 있는 것으로 널리 알려져 있으며, 민간에서는 해독과 미용을 돕는 화장품 및 건강기능식품으로 이용하고 있다. Houttuynia cordata is an outbreak of anomalies, a plant of the Saururaceae, which lives in southeastern Asia and especially in Japan and Korea. It is said that there are ten kinds of phytophthora, and the stem is like leaf of sweet potato. It can be used for medicinal and edible purposes. It is classified as a plant or a plant which can only use the minimum amount as a supplementary ingredient in food raw material classification. It is widely known to have pharmacological effects such as strength, diuretic, antimicrobial, In the private sector, they are used as cosmetics and health functional foods to help detoxification and beauty.
한편, 한국등록특허 제1661245호에는 '호의초, 황련, 어성초, 인삼, 작약, 모란, 효모액기스, 하수오를 함유하는 피부 및 두피보호, 탈모방지용 한방 화장료 조성물 및 그 제조방법'이 개시되어 있고, 한국공개특허 제2016-0109332호에는 '가열처리 인삼, 어성초, 자소엽, 및 녹차엽으로 구성된 조합 추출물을 유효성분으로 함유하는 탈모 방지 및 발모 촉진 효과를 갖는 조성물'이 개시되어 있으나 본 발명의 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 조성물에 대해서는 기재된 바가 없다.On the other hand, Korean Patent No. 1661245 discloses a herbal cosmetic composition for protecting skin and scalp, preventing hair loss, and a method for producing the same, which comprises a composition comprising a compound of formula (I), a compound of formula (I), a compound of formula Korean Patent Laid-Open Publication No. 2016-0109332 discloses a composition having an effect of preventing hair loss and promoting hair growth, which comprises a combination extract comprising a heat-treated ginseng, a zelkova, a lobule, and a green tea leaf as an active ingredient. There is no description about a composition for promoting hair growth that contains an extract as an active ingredient.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명자들은 어성초 추출물이 모유두세포에는 독성이 없으며, 지방전구 세포를 분화시키고, 항산화 활성이 우수한 것을 확인함으로써, 본 발명을 완성하였다.The present invention has been made in view of the above-mentioned needs. The present inventors have completed the present invention by confirming that Hwasungcho extract has no toxicity to dermal papilla cells, differentiates lipid precursor cells, and has excellent antioxidative activity.
상기 과제를 해결하기 위해, 본 발명은 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 화장료 조성물, 건강기능식품 조성물 및 약학 조성물을 제공한다.In order to solve the above problems, the present invention provides a cosmetic composition for accelerating hair growth, a health functional food composition, and a pharmaceutical composition, which comprise an extract of Rhododendron japonica as an active ingredient.
본 발명에 따른 어성초 추출물은 세포독성이 거의 없고 다양한 산화적 스트레스로부터 신체를 보호할 수 있는 항산화 효과가 우수하므로, 탈모 방지 또는 육모 촉진을 위한 화장료, 건강기능식품 또는 약학 조성물의 신규한 천연 소재로 유용하게 사용될 수 있다.Since the extract of Hwasung extract according to the present invention has little cytotoxicity and is excellent in antioxidative effect for protecting the body from various oxidative stresses, it can be used as a novel natural material for cosmetics, health functional food or pharmaceutical composition for preventing hair loss or promoting hair growth Can be usefully used.
도 1은 어성초 에탄올 추출물(A)과, 어성초 에탄올 추출물 및 5개 분획물(B, Hexane; 헥산, CHCl3; 클로로포름, E.A; 에틸아세테이트, n-butanol; 부탄올, DW; 증류수)의 TLC 분석 결과이다. R f 0.50; 클로로겐산(chlorogenic acid), R f 0.54; 하이퍼로사이드(hyperoside), R f 0.79; 퀘르시트린(quercitrin), 및 R f 0.88; 아프젤린(afzelin).
도 2는 표준시약(A~C) 및 어성초 에탄올 추출물(D~F)의 HPLC 결과이다. A와 D는 280 nm; B와 E는 320 nm; 및 C와 F는 280 nm에서 확인한 것이다. 1; 갈산, 2; 쿠마르산, 3; 살리실산, 4; 트랜스-신남산, 5; 퀘르시트린, 6; 쿼세틴, 7; 클로로겐산, 8; 이소퀘르시트린, 9; 하이퍼로사이드.
도 3은 섬유아세포에서 어성초 에탄올 추출물의 세포독성을 분석한 결과이다.
도 4는 인간 모유두세포에서 어성초 에탄올 추출물의 세포독성을 분석한 결과이다.
도 5는 지방전구세포에서 어성초 에탄올 추출물의 세포독성을 분석한 결과이다.
도 6은 어성초 에탄올 추출물의 처리에 따른 지방전구세포(3T3-L1)에서 지방세포로의 분화율(A)과 Oil-red O 염색(B) 결과이다. DMⅡ; 분화유도 물질.
도 7은 어성초 에탄올 추출물의 항산화 효과를 분석하기 위해 전자공여능(A), ABTS 라이칼 소거능(B) 및 수퍼옥사이드 음이온 소거능(C)을 측정하여 나타낸 그래프이다. Vitamin C; 대조군.1 shows the result of TLC analysis of a water extract of a water-soluble herbicide (A), a water extract of a herb of a plant and five fractions (B, hexane, hexane, CHCl 3 ; chloroform, EA; ethyl acetate, n-butanol, butanol, DW; distilled water) . R f 0.50; Chlorogenic acid, R f 0.54; Hyperoside, R f 0.79; Quercitrin, and R f 0.88; Afzelin.
2 shows the HPLC results of standard reagents (A to C) and Hwasungcho ethanol extracts (D to F). A and D are 280 nm; B and E are 320 nm; And C and F were confirmed at 280 nm. One; Gallic acid, 2; Cumaric acid, 3; Salicylic acid, 4; Trans-cinnamic acid, 5; Quercitrin, 6; Quercetin, 7; Chlorogenic acid, 8; Isoquercitrin, 9; Hyde to the side.
FIG. 3 shows the results of analysis of cytotoxicity of the extract of Hwasungcho extract in fibroblasts.
Fig. 4 shows the result of analysis of cytotoxicity of Hwasungcho ethanol extract in human dermal papilla cells.
FIG. 5 shows the results of cytotoxicity analysis of the ethanol extract of Hwasungcho extract from lipid precursor cells.
FIG. 6 shows the results of differentiation (A) and oil-red O staining (B) from adipocyte precursor cells (3T3-L1) DM II; Differentiation inducing substance.
7 is a graph showing electron donating ability (A), ABTS radical scavenging ability (B), and superoxide anion scavenging ability (C) measured for analyzing the antioxidative effect of ethanol extract of Hwasungcho. Vitamin C; Control group.
본 발명의 목적을 달성하기 위하여, 본 발명은 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 화장료 조성물을 제공한다.In order to accomplish the object of the present invention, the present invention provides a cosmetic composition for accelerating hair growth comprising an extract of Aspergillus as an active ingredient.
본 발명의 일 구현 예에 따른 육모 촉진용 화장료 조성물에 있어서, 상기 어성초 추출물은 물, 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합 용매를 추출용매로 이용한 것일 수 있으며, 바람직하게는 탄소수 1 내지 4의 저급 알코올을 이용한 추출물일 수 있고, 더욱 바람직하게는 에탄올 추출물일 수 있으나, 이에 제한되지 않는다.In the cosmetic composition for promoting hair growth in accordance with an embodiment of the present invention, the extract may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, as an extraction solvent, preferably 1 to 4 carbon atoms Of lower alcohol, and more preferably ethanol extract, but is not limited thereto.
또한, 본 발명의 일 구현 예에 따른 육모 촉진용 화장료 조성물에서, 상기 어성초 추출물의 제조방법은 구체적으로는,Further, in the cosmetic composition for promoting hair growth according to an embodiment of the present invention,
1) 어성초를 150~350 메쉬가 되도록 분쇄하는 단계;1) pulverizing the seedlings to a size of 150 to 350 mesh;
2) 상기 단계 1)의 어성초 분말 1 중량부에 대하여 30~50 중량부의 에탄올을 각각 첨가하여 24~60시간 동안 추출하는 단계; 및2) adding 30 to 50 parts by weight of ethanol to 1 part by weight of the seedling powder of step 1), and extracting the mixture for 24 to 60 hours; And
3) 상기 단계 2)의 어성초 추출물을 여과하는 단계;를 포함할 수 있고, 더욱 구체적으로는, 3) filtering the Horsetail extract of step 2), and more specifically,
1) 어성초를 200~300 메쉬가 되도록 분쇄하는 단계; 1) pulverizing the seeds so as to have a mesh of 200 to 300 mesh;
2) 상기 단계 1)에서 분쇄된 어성초 분말 1 중량부에 대하여 40 중량부의 에탄올을 각각 첨가하여 48시간 동안 추출하는 단계; 및2) adding 40 parts by weight of ethanol to 1 part by weight of crushed asps powder in step 1), and extracting the mixture for 48 hours; And
3) 상기 단계 2)의 어성초 추출물을 여과하는 단계;를 포함할 수 있으나, 이에 제한되지 않는다.3) filtering the Horsetail extract of step 2), but is not limited thereto.
또한, 본 발명의 일 구현 예에 따른 육모 촉진용 화장료 조성물에 있어서, 상기 3) 단계에서 획득한 추출물은 상기 단계 2)를 2회 더 반복하여 획득한 추출물일 수 있으며, 바람직하게는 상기 단계 2)에서 여과한 추출물과, 여과하고 남은 잔여물에 에탄올을 첨가하여 추출하고 여과하는 단계를 2회 더 반복하여 획득한 추출물을 혼합한 추출물일 수 있으나, 이에 제한되지 않는다.In the cosmetic composition for promoting hair growth according to an embodiment of the present invention, the extract obtained in the step 3) may be an extract obtained by repeating the step 2) twice more preferably the step 2 ), And an extract obtained by adding ethanol to the remaining residue after filtration and extracting and filtering the extract twice. The extract is not limited thereto.
본 명세서에서 용어 "육모 촉진"은 모발의 생성 및 성장을 촉진하여 궁극적으로 전체 모발에서 성장기 모발의 비중을 증가시키는 것을 의미한다. 따라서, 상기 용어는 성장기 모발의 비중이 감소함으로써 유발되는 탈모를 억제하는 것을 의미하므로, "탈모 개선", "탈모 예방" 및 "탈모 치료"와 동일한 의미를 가진다.As used herein, the term " hair growth promoting "refers to promoting the production and growth of hair, ultimately increasing the specific gravity of growing hair in total hair. Therefore, the term has the same meaning as "hair loss improvement "," hair loss prevention ", and "hair loss treatment" since it means suppressing hair loss caused by a decrease in specific gravity of growing hair.
또한, 본 발명의 일 구현 예에 따른 육모 촉진용 화장료 조성물에 있어서, 상기 조성물은 헤어 컨디셔너, 헤어 토닉, 헤어 크림, 헤어 로션, 헤어 오일, 헤어 스프레이, 샴푸 또는 린스의 제형으로 제조되는 것일 수 있으나, 이에 제한되지 않는다.In the cosmetic composition for promoting hair growth according to an embodiment of the present invention, the composition may be a hair conditioner, a hair tonic, a hair cream, a hair lotion, a hair oil, a hair spray, a shampoo or a rinse, , But is not limited thereto.
상기 화장료 조성물은 화장료 조성물에 통상적으로 이용되는 성분들을 포함하며, 예를 들어 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다.The cosmetic composition includes components commonly used in cosmetic compositions and includes, for example, customary adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers.
상기 담체는 정제수, 일가 알코올류(에탄올 또는 프로필 알코올), 다가 알코올류(글리세롤, 1,3-부티렌글리콜 또는 프로필렌글리콜), 고급 지방산류(팔미틸산 또는 리놀렌산), 유지류(소맥 배아유, 동백기름, 호호바유, 올리브유, 스쿠알렌, 해바라기유, 마카데미아땅콩유, 아보가드유, 대두 수첨가 레시틴 또는 지방산 글리세라이드) 등을 사용할 수 있으나, 이에 한정되지 않는다. 또한, 필요에 따라 계면활성제, 살균제, 산화방지제, 자외선 흡수제, 소염제 및 청량제를 첨가할 수 있다.The carrier may be selected from the group consisting of purified water, monohydric alcohols (ethanol or propyl alcohol), polyhydric alcohols (glycerol, 1,3-butylene glycol or propylene glycol), higher fatty acids (palmitic acid or linolenic acid) But are not limited to, oil, jojoba oil, olive oil, squalane, sunflower oil, macadamia peanut oil, avocado oil, soybean water-added lecithin or fatty acid glyceride). If necessary, a surfactant, a bactericide, an antioxidant, an ultraviolet absorber, an anti-inflammatory agent and a refreshing agent may be added.
상기 계면활성제는 폴리옥시에틸렌, 경화 피마자유, 폴리옥시에틸렌, 올레일에테르, 모노올레인산폴리옥시에틸렌, 폴리옥시에틸렌, 글리세릴모노스테아레이트, 모노스테아린산소르비탄, 모노올레인산폴리옥시에틸렌, 소르비탄, 자당지방산에스테르, 모노라우린산헥사글리세린, 폴리옥시에틸렌 환원라놀린, POE, 글리세릴피로글루타민산, 이소스테아린산, 디에스테르, N-아세틸글루타민 및 이소스테아릴에스테르로 이루어진 군에서 선택적으로 포함할 수 있다.The surfactant is selected from the group consisting of polyoxyethylene, hardened castor oil, polyoxyethylene, oleyl ether, polyoxyethylene monooleate, polyoxyethylene, glyceryl monostearate, monostearic acid sorbitan, monooleic acid polyoxyethylene, sorbitan, Polyglycerol fatty acid ester, sucrose fatty acid ester, monolauric acid hexaglycerin, polyoxyethylene reduced lanolin, POE, glyceryl pyroglutamic acid, isostearic acid, diester, N-acetylglutamine and isostearyl ester.
상기 살균제는 히녹티올, 트리크로산, 크롤헥시딘글루콘산염, 페녹시에탄올, 레조르신, 이소프로필메틸페놀, 아즐렌(azulene),살리실산 및 징크피리타온으로 이루어진 군에서 선택적으로 포함할 수 있다.The fungicide may optionally be selected from the group consisting of: hynochitol, trichloro acid, chlorohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid and zinc pyrithione have.
상기 산화방지제는 부틸히드록시아니솔, 몰식자산, 몰식자산프로필 및 에리소르빈산 중에서 어떠한 것도 사용가능하다.As the antioxidant, any of butylhydroxyanisole, gallic acid, propyl gallate, and erythorbic acid may be used.
상기 자외선 흡수제는 디히드록시벤조페논 등의 벤조페논류, 멜라닌, 파라아미노벤조산에틸, 파라디메틸아미노벤조산 2-에틸헥실에스테르, 시녹사이트, 파라메톡시계피산 2-에틸헥실에스테르, 2-(2-히드록시-5-메틸페닐) 벤조트리아졸, 우로카닌산 및 금속산화물 미립자 중에서 어떠한 것도 사용가능하다.The ultraviolet absorber may be at least one selected from the group consisting of benzophenones such as dihydroxybenzophenone and the like, melanin, ethyl paraaminobenzoate, 2-ethylhexyl paradimethylbenzoate, synonym, 2-ethylhexyl phamethoxycinnamate, 2- Hydroxy-5-methylphenyl) benzotriazole, urocanic acid, and metal oxide fine particles.
상기 소염제로는 글리틸리틴산디칼륨 또는 알란토인을 사용할 수 있고, 청량제로는 고추틴크 또는 1-멘톨을 사용할 수 있다.As the anti-inflammatory agent, dipotassium glycyrrhizinate or allantoin can be used, and as a refreshing agent, pepper tincture or 1-menthol can be used.
또한, 본 발명에 따른 화장료 조성물은 어성초 추출물 이외에, 본 발명의 작용(육모 작용)을 손상시키지 않는 한도에서 종래부터 사용되어오던 양모제 또는 육모제를 함께 혼합하여 사용할 수 있다.Further, the cosmetic composition according to the present invention can be used in combination with a hair or hair restorer conventionally used as long as the action (hair growth) of the present invention is not impaired.
본 발명은 또한, 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 건강기능식품 조성물을 제공한다.The present invention also provides a health functional food composition for accelerating hair growth comprising an extract of Rhododendron japonica as an active ingredient.
상기 건강기능식품 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽, 발포정 및 음료 중에서 선택된 어느 하나의 제형으로 제조될 수 있으나, 이에 제한되지 않는다. 상기 건강기능식품 조성물은 육모를 촉진하기 위해 섭취할 수 있는 것이면 특별히 제한되지 않는다.The health functional food composition may be prepared in any form of powder, granule, ring, tablet, capsule, candy, syrup, foamable beverage and beverage, but is not limited thereto. The health functional food composition is not particularly limited as long as it can be ingested to promote hair growth.
본 발명의 어성초 추출물을 식품첨가물로 사용하는 경우, 상기 어성초 추출물을 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분은 그의 사용 목적(예방 또는 개선)에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 어성초 추출물의 발효물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양의로 첨가된다. 그러나 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 안전성 면에서 아무런 문제가 없는 범위의 양으로 사용될 수 있다.When the Horticultural Extract of the present invention is used as a food additive, the Horticultural Extract may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The active ingredient may be suitably used depending on its intended use (prevention or improvement). Generally, the fermented product of the extract of Hwasung extract of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material. However, in the case of long-term intake for the purpose of controlling health, it can be used in an amount that does not cause any safety problems.
상기 식품의 종류에는 특별한 제한은 없다. 상기 건강기능식품 조성물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다. 상기 음료는 탄산음료, 기능성이온음료, 쥬스(예를 들어, 사과, 배,포도, 알로에, 감귤, 복숭아, 당근, 토마토쥬스 등), 식혜 등을 포함한다. There is no particular limitation on the kind of the food. Examples of the foods to which the health functional food composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense. The beverage includes carbonated beverages, functional ionic beverages, juice (e.g., apple, pear, grape, aloe, citrus, peach, carrot, tomato juice, etc.), sikhye, and the like.
본 발명의 기능성 식품은 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소 및 조미제를 포함한다. 예컨대, 드링크제로 제조되는 경우에는 유효성분 이외에 천연 탄수화물 또는 향미제를 추가 성분으로서 포함시킬 수 있다. 상기 천연 탄수화물은 모노사카라이드(예컨대, 글루코오스, 프럭토오스 등), 디사카라이드(예컨대, 말토스, 수크로오스 등), 올리고당, 폴리사카라이드(예컨대, 덱스트린, 시클로덱스트린 등) 또는 당알코올(예컨대, 자일리톨, 소르비톨, 에리쓰리톨 등)인 것이 바람직하다. 상기 향미제는 천연 향미제(예컨대, 타우마틴, 스테비아 추출물 등)와 합성 향미제(예컨대, 사카린, 아스파르탐 등)를 이용할 수 있다. 상기 건강기능식품 조성물 외에 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 더 함유할 수 있다.The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, in the case of a drink, a natural carbohydrate or a flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.). In addition to the above-mentioned health functional food composition, it is possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like.
본 발명은 또한, 어성초 추출물을 유효성분으로 함유하는 육모 촉진용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for accelerating hair growth comprising an extract of Aspergillus as an active ingredient.
본 발명의 상기 어성초 추출물을 포함하는 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 더 포함할 수 있다. The composition comprising the perennial herb extract of the present invention may further comprise an appropriate carrier, excipient or diluent conventionally used in the production of a pharmaceutical composition.
본 발명의 약학 조성물의 약학적 투여 형태는 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 조합으로 사용될 수 있다.The pharmaceutical dosage forms of the pharmaceutical compositions of the present invention may be used alone or in combination with other pharmaceutically active compounds as well as in suitable combinations.
본 발명에 따른 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제제, 외용제, 좌제 및 주사제의 형태로 제형화하여 사용될 수 있다.The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and injections according to conventional methods .
상기 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다.Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 발효물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트, 수크로오스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이 외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로골, 트윈 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
본 발명의 상기 약학 조성물의 바람직한 투여량은 환자의 상태 및 체중,질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 본 발명의 약학 조성물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내(intracerebroventricular) 주사에 의해 투여될 수 있다.
The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the administration route and the period of time, but can be appropriately selected by those skilled in the art. The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.
제조예 1. 어성초 추출물 제조Production Example 1. Preparation of Houttuynia cordata extract
어성초는 지리산 D&H 사로부터 구입하였으며, 어성초 건조물을 200~300 메쉬가 되도록 믹서기로 분쇄하여 사용하였다. 분쇄된 어성초 분말 50 g을 70%(v/v) 에탄올 2,000 mL에 넣고 48시간 동안 상온 추출하였다. 상기 어성초 추출물은 필터(3M사, 미국)로 여과하여 4℃에 보관하였고, 여과한 후 어성초 잔여물에 70%(v/v) 에탄올 500 mL을 넣고 혼합하여 24시간 동안 상온 추출한 후 추출물은 다시 필터로 여과하여 4℃에 보관하였고, 어성초 잔여물에 70%(v/v) 에탄올 500 mL을 넣고 혼합하여 24시간 동안 상온 추출한 뒤 추출물은 필터로 여과하여 4℃에 보관하였던 추출물과 혼합한 다음 40℃ 및 55 rpm의 조건으로 감압농축 후 동결건조하여 4℃에 보관하여 사용하였다.
Hwasungcho was purchased from Jiri Mountain D & H Co., and crushed by using a blender so as to have 200-300 mesh. 50 g of pulverized cord blood powder was added to 2,000 mL of 70% (v / v) ethanol and extracted at room temperature for 48 hours. After filtration, 500 mL of 70% (v / v) ethanol was added to the filtrate, and the mixture was incubated at room temperature for 24 hours. The mixture was filtered through a filter and stored at 4 ° C. To the supernatant, 500 mL of 70% (v / v) ethanol was added and mixed. After 24 hours of room temperature extraction, the extract was filtered and mixed with the extract Concentrated under reduced pressure at 40 ° C and 55 rpm, lyophilized and stored at 4 ° C.
실시예 1. 박층크로마토그래피(thin-layer chromatography, TLC) 분석Example 1. Thin-layer chromatography (TLC) analysis
어성초의 표준생약 규격 기준에 따라 어성초의 에탄올 추출물, 헥산 추출물, 클로로포름 추출물, 아세트산에틸 추출물 및 부탄올 추출물을 이용하여 박층크로마토그래피를 수행하였으며, 박층크로마토그래피의 전개용매로는 아세트산에틸(ethyl acetate):증류수(distilled water):포름산(formic acid):아세트산(acetic acid)이 100 : 15 : 7 : 7 비율로 혼합된 혼합용액을 사용하여 전개시켰다.Thin layer chromatography was performed using ethanol extract, hexane extract, chloroform extract, ethyl acetate extract and butanol extract of Rhododendron japonica according to the standards of standardized herbal medicine. The developing solvent for the thin layer chromatography was ethyl acetate: Distilled water: Formic acid: Acetic acid was developed using mixed solution of 100: 15: 7: 7 ratio.
UV/Vis 단파장(254 nm)과 장파장(365 nm) 및 10% 황산(sulfuric acid, H2SO4) 발색 결과, 어성초 에탄올 추출물에서 어성초의 4가지 주요성분인 클로로겐산(chlorogenic acid), 하이퍼로사이드(hyperoside), 퀘르시트린(quercitrin) 및 아프젤린(afzelin)이 포함되어 있음을 확인하였으며, 특히 퀘르시트린 성분이 뚜렷하게 검출됨을 확인하였다(도 1A). 상기 성분들은 참고문헌(한약재 품질표준화사업단 연구개발과제 최종보고서(어성초), 2014, 식품의약품안전처. Anal Sci. 2015. 31(6). 535-41)에 기재된 각 성분별 R f 값(각각 0.5, 0.54, 0.79, 0.88)과 비교하여 분석하였다. As a result of UV / Vis short wavelength (254 nm), long wave length (365 nm) and 10% sulfuric acid (H 2 SO 4 ) coloring, the four main components of Hwasungcho, chlorogenic acid, hyperoside, quercitrin and afzelin. Especially, it was confirmed that the quercitrin component was clearly detected (FIG. 1A). The above-mentioned components are the values of R f for each component described in the reference document (Analytical Sci. 2015, 31 (6), 535-41, 2014, 0.5, 0.54, 0.79, 0.88).
또한, 상기에서 제조된 어성초 에탄올 추출물에서 헥산(n-hexane) 분획물, 클로로포름(chloroform, CHCl3) 분획물, 아세트산에틸(ethyl acetate) 분획물, 부탄올(butanol) 분획물 및 증류수 분획물의 총 5개 분획물을 얻었으며, 상기 어성초 추출물과 5가지 분획물을 이용하여 박층크로마토그래피를 수행하였다. In addition, five fractions were obtained from the Horseshoe ethanol extract prepared above, namely, n-hexane fraction, chloroform (CHCl 3 ) fraction, ethyl acetate fraction, butanol fraction and distilled water fraction Thin layer chromatography was carried out using the above extracts and five fractions.
그 결과, 어성초 에탄올 추출물 및 5개의 분획물 중 아세트산에틸 분획물이 다른 분획물에 비해 주요성분들이 가장 많이 포함되어 있음을 확인하였다(도 1B).
As a result, it was confirmed that the ethyl acetate fraction and the ethyl acetate fraction of the five fractions contained the most major components than the other fractions (FIG. 1B).
실시예 2. HPLC(high pressure liquid chromatography) 분석Example 2. HPLC (high pressure liquid chromatography) analysis
표준시약과 어성초 에탄올 추출물에 대한 HPLC 분석을 통해 정량 평가를 수행하였으며, HPLC 분석 조건은 하기 표 1과 같다. 정량 평가에 사용된 페놀산(phenolic acid) 표준품은 클로로겐산(chlorogenic acid), 신남산(cinnamic acid), p-쿠마르산(coumaric acid), 갈산(gallic acid), 살리실산(salicylic acid) 등을 선정하였고, 플로보노이드(flavonoid) 표준품은 하이퍼로사이드(hyperoside), 쿼세틴(quercetin), 이소퀘르시트린(isoquercitrin), 퀘르시트린(quercitrin) 등을 선정하여 평가에 활용하였다. Quantitative evaluation was carried out by HPLC analysis of the standard reagent and Hwasungcho ethanol extract. HPLC analysis conditions are shown in Table 1 below. The phenolic acid standard used in the quantitative evaluation was selected from chlorogenic acid, cinnamic acid, coumaric acid, gallic acid, and salicylic acid. And flavonoid standards were selected for hyperoside, quercetin, isoquercitrin, quercitrin, and the like.
그 결과, 표준시약의 보유시간(retention time)은 약 28.216분에서 갈산(1); 약 31.823분에서 쿠마르산(2); 약 37.525분에서 살리실산(3); 약 40.006분에서 트랜스-신남산(4); 약 33.052분에서 퀘르시트린(5); 약 39.300분에서 쿼세틴(6); 약 26.078분에서 클로로겐산(7); 약 30.762분에서 이소퀘르시트린(8); 약 30.974분에서 하이퍼로사이드(9)에 해당하는 피크가 관찰되었다(도 2A~C).As a result, the retention time of the standard reagent was about 28.216 min (1); Cumaric acid (2) at about 31.823 min; Salicylic acid (3) at about 37.525 min; Trans-cinnamic acid (4) at about 40.006 min; Quercitrin (5) at about 33.052 min; Quercetin (6) at about 39,300 min; Chlorogenic acid (7) at about 26.078 min; Isoquercitrin (8) at about 30.762 min; At about 30.974 min, a peak corresponding to the hyper-
또한, 어성초 에탄올 추출물은 보유시간 약 33.052분에서 퀘르시트린(5); 약 39.321분에서 쿼세틴(6); 약 26.119분에서 클로로겐산(7); 약 30.802분에서 이소퀘르시트린(8); 약 31.037분에서 하이퍼로사이드(9)에 해당하는 피크가 관찰되었다(도 2D~F). 어성초 에탄올 추출물 및 어성초 에탄올 추출물의 분획물의 HPLC 결과에 따른 각 성분의 정량 평가 결과는 하기 표 2에 나타내었다. In addition, the extracts of Hwasungcho extracts showed quercitrin (5) at a retention time of about 33.052 min; Quercetin (6) at about 39.321 min; Chlorogenic acid (7) at about 26.119 min; Isoquercitrin (8) at about 30.802 min; At about 31.037 min, a peak corresponding to the
실시예 3. 어성초 추출물의 세포독성 분석Example 3: Cytotoxicity analysis of Hwasungcho extract
3-1. 섬유아세포 독성 분석3-1. Fibroblast toxicity analysis
섬유아세포(fibroblast)는 콜라겐과 엘라스틴 등을 생산하고, 피부 조직의 진피를 구성한다. 콜라겐은 두피를 건강하게 하고 탈모 예방 및 모발을 건강하게 하는 기능을 하지만, 노화가 진행될수록 콜라겐, 엘라스틴 또는 섬유아세포 등의 구조 단백질량이 감소함에 따라 조직이 느슨해져서 탈모가 발생하기도 한다.Fibroblasts produce collagen and elastin, and constitute the dermis of the skin tissue. Although collagen acts to make scalp healthy and prevent hair loss and to make hair healthy, the amount of structural proteins such as collagen, elastin or fibroblast decreases as aging progresses, resulting in tissue loosening and hair loss.
섬유아세포에 어성초 에탄올 추출물을 10, 50, 100 및 500 ㎍/mL의 농도로 처리하여 세포 독성을 확인한 결과, 어성초 에탄올 추출물 10, 50, 100 및 500 ㎍/mL 농도에서 세포 생존율이 각각 100%, 107.3%, 109.6%, 128.2%인 것으로 확인되어 세포 독성이 없는 것을 확인하였다(도 3). 이를 통해, 어성초 에탄올 추출물이 섬유아세포의 증식에 영향을 줄 수 있으므로, 발모 및 탈모 제품 개발에 유용한 소재로 사용될 수 있을 것으로 사료되었다.
Cell viability was determined at the concentrations of 10, 50, 100, and 500 ㎍ / mL of Hwasungcho extracts by 100%, 100%, and 500 ㎍ / mL, respectively. 107.3%, 109.6%, and 128.2%, respectively, and it was confirmed that there was no cytotoxicity (FIG. 3). This suggests that ethanol extract of Hwasungcho may affect the proliferation of fibroblasts and thus may be useful as a useful material for the development of hair growth and hair loss products.
3-2. 모유두세포 독성 분석3-2. Toothpick cytotoxicity analysis
인간 모유두세포(Human follicle dermal papilla cell, HFDPC)는 모발이 성장할 수 있는 신호를 모모세포(hair matrix cell)에 전달하여 모발을 형성하게 함으로써 발모와 탈모에 크게 관여한다.Human follicle dermal papilla cells (HFDPCs) are involved in hair growth and hair loss by allowing hair growth signals to be transferred to hair matrix cells to form hair.
인간 모유두세포에 어성초 에탄올 추출물을 10, 50, 100 및 500 ㎍/mL의 농도로 처리하여 24시간 후에 세포 독성을 확인한 결과, 어성초 에탄올 추출물 10, 50, 100 및 500 ㎍/mL 농도에서 세포 생존율이 각각 101.1%, 99.6%, 98.4%, 96.6%인 것으로 확인되어, 세포 독성이 없는 것을 확인하였다(도 4).
Cell viability of human dermal papilla cells at 10, 50, 100, and 500 ㎍ / mL concentrations of 10, 50, 100, and 500 ㎍ / Were found to be 101.1%, 99.6%, 98.4%, and 96.6%, respectively, indicating no cytotoxicity (Fig. 4).
3-3. 지방전구세포(3T3-L1) 독성 분석3-3. Toxicity analysis of lipogenic precursor cells (3T3-L1)
지방전구세포(3T3-L1)에 어성초 에탄올 추출물을 10, 50, 100 및 500 ㎍/mL의 농도로 처리하여 24시간 후에 세포 독성을 확인한 결과, 어성초 에탄올 추출물 10, 50 및 100 ㎍/mL 농도에서 세포 생존율이 각각 97.3%, 98.4%, 95.6%로 확인되었으나, 500 ㎍/mL 농도에서는 세포 생존율은 74.5%로 확인되어, 지방전구세포에 서는 어성초 에탄올 추출물이 100 ㎍/mL까지는 독성이 없는 것으로 확인되었다(도 5).
The cytotoxicity was examined 24 hours after treatment of liposome progenitor cells (3T3-L1) with 10, 50, 100 and 500 ㎍ / mL, The cell viability was 97.3%, 98.4% and 95.6%, respectively. However, the cell survival rate was found to be 74.5% at the concentration of 500 ㎍ / mL, and it was confirmed that the progesterone ethanol extract was not toxic up to 100 ㎍ / (Fig. 5).
실시예 4. 지방전구세포의 분화 현상 분석Example 4. Analysis of differentiation of lipid precursor cells
지방전구세포에 어성초 에탄올 추출물을 각각 10, 50, 100 및 500 ㎍/mL의 농도로 처리하여, 지방전구세포에서 지방세포로의 분화율을 측정하였다.The lipid precursor cells were treated with 10, 50, 100, and 500 ㎍ / mL, respectively, and the differentiation rate from adipose precursor cells to adipocytes was measured.
그 결과, 어성초 에탄올 추출물의 농도가 증가함에 따라 지방전구세포의 분화율도 증가하였으며, 특히 500 ㎍/mL 농도의 어성초 에탄올 추출물을 처리한 군은 분화유도 물질(DMⅡ; 1 μM Dexamethasone, 0.5 mM IBMX(IsoButhylMethylXanthine), 10 μg/mL Insulin)을 처리한 대조구에 비해 약 1.3배 증가하여 가장 높은 분화율을 나타내었다(도 6A). As a result, the differentiation rate of adipocyte progenitor cells increased as the concentration of the ethanol extract of the perennial herb was increased. Especially, the group treated with the ethanol extract of the perennial herb of 500 ㎍ / mL showed a differentiation inducer (DMII; 1 μM Dexamethasone, 0.5 mM IBMX IsoButhylMethylXanthine), 10 μg / mL Insulin) compared with the control, which was about 1.3 times higher than that of the control (FIG. 6A).
또한, 지방세포로의 분화를 형태학적으로 확인하기 위해 Oil-red O 염색을 실시한 결과, 500 ㎍/mL 농도의 어성초 에탄올 추출물을 처리한 군은 대조구에 비해 염색된 부분이 현저히 증가된 것으로 관찰되어, 지방전구세포가 지방세포로 분화가 많이 이루어진 것을 알 수 있었다(도 6B).In addition, the oil-red O staining was performed to confirm the morphological differentiation into adipocytes. As a result, it was observed that the 500 ㎍ / mL concentration of the ethanol extract of Hwasungcho significantly increased the stained portion compared with the control, And the lipid precursor cells were differentiated into adipocytes (FIG. 6B).
이를 통해, 어성초 에탄올 추출물이 지방전구세포(preadipocyte)에서 지방세포(adipocyte)로의 분화를 촉진시키는 것을 확인하였으며, 지방조직으로 구성된 피하조직(hypodermis)에서 체온조절, 완충 기능 및 모발에 영양분을 제공할 수 있을 것으로 사료되었다.
It was confirmed that the ethanol extract of Hwasungcho extract promoted the differentiation from preadipocyte to adipocyte, and the hypodermis composed of adipose tissue regulates body temperature, cushioning function, and provides nutrients to the hair .
실시예 5. 어성초 에탄올 추출물의 총 페놀 함량 분석Example 5 Analysis of Total Phenol Content of Ethanol Extract of Rhizosphere
폴리페놀성 화합물은 식물계에 널리 분포되어 있는 2차 대사산물 중 하나로 항산화활성, 항암 및 항균작용을 하는 생리활성 물질로 알려져 있으며, 대부분의 식물체들에서 항산화 활성을 나타내는 물질이다. The polyphenolic compound is one of the secondary metabolites widely distributed in the vegetable field. It is known as a physiologically active substance having antioxidative, anticancer and antimicrobial activities, and exhibits antioxidative activity in most plants.
어성초 에탄올 추출물의 동결건조물 내 총 폴리페놀(total pholyphenols) 함량을 분석한 결과, 총 폴리페놀 함량은 약 5.89%로 확인되었다.
The total polyphenols content in the lyophilized product of Hwasungcho extract was found to be 5.89%.
실시예 6. 어성초 에탄올 추출물의 항산화 활성 분석Example 6 Antioxidant Activity Analysis of Ethanol Extract of Hwasungcho
6-1. 전자공여능(Electron donating ability, EDA) 및 ABTS 라디칼 소거능(ABTS scavenging activity) 분석 6-1. Analysis of electron donating ability (EDA) and ABTS scavenging activity
어성초 에탄올 추출물의 동결건조물의 농도별(1, 5, 10, 50, 100 및 500 ㎍/mL) 전자공여능 및 ABTS 라디칼 소거능을 측정한 결과, 농도의존적으로 전자공여능(도 7A) 및 ABTS 라디칼 소거능(도 7B)이 증가하였으며, 특히 전자공여능의 경우 어성초 에탄올 추출물 50 ㎍/mL 이상의 농도에서는 비타민 C를 처리한 대조구와 비슷한 수준의 매우 강한 전자공여능을 나타내는 것으로 확인되었다.
The electron donating ability and the ABTS radical scavenging activity of each of the extracts of Hwasungcho extracts (1, 5, 10, 50, 100 and 500 ㎍ / mL) were measured in terms of the concentration of lyophilized product. The electron donating ability (FIG. 7A) and the ABTS radical scavenging ability 7B). Especially, in the case of electron donating ability, it was confirmed that the concentration of ethanol extract of Hwasungcho extract showed a strong electron donating ability similar to that of the control treated with vitamin C at a concentration of 50 ㎍ / mL or more.
6-2. 수퍼옥사이드 음이온 소거능(Superoxide anion radical scavenging activity, SOSA) 분석6-2. Superoxide anion radical scavenging activity (SOSA) analysis
어성초 에탄올 추출물의 동결건조물의 농도별(1, 5, 10, 50, 100 및 500 ㎍/mL)로 잔틴산화효소(xanthine oxidase)에 의해 생성되는 수퍼옥사이드 음이온 라디칼의 생성저해활성을 측정한 결과, 농도의존적으로 라디칼 생성저해활성이 증가하였으며, 특히 어성초 에탄올 추출물 10 ㎍/mL 이상의 농도에서는 비타민 C를 처리한 대조구보다 높은 라디칼 소거능을 나타내어 어성초 에탄올 추출물의 항산화 활성이 우수한 것을 확인하였다(도 7C).As a result of measuring the inhibitory activity of superoxide anion radicals produced by xanthine oxidase by concentration (1, 5, 10, 50, 100 and 500 ㎍ / mL) of lyophilized ethanol extract, (Fig. 7C). In particular, the ethanol extract of Hwasungcho showed higher radical scavenging activity than the control of vitamin C at a concentration of 10 ㎍ / mL or higher.
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