KR102512711B1 - Compositions comprising Ecklonia cava extract for acceleration of gastric motility - Google Patents
Compositions comprising Ecklonia cava extract for acceleration of gastric motility Download PDFInfo
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- KR102512711B1 KR102512711B1 KR1020200162758A KR20200162758A KR102512711B1 KR 102512711 B1 KR102512711 B1 KR 102512711B1 KR 1020200162758 A KR1020200162758 A KR 1020200162758A KR 20200162758 A KR20200162758 A KR 20200162758A KR 102512711 B1 KR102512711 B1 KR 102512711B1
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- South Korea
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- ecklonia cava
- present
- cava extract
- food
- extract
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- 229940018973 ecklonia cava extract Drugs 0.000 title claims abstract description 61
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Abstract
본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동 촉진용 식품 조성물과 사료 첨가용 조성물에 관한 것이다. 본 발명의 감태 추출물은 위장관 운동이 저하된 동물에 투여하였을 때 위장관 운동을 촉진하였고 이와 관련된 호르몬의 분비를 촉진하였으며 뮤신의 분비를 촉진시키는 효과를 보였을 뿐만 아니라, 천연물 유래 물질이기 때문에 부작용 없이 위장관 운동 저하 개선을 위한 장기 복용한 조성물로 활용될 수 있다.The present invention relates to a food composition for promoting gastrointestinal motility and a composition for adding feed containing Ecklonia cava extract as an active ingredient. When administered to animals with reduced gastrointestinal motility, the Ecklonia cava extract of the present invention promotes gastrointestinal motility, promotes the secretion of hormones related to it, and exhibits the effect of promoting mucin secretion. It can be used as a composition for long-term administration to improve degradation.
Description
본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동 촉진용 조성물에 관한 것이다.The present invention relates to a composition for promoting gastrointestinal motility comprising an extract of Ecklonia cava as an active ingredient.
위장관 운동 장애는 위식도 역류, 기능성 소화불량, 과민성 대장염, 위장관 마비, 구토, 당뇨병성 위장운동장애, 화학요법으로 인한 위장운동장애, 소화관 운동장애로 인한 장폐색, 수술 후 장폐색을 포함하는 수술 후 위장관 기능 장애, 근긴장성 이영양증으로 인한 위장관 운동 장애 등과 같이 정상적인 위장관 운동이 장애, 저해, 손상되어 있는 상태를 말한다.Gastrointestinal motility disorders include gastroesophageal reflux, functional dyspepsia, irritable colitis, gastrointestinal paralysis, vomiting, diabetic gastrointestinal motility disorders, chemotherapy-induced gastrointestinal motility disorders, ileus due to gastrointestinal motility disorders, and postoperative gastrointestinal tract obstruction, including postoperative ileus. It refers to a condition in which normal gastrointestinal motility is impaired, inhibited, or damaged, such as functional disorder or gastrointestinal motility disorder due to myotonic dystrophy.
위장관 운동 장애는 선진국은 물론 우리나라에서도 상당히 많이 발생하며, 다양한 증상을 보이는데, 그 중 특히 위장관 마비 증상이 주로 나타나 일상생활에 있어 고통과 불편함을 야기한다. 이러한 위장관 운동 장애는 위장관의 조직 병리학적 및 생화학적인 병변이 아닌 위장관 연동운동에 관련된 기능적 증상이다.Gastrointestinal motility disorder occurs quite frequently in Korea as well as in developed countries, and shows various symptoms, especially gastrointestinal paralysis symptoms, which cause pain and discomfort in daily life. Gastrointestinal motility disorders are functional symptoms related to gastrointestinal peristalsis rather than histopathological or biochemical lesions of the gastrointestinal tract.
현재 위장관 운동 장애는 뚜렷한 기질적 병변이 없는, 여러 가지 다양한 증상의 진단을 통해 진단하기 때문에 치료 또한 단순하지 않고, 대부분의 증상이 호전과 악화를 반복하며 음식, 스트레스 등에 의해 변화가 심하다. Currently, gastrointestinal motility disorder is diagnosed through diagnosis of various symptoms without clear organic lesions, so treatment is not simple, and most symptoms repeat improvement and deterioration, and changes are severe due to food and stress.
현재 대표적인 위장관 운동 촉진제로서, 돔페리돈 (Domperidone), 메토클로프라마이드(metoclopramide), 레보설프라이드(levosulpiride), 모사프라이드(mosapride), 이토프라이드(itopride) 등의 약물이 있다. Currently, representative gastrointestinal motility promoters include drugs such as domperidone, metoclopramide, levosulpiride, mosapride, and itopride.
그 중 메토클로프라미드(metoclopramide), 돔페리돈(domperidone), 시사프라이드(cisapride), 레보설프라이드(levosulpride), 에리스로마이신(erythromycin) 등의 위장운동 촉진제는 효과가 우수하다는 것이 여러 연구에서 알려져 있으며, 메토클로프라미드는 가장 흔히 사용되는 항도파민 약제로서, 중추신경계과 위장관의 도파민 수용체를 억제하는데 도파민은 위장관 운동을 억제하는 역할을 한다. 메토클로프라미드는 위십이지장의 운동을 증강시키고 위 배출기능을 호전시키며 진통작용이 있어 기능성 소화불량증과 같이 위장관 운동 장애에 따른 질환을 겪는 환자들의 증상을 개선하나, 장기간 사용시 증상의 호전효과가 감소하고 중추신경계의 도파민 수용체에 작용하여 많게는 약 24%의 환자에서 신경계통의 부작용을 일으킬 수 있다.Among them, it is known in several studies that gastrointestinal motility promoters such as metoclopramide, domperidone, cisapride, levosulpride, and erythromycin have excellent effects. , Metoclopramide is the most commonly used antidopamine drug, and it inhibits dopamine receptors in the central nervous system and gastrointestinal tract, and dopamine plays a role in inhibiting gastrointestinal motility. Metoclopramide enhances gastroduodenal motility, improves gastric emptying, and has analgesic action, improving symptoms of patients suffering from diseases caused by gastrointestinal motility disorders such as functional dyspepsia, but the improvement of symptoms decreases with long-term use. It acts on dopamine receptors in the central nervous system and can cause side effects of the nervous system in as many as about 24% of patients.
특히 위장관 운동 장애는 호전과 악화를 반복되기 때문에 장기복용을 필요로 하는 경우가 많은 바, 장기복용 시에도 인체에 안전한, 천연물 유래 위장관 운동 촉진제의 개발이 절실히 요구되고 있다.In particular, since gastrointestinal motility disorder repeats improvement and deterioration, long-term use is often required, and the development of a natural product-derived gastrointestinal motility promoter that is safe for the human body even during long-term use is urgently required.
본 발명자들은 천연물 유래 추출물이 저하된 위장관 운동을 촉진하는 것을 확인하여 본 발명을 완성하였다.The present inventors completed the present invention by confirming that the natural product-derived extract promotes reduced gastrointestinal motility.
따라서 본 발명의 목적은 위장관 운동(gastrointestinal motility) 촉진용 식품 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to provide a food composition for promoting gastrointestinal motility.
본 발명의 또 다른 목적은 위장관 운동 촉진용 식품 조성물의 제조 방법을 제공하는 것이다.Another object of the present invention is to provide a method for preparing a food composition for promoting gastrointestinal motility.
본 발명의 또 다른 목적은 위장관 운동 촉진용 사료 첨가용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for feed additives for promoting gastrointestinal motility.
상기 목적을 달성하기 위하여, 본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동(gastrointestinal motility) 촉진용 식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a food composition for promoting gastrointestinal motility comprising an Ecklonia cava extract as an active ingredient.
또한 상기 또 다른 목적을 달성하기 위하여, 본 발명은 감태를 분쇄하여 감태 분말을 제조하는 감태 분쇄 단계; 상기 감태 분말을 순환 추출하는 추출 단계; 상기 추출된 감태 추출물을 여과하는 단계; 상기 여과된 감태 추출물을 건조 농축하는 단계; 및 상기 건조 농축된 감태 추출물을 용매에 희석하는 단계;를 포함하는 위장관 운동 촉진용 식품 조성물의 제조 방법을 제공한다.In addition, in order to achieve the above another object, the present invention comprises the steps of grinding Ecklonia cava to prepare Ecklonia cava powder; Extraction step of circulating extraction of the Ecklonia cava powder; Filtering the extracted Ecklonia cava extract; Drying and concentrating the filtered Ecklonia cava extract; and diluting the dried and concentrated Ecklonia cava extract in a solvent.
또한 상기 또 다른 목적을 달성하기 위하여, 본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동 촉진용 사료 첨가용 조성물을 제공한다.In addition, in order to achieve the above another object, the present invention provides a composition for adding feed to promote gastrointestinal motility, containing Ecklonia cava extract as an active ingredient.
본 발명에 따른 감태 추출물은 위장관 운동이 저하된 동물에 투여하였을 때 위장관 운동을 촉진하였고 이와 관련된 호르몬의 분비를 촉진하였으며 뮤신의 분비를 촉진시키는 효과를 보였으므로, 이를 위장관 운동 촉진용 식품 조성물 및 사료 첨가용 조성물 등으로 유용하게 활용할 수 있다.When administered to animals with reduced gastrointestinal motility, the Ecklonia cava extract according to the present invention promotes gastrointestinal motility, promotes the secretion of hormones related thereto, and exhibits the effect of stimulating mucin secretion. It can be usefully utilized as a composition for additives and the like.
도 1은 본 발명에 따른 감태 추출물의 구성 성분을 GC-MS(gas chromatograph-mass spectrometer)를 이용하여 확인한 결과이다.
도 2는 본 발명에 따른 감태 추출물의 구성 성분을 FTIR(Fourier transform infrared spectroscopy)을 이용하여 확인한 결과이다.
도 3은 본 발명에 따른 감태 추출물의 장운동 촉진 효과를 확인한 결과이다.
도 4는 본 발명에 따른 감태 추출물의 장 운동성 호르몬 분비 촉진 효과를 확인한 결과이다.
도 5는 본 발명에 따른 감태 추출물의 뮤신 분비 촉진 효과를 확인한 결과이다.
도 6은 본 발명에 따른 감태 추출물의 뮤신 관련 유전자 발현 촉진 효과를 확인한 결과이다.1 is a result of confirming the components of the Ecklonia cava extract according to the present invention using a gas chromatograph-mass spectrometer (GC-MS).
2 is a result of confirming the components of the Ecklonia cava extract according to the present invention using FTIR (Fourier transform infrared spectroscopy).
Figure 3 is the result of confirming the intestinal motility promoting effect of the Ecklonia cava extract according to the present invention.
Figure 4 is the result of confirming the intestinal motility hormone secretion promoting effect of Ecklonia cava extract according to the present invention.
5 is a result confirming the mucin secretion promoting effect of the Ecklonia cava extract according to the present invention.
Figure 6 is the result of confirming the mucin-related gene expression promoting effect of the Ecklonia cava extract according to the present invention.
이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동(gastrointestinal motility) 촉진용 식품 조성물을 제공한다.The present invention provides a food composition for promoting gastrointestinal motility comprising an Ecklonia cava extract as an active ingredient.
본 발명에 있어서 상기 감태는 학명 Ecklonia cava의 해조류로, 갈조식물 다시마목 미역과의 해조에 속하고, 줄기는 원주상이며 충분히 자란 것은 1m 이상 되는 것도 있다. 중앙부가 굵고 어릴 때는 속이 차 있으나 다 자란 뒤에는 중앙부가 비어 있기도 한다. 줄기의 상부는 차차 편평하게 되고 양측에서 우상엽이 나고 여기서 다시 호생한 우상의 소엽편을 낸다. 엽면에는 주름이 없다. 색은 갈색인데 건조하면 흑색으로 된다. 주로 제주도 일대 및 일부 남해안에 분포하며, 점심대의 수심 10m 내외의 깊은 곳에서 서식한다. 2-3년간 생장하는 다년생 식물로서, 전복의 주 먹이이기도 한 감태는 알긴산, 요오드 및 칼륨 등의 영양소가 많이 함유되어 있어 건강식품자원으로 주목받고 있다.In the present invention, the Ecklonia cava is a seaweed with the scientific name Ecklonia cava, and belongs to the seaweed of the brown algae plant kelp order seaweed family, and the stem is columnar and has a sufficiently grown one that is 1m or more. The central part is thick and full when young, but the central part is empty after adulthood. The upper part of the stem gradually becomes flat, and pinnate lobe comes out on both sides, and pinnate lobe grows again here. There are no wrinkles on the leaf surface. The color is brown, but turns black when dried. It is mainly distributed in the whole area of Jeju Island and some parts of the southern coast, and lives in the depths of around 10m in the lunch zone. As a perennial plant that grows for 2-3 years, Ecklonia cava, which is also the main food for abalone, is attracting attention as a health food resource because it contains a lot of nutrients such as alginate, iodine and potassium.
본 발명의 감태 추출물은 공지의 천연물 추출방법에 의하여 추출될 수 있는 것으로서, 추출시 사용되는 추출 용매의 종류는 당업계에 해조류 성분 추출 용매로서 알려진 것이라면 그 종류가 특별히 제한되지 않으나, 바람직하게는 물, 탄소수 1 내지 6의 저급 알코올 (예. 메탄올, 에탄올, 프로판올, 부탄올, 펜탄올, 헥산올, 글리세린, 프로필렌글리콜 및 1,3-부틸렌글리콜 등), 탄소수 1 내지 4개의 저급케톤 (예. 아세톤, 메틸에틸케톤 등), 탄소수 1 내지 4개의 저급 에스테르 (예. 에틸 아세테이트 및 부틸 아세테이트 등), 아임계 유체 및 초임계 유체로 이루어진 군에서 선택된 하나 이상의 용매에 의해 추출되는 것일 수 있으며, 보다 바람직하게는 물, 탄소수 1 내지 6의 저급 알코올, 아세톤, 에틸아세테이트, 아임계 유체 및 초임계 유체로 이루어진 군에서 선택된 하나 이상의 용매에 의해 추출되는 것일 수 있다. 가장 바람직하게는 물, 탄소수 1 내지 4의 저급 알코올 및 이들의 혼합 용매로 이루어지는 군에서 선택된 1 종 이상의 용매에 의해 추출되는 것일 수 있다.The Ecklonia cava extract of the present invention can be extracted by a known natural product extraction method, and the type of extraction solvent used during extraction is not particularly limited as long as it is known as a seaweed component extraction solvent in the art, but preferably water , lower alcohols having 1 to 6 carbon atoms (eg methanol, ethanol, propanol, butanol, pentanol, hexanol, glycerin, propylene glycol and 1,3-butylene glycol, etc.), lower ketones having 1 to 4 carbon atoms (eg. acetone, methyl ethyl ketone, etc.), lower esters having 1 to 4 carbon atoms (eg, ethyl acetate and butyl acetate, etc.), subcritical fluids, and supercritical fluids. Preferably, it may be extracted with at least one solvent selected from the group consisting of water, lower alcohol having 1 to 6 carbon atoms, acetone, ethyl acetate, subcritical fluid and supercritical fluid. Most preferably, it may be extracted with one or more solvents selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
상기 감태 추출물의 제조는, 추출 효율을 증대시키기 위해 전처리 과정을 포함할 수 있으며, 예를 들어 감태를 건조한 후 분쇄기로 분쇄하여 사용할 수 있다.Preparation of the Ecklonia cava extract may include a pretreatment process to increase the extraction efficiency. For example, Ecklonia cava may be dried and then pulverized with a grinder.
본 발명의 추출물의 추출 온도는 특별히 제한되지 아니하며, 예를 들어 0℃ 내지 150℃일 수 있으며, 바람직하게는 10℃ 내지 80℃일 수 있고, 보다 바람직하게는 30℃ 내지 70℃일 수 있다. 본 발명의 추출물 추출 시간은 추출 온도에 따라 당업자가 적절히 설정 및 변경 가능한 것으로서 특별히 제한되지 아니하며, 예를 들어 10분 내지 10일일 수 있으며, 바람직하게는 1시간 내지 72시간 일 수 있으며, 더욱 바람직하게는 시간 12 내지 24시간 일 수 있다.The extraction temperature of the extract of the present invention is not particularly limited, and may be, for example, 0 °C to 150 °C, preferably 10 °C to 80 °C, and more preferably 30 °C to 70 °C. The extraction time of the extract of the present invention is not particularly limited as it can be appropriately set and changed by those skilled in the art according to the extraction temperature, and may be, for example, 10 minutes to 10 days, preferably 1 hour to 72 hours, and more preferably may be between 12 and 24 hours.
본 발명 추출물은 공지의 천연물 추출법으로 추출될 수 있다. 예를 들어, 순환추출, 냉침추출, 상온추출, 열수추출, 환류냉각 추출, 가열 추출, 고압추출, 초음파 추출, 아임계 추출법, 초임계 추출법 등으로 추출할 수 있으며, 1회 내지 10회, 바람직하게는 2회 내지 7회 반복 추출할 수 있다.The extract of the present invention may be extracted by a known natural product extraction method. For example, it can be extracted by circulation extraction, cold extraction, room temperature extraction, hot water extraction, reflux cooling extraction, heating extraction, high pressure extraction, ultrasonic extraction, subcritical extraction, supercritical extraction, etc., preferably 1 to 10 times. Preferably, extraction may be repeated 2 to 7 times.
또한 본 발명의 감태 추출물의 제조시 처리, 보관 등의 용이함을 위하여 여과 과정, 농축 및 정제과정, 건조과정, 동결과정, 분말화 과정 등이 임의로 추가될 수 있다.In addition, filtration process, concentration and purification process, drying process, freezing process, pulverization process, etc. may be optionally added for ease of processing and storage during the preparation of the Ecklonia cava extract of the present invention.
상기 여과 과정은 공지의 여과 방법에 의할 수 있으며 이에 제한되지 않으나, 예를 들어 여과망 또는 마이크로필터를 이용한 여과, 원심분리 및 분액깔때기를 이용할 수 있다. 상기 농축 과정은 공지의 농축 방법에 의할 수 있으며 이에 제한되지는 않으나, 예를 들어 침전농축, 증발농축, 감압농축, 한외여과법, 역삼투법 및 원심분리법을 이용하여 농축할 수 있다.The filtration process may be performed by a known filtration method, but is not limited thereto, and for example, filtration using a filter net or microfilter, centrifugation, and separatory funnel may be used. The concentration process may be by a known concentration method, but is not limited thereto, and may be concentrated using, for example, precipitation concentration, evaporation concentration, vacuum concentration, ultrafiltration, reverse osmosis, and centrifugation.
상기 건조 과정은 공지의 건조 방법에 의할 수 있으며 이에 제한되지 아니하나, 예를 들어 동결 건조, 분무 건조 또는 열풍건조 일 수 있다. 또한 분말화 과정은 공지의 분말화 방법에 의할 수 있으며 이에 제한되지 아니하나, 예를 들어 덱스트린 포접 등의 과정에 의해 분말화 될 수 있다.The drying process may be by a known drying method, but is not limited thereto, and may be, for example, freeze drying, spray drying or hot air drying. In addition, the powdering process may be by a known powdering method, but is not limited thereto, and may be powdered by, for example, dextrin inclusion or the like.
본 발명의 감태 추출물은 위장관 운동 촉진은 위장관 연동 운동(gastric motility) 작용을 촉진하는 것을 특징으로 한다.The Ecklonia cava extract of the present invention is characterized by promoting gastrointestinal motility.
본 발명의 일 실시예에 따르면, 본 발명에 따른 감태 추출물을 위장관 운동 능력이 저해된 동물 모델에 처리하였을 때 장내 음식물 이동률(Gastrointestinal transit)이 농도 의존적으로 증가한 것을 확인하였다. According to one embodiment of the present invention, it was confirmed that gastrointestinal transit increased in a concentration-dependent manner when the Ecklonia cava extract according to the present invention was treated in an animal model with impaired gastrointestinal motility.
본 발명의 “식품”은, 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food) 및 식품 첨가제(food additives) 등의 모든 형태로 제조할 수 있다. 예를 들면, 건강식품으로는 본 발명의 감태 추출물을 차, 쥬스 및 드링크의 형태로 제조하여 음용하도록 하거나, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 기능성 식품으로는 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마말레이드 등), 어류, 육류 및 그 가공식품(예: 햄, 소시지, 콘비프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치츠 등), 식용식물유지, 마가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 본 발명의 감태 추출물을 첨가하여 제조할 수 있다.The "food" of the present invention can be prepared in any form, such as functional food, nutritional supplement, health food, and food additives. For example, as a health food, the Ecklonia cava extract of the present invention can be prepared in the form of tea, juice and drink to be consumed, or granulated, encapsulated and powdered to be consumed. In addition, functional foods include beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruits, canned fruits, jams, marmalades, etc.), fish, meat and their processed foods (e.g. ham, sausages, corned beef, etc.) , Breads and noodles (e.g. udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine, vegetable protein, retort food It can be prepared by adding the Ecklonia cava extract of the present invention to frozen foods, various seasonings (eg, soybean paste, soy sauce, sauce, etc.).
본 발명의 감태 추출물을 식품 조성물의 형태로 사용할 경우, 상기 식품 조성물을 그대로 사용하거나 또는 다른 식품에 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 이는 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.When the Ecklonia cava extract of the present invention is used in the form of a food composition, the food composition can be used as it is, added to other foods, or used together with other foods or food ingredients, which can be appropriately used according to conventional methods. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).
본 발명의 식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100㎖ 당 일반적으로 약 0.01 내지 0.4g, 바람직하게는 약 0.02 내지 0.03g이다.The food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional components. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. . The proportion of the natural carbohydrate is generally about 0.01 to 0.4 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 식품 조성물은 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, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 식품 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 본 발명의 조성물 100 중량부 당 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이나, 이에 제한되는 것은 아니다.In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , carbonation agents used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is generally selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention, but is not limited thereto.
또한 본 발명은 감태를 분쇄하여 감태 분말을 제조하는 감태 분쇄 단계; 상기 감태 분말을 순환 추출하는 추출 단계; 상기 추출된 감태 추출물을 여과하는 단계; 상기 여과된 감태 추출물을 건조 농축하는 단계; 및 상기 건조 농축된 감태 추출물을 용매에 희석하는 단계; 를 포함하는 위장관 운동 촉진용 식품 조성물의 제조 방법을 제공한다.In addition, the present invention comprises the steps of pulverizing Ecklonia cava to prepare Ecklonia cava powder; Extraction step of circulating extraction of the Ecklonia cava powder; Filtering the extracted Ecklonia cava extract; Drying and concentrating the filtered Ecklonia cava extract; and diluting the dried concentrated Ecklonia cava extract in a solvent; It provides a method for producing a food composition for promoting gastrointestinal motility comprising a.
상기 순환 추출은 30 내지 70℃에서 12 내지 48시간 동안 수행할 수 있으나, 이에 제한되는 것은 아니며 보다 바람직하게는 50℃에서 24시간 동안 수행할 수 있으나 이에 제한되는 것은 아니다.The circulation extraction may be performed at 30 to 70° C. for 12 to 48 hours, but is not limited thereto, and more preferably at 50° C. for 24 hours, but is not limited thereto.
더불어 상기 여과는 0.2 내지 0.6 ㎛의 여과 사이즈를 갖는 여과필터를 이용하여 수행할 수 있으나, 이에 제한되는 것은 아니며 보다 바람직하게는 0.4㎛의 여과 사이즈를 갖는 여과필터를 이용하여 수행할 수 있으나, 이에 제한되는 것은 아니다.In addition, the filtration may be performed using a filtration filter having a filtration size of 0.2 to 0.6 μm, but is not limited thereto, and more preferably may be performed using a filtration filter having a filtration size of 0.4 μm. It is not limited.
또한 본 발명은 감태 추출물을 유효성분으로 포함하는 위장관 운동 촉진용 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for adding feed to promote gastrointestinal motility, comprising an Ecklonia cava extract as an active ingredient.
본 발명의 용어 "사료"란, 동물이 먹는 임의의 천연 또는 인공 구정식, 한끼식 등 또는 상기 한끼식의 성분을 의미하며, 본 발명에 따른 위장관 운동 촉진용 조성물을 유효성분으로 포함하는 사료는 당업계에 공지된 다양한 형태의 사료로 제조가능하며, 바람직하게는 농후 사료, 조사료 및/또는 특수사료가 포함될 수 있으나, 이로 제한되는 것은 아니다.The term "feed" of the present invention refers to any natural or artificial oral diet, one meal, etc., or a component of the one meal meal eaten by animals, and the feed containing the composition for promoting gastrointestinal motility according to the present invention as an active ingredient It can be prepared with various types of feed known in the art, preferably, concentrated feed, roughage and / or special feed may be included, but is not limited thereto.
본 발명에서 용어, “사료첨가용 조성물”은 “사료첨가제”를 의미할 수 있으며, 이는 동물에 있어 질병 증상의 완화, 영양소 보충 및 체중감소 예방, 사료 내 섬유소의 소화 이용성 증진, 유질개선, 번식장애 예방 및 수태율 향상, 하절기 고온 스트레스 예방 등 다양한 효과를 목적으로 사료에 첨가하는 물질을 포함한다.In the present invention, the term "composition for feed additive" may mean "feed additive", which alleviates symptoms of disease in animals, supplements nutrients and prevents weight loss, enhances the digestibility of fiber in feed, improves milk quality, and breeds. It includes substances added to feed for the purpose of various effects, such as preventing disorders, improving conception rates, and preventing high-temperature stress in the summer.
본 발명의 사료첨가용 조성물은 사료관리법상의 보조사료에 해당하며, 탄산수소나트륨, 벤토나이트(bentonite), 산화마그네슘, 복합광물질 등의 광물질제제, 아연, 구리, 코발트, 셀레늄 등의 미량 광물질인 미네랄제제, 케로틴, 비타민 A D, E, 니코틴산, 비타민 B 복합체 등의 비타민제, 메티오닌, 라이신 등의 보호아미노산제, 지방산 칼슘염 등의 보호지방산제, 생균제(유산균제), 효모배양물, 곰팡이 발효물 등의 생균, 효모제 등이 추가로 포함될 수 있다.The composition for feed additive of the present invention corresponds to supplementary feed under the Feed Management Act, and is a mineral preparation such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, and a trace mineral preparation such as zinc, copper, cobalt, and selenium. , vitamins such as kerotene, vitamins A, D, E, nicotinic acid, and vitamin B complex, protected amino acids such as methionine and lysine, protected fatty acids such as calcium salts of fatty acids, probiotics (lactic acid bacteria), yeast cultures, and fungal fermentation products. Probiotics, yeasts, and the like may be further included.
상기 사료 중 농후사료로는 밀, 귀리, 옥수수 등의 곡류를 포함하는 종자열매류, 곡물을 정제하고 얻는 부산물로서 쌀겨, 밀기울, 보릿겨 등을 포함하는 겨류, 콩, 유체, 깨, 아마인, 코코야자 등을 채유하고 얻는 부산물인 깻묵류와 고구마, 감자 등에서 녹말을 뺀 나머지인 녹말찌꺼기의 주성분인 잔존녹말질류 등의 찌꺼기류, 어분, 물고기찌꺼기, 어류에서 얻은 신선한 액상물(液狀物)을 농축시킨 것인 피시솔루블(fish soluble), 육분(肉粉), 혈분, 우모분, 탈지분유, 우유에서 치즈, 탈지유에서 카제인을 제조할 때의 잔액인 훼이(whey)를 건조한 건조훼이 등의 동물질사료, 효모, 클로렐라, 해조류가 있으나 이에 제한되지 않는다.Among the feeds, the enriched feed includes seed fruits including grains such as wheat, oats and corn, bran including rice bran, bran, barley bran, etc. as a by-product obtained by refining grains, soybeans, fluids, sesame seeds, linseed, coco Sesame cakes, which are by-products obtained from oil extraction of palms, and residual starches, which are the main component of starch residues, which are the remainder after removing starch from sweet potatoes and potatoes, fish meal, fish residues, and fresh liquid obtained from fish Animal substances such as concentrated fish soluble, meat meal, blood meal, feather meal, skim milk powder, dried whey, which is the balance when manufacturing cheese from milk and casein from skim milk, dried Feed, yeast, chlorella, algae, but not limited thereto.
상기 사료 중 조사료로는 야초, 목초, 풋베기 등의 생초(生草)사료, 사료용 순무, 사료용 비트, 순무의 일종인 루터베어거 등 의 뿌리채소류, 생초, 풋베기작물, 곡실(穀實) 등을 사일로에 채워 놓고 젖산발효시킨 저장사료인 사일리지(silage), 야초, 목초를 베어 건조시킨 건초, 종축용(種畜用) 작물의 짚, 콩과 식물의 나뭇잎이 있으며, 이에 제한되지 않는다. 특수사료에는 굴껍데기, 암염 등의 미네랄 사료, 요소나 그 유도체인 디우레이드이소부탄 등의 요소사료, 천연사료원료만을 배합했을 때 부족하기 쉬운 성분을 보충하거나, 사료의 저장성을 높이기 위해서 배합사료에 미량으로 첨가하는 물질인 사료첨가물, 식이보조제가 있으나 이에 제한되지 않는다.Forages among the feed include raw grass feed such as wild grass, grass, and green cutting, turnip for feed, beet for feed, root vegetables such as Lutherbearer, a kind of turnip, raw grass, green crops, grain Silage, which is a stored feed made by lactic acid fermentation by filling the back into a silo, wild grass, hay dried by cutting grass, straw of breeding crops, and leaves of legumes, but is not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that are likely to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storability of feeds. There are feed additives and dietary supplements, which are substances added in small amounts, but are not limited thereto.
본 발명에 따른 상기 사료첨가용 조성물은 당업계에 공지된 다양한 사료 제조방법에 따라 적절한 유효 농도 범위에서 감태 추출물을 첨가하여 제조 가능하다.The feed additive composition according to the present invention can be prepared by adding Ecklonia cava extract in an appropriate effective concentration range according to various feed preparation methods known in the art.
본 발명에 따른 사료 첨가제는 위장관 운동 촉진을 목적으로 하는 개체라면 제한없이 적용가능하다. 예를 들면, 원숭이, 개, 고양이, 토끼, 모르모트, 랫트, 마우스, 소, 양, 돼지, 염소 등과 같은 비인간 동물, 조류 및 어류 등 어느 개체에도 적용이 가능하다.The feed additive according to the present invention can be applied without limitation to any individual for the purpose of promoting gastrointestinal motility. For example, it can be applied to any object such as non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds and fish.
상술한 본 발명의 내용은 상호 모순되지 않는 한, 서로 동일하게 적용되며, 당해 기술분야의 통상의 기술자가 적절한 변경을 가해 실시하는 것 또한 본 발명의 범주에 포함된다.The contents of the present invention described above are equally applied to each other unless contradictory to each other, and implementation by adding appropriate changes by a person skilled in the art is also included in the scope of the present invention.
이하 본 발명을 실시예를 통해 상세하게 설명하나 본 발명의 범위가 하기 실시예로만 한정되는 것은 아니다. Hereinafter, the present invention will be described in detail through examples, but the scope of the present invention is not limited only to the following examples.
실시예 1. 감태 추출물의 제조Example 1. Preparation of Ecklonia cava extract
감태의 건조 샘플은 Parajeju Com. (한국 제주)에서 구입하였고, 감태(WPC-19-001) 바우처 표본은 부산 대학교 웰빙 RIS 센터 기능성 소재 은행에 기탁하였다. 건조된 감태를 분쇄기(MF-3100S, Hanil Electric Co., 서울)를 이용하여 분쇄한 후, 50 ℃에서 24 시간동안 순환추출장비(SHWB-30/45, Woori Science Instrument Co., 포천)를 사용하여 고정액 비율(감태 건조 분말:dH2O, 1:10)로 추출하였다. 그 후 추출물을 0.4 μ필터에 통과시켜 여과한 후 회전 증발기(EYELA, 도쿄, 일본)를 이용하여 건조하고 사용할 때까지 -80 ℃에서 보관했다. 그리고 실험 농도에 맞추어 저농도는 200mg, 고농도는 400mg으로 dH2O에 희석하여 감태 추출물을 제조하였다.Dried samples of Ecklonia were obtained from Parajeju Com. (Jeju, Korea), and the voucher sample of Ecklonia cava (WPC-19-001) was deposited in the Functional Material Bank of the Well-Being RIS Center, Pusan National University. After pulverizing the dried Ecklonia cava using a grinder (MF-3100S, Hanil Electric Co., Seoul), use circulation extraction equipment (SHWB-30/45, Woori Science Instrument Co., Pocheon) for 24 hours at 50 ℃. and extracted with a fixed solution ratio (e.g. dry powder: dH 2 O, 1:10). Thereafter, the extract was filtered through a 0.4 μ filter, dried using a rotary evaporator (EYELA, Tokyo, Japan), and stored at -80 °C until use. In addition, according to the experimental concentration, the low concentration was 200 mg and the high concentration was 400 mg, diluted in dH 2 O to prepare Ecklonia cava extract.
실시예 2. 감태 추출물의 성분 분석Example 2. Component analysis of Ecklonia cava extract
상기 실시예 1에서 제조한 감태 추출물의 총 탄닌 함량을 바닐린(Vanillin) 방법을 이용하여 측정하였다. 먼저, 감태 추출물(100 uL)을 500 uL의 혼합 용액 (1% vanillin/MeOH과 8% HCl/MeOH, 1:1 비율)과 혼합한 다음 30℃에서 20분 동안 배양했다. 배양 후 VersaMax 플레이트 리더 (Molecular Devices)를 사용하여 500nm에서 흡광도를 측정하였다. 총 축합된 탄닌 함량은 정제된 (+)-카테킨 수화물 기준((+)-catechin hydrate standard (Sigma-Aldrich Co.))을 사용하여 구성된 검량선으로부터 계산하였고, 그 결과를 하기 표 1에 나타내었다. The total tannin content of the Ecklonia cava extract prepared in Example 1 was measured using the Vanillin method. First, Ecklonia cava extract (100 uL) was mixed with 500 uL of a mixed solution (1% vanillin/MeOH and 8% HCl/MeOH, 1:1 ratio) and then incubated at 30°C for 20 minutes. After culturing, absorbance was measured at 500 nm using a VersaMax plate reader (Molecular Devices). The total condensed tannin content was calculated from a calibration curve constructed using a purified (+)-catechin hydrate standard (Sigma-Aldrich Co.), and the results are shown in Table 1 below.
감태 추출물의 총 축합된 탄닌 함량은 326.5±12.2 mg/g으로, 총 플라보노이드 함량(34.9 mg/g)과 총 페놀 함량(74.6 mg/g)보다 10배가량 높은 것으로 확인되었다.The total condensed tannin content of Ecklonia cava extract was 326.5 ± 12.2 mg/g, which was 10 times higher than the total flavonoid content (34.9 mg/g) and total phenol content (74.6 mg/g).
또한, 감태 추출물의 구성 성분을 GC-MS(gas chromatograph-mass spectrometer) 및 FTIR(Fourier transform infrared spectroscopy)을 이용하여 분석하였다. 자동 열탈착장치(ATD 400, Perkin Elmer, UK)가 장착된 GC-MS (QP-2010A, Shimadzu, Japan)를 사용하였고, 모세관 컬럼 AT-1은 60 m ×0.32 mm ×1.0 mm이고 질량범위는 20 - 350 m/z이었다. FTIR 분광법(Impact 400D, Nicolet, Madison, WI, USA)은 각 IR 분광계 샘플에 대해 400 ~ 4000cm-1 사이의 4cm-1 해상도에서 32개의 스캔이 투과 모드에서 수집되었다. In addition, components of Ecklonia cava extract were analyzed using GC-MS (gas chromatograph-mass spectrometer) and FTIR (Fourier transform infrared spectroscopy). A GC-MS (QP-2010A, Shimadzu, Japan) equipped with an automatic thermal desorption device (ATD 400, Perkin Elmer, UK) was used, and the capillary column AT-1 was 60 m × 0.32 mm × 1.0 mm and had a mass range of 20 - It was 350 m/z. For FTIR spectroscopy (Impact 400D, Nicolet, Madison, WI, USA), 32 scans were collected in transmission mode at 4 cm −1 resolution between 400 and 4000 cm −1 for each IR spectrometer sample.
그 결과 도 1에 나타낸 바와 같이, 감태 추출물의 주요 구성 요소는 카르바민산(carbamic acid), 에탄올(ethanol), 아세톤(acetone), 톨루엔(toluene), 데칸알(decanal), 실리케이트 음이온 사합체(silicate anion tetramer), n-노나날(n-nonanal)로 확인되었다. 더불어 FTIR을 이용하여 확인한 결과 도 2와 같이, 알코올(3274cm-1), 셀룰로오스 및 탄닌 (2971 및 2948cm-1), 리그닌 및 탄닌 (1608 및 1521 cm-1) 등이 감태 추출물에 존재함을 확인하였다.As a result, as shown in FIG. 1, the main components of the Ecklonia cava extract are carbamic acid, ethanol, acetone, toluene, decanal, silicate anion tetramer ( silicate anion tetramer) and n-nonanal. In addition, as a result of confirmation using FTIR, as shown in FIG. 2, it was confirmed that alcohol (3274 cm -1 ), cellulose and tannin (2971 and 2948 cm -1 ), lignin and tannin (1608 and 1521 cm -1 ), etc. are present in the Ecklonia cava extract. did
실시예 3. 감태 추출물의 장운동 촉진 효과 확인Example 3. Confirmation of intestinal motility promoting effect of Ecklonia cava extract
감태 추출물의 장 운동성 촉진 기능을 확인하기 위하여 장내 음식물 이동률(Gastrointestinal transit)을 분석하였다. 먼저 SD 랫트(rat)를 세 그룹으로 분류하였다(대조군: Vehicle 그룹, 저농도 감태 추출물 투여군(WELo군), 고농도 감태 추출물 투여군(WEHi군). 그런 후 각 실험군의 랫트에 로페라미드(Loperamide)를 투여하여 위장관 운동을 저하시키고 목탄(charcoal(0.5% 수성 메틸셀룰로오스(aqueous methylcellulose)에 3%의 활성탄(activated charcoal) 현탁)(Sigma-Aldrich Co.))를 3ml씩 경구투여한 후, 30분 후에 안락사시켰다. 안락사된 랫트의 장을 채취하여 전체 장 길이와 활성탄의 이동거리를 측정하였고, 장내 음식물의 이동률은 하기와 같이 계산하였다.Gastrointestinal transit was analyzed to confirm the intestinal motility promoting function of Ecklonia cava extract. First, SD rats were classified into three groups (control group: vehicle group, low concentration Ecklonia cava extract treatment group (WELo group), and high concentration Ecklonia cava extract treatment group (WEHi group). Then, Loperamide was administered to rats in each experimental group. After administration to reduce gastrointestinal motility, charcoal (charcoal (activated charcoal suspension of 3% in 0.5% aqueous methylcellulose) (Sigma-Aldrich Co.)) was orally administered by 3ml each, 30 minutes later The intestines of the euthanized rats were collected to measure the total intestinal length and the movement distance of activated carbon, and the movement rate of food in the intestine was calculated as follows.
[수학식 1][Equation 1]
장내 음식물의 이동률(%) = [(총 소장(small intestine)의 길이 - 활성탄의 이동거리)/총 소장의 길이)] ×100 Movement rate of food in the intestine (%) = [(total small intestine length - activated charcoal travel distance)/total small intestine length)] × 100
그 결과 도 3과 같이, 위장관 운동성이 저해된 Vehicle 그룹에 비해 감태를 투여한 WELo군과 WEHi군에서는 농도 의존적으로 활성탄의 이동이 더 증가하였다. 또한 장의 길이도 감태 추출물을 처리한 동물에서 회복됨을 확인하였다. 따라서, 본 발명에 따른 감태 추출물이 저하된 장 운동성을 회복시킬 수 있음을 확인하였다.As a result, as shown in FIG. 3, the movement of activated carbon was increased in a concentration-dependent manner in the WELo group and the WEHi group administered with Ecklonia cava compared to the Vehicle group in which gastrointestinal motility was inhibited. In addition, it was confirmed that the intestinal length was restored in animals treated with Ecklonia cava extract. Therefore, it was confirmed that the Ecklonia cava extract according to the present invention could restore the decreased intestinal motility.
실시예 4. 감태 추출물의 장 운동성 호르몬 분비 촉진 효과 확인Example 4. Confirmation of intestinal motility hormone secretion stimulating effect of Ecklonia cava extract
대장에서 분비되는 장 운동성 호르몬의 발현에 있어서, 본 발명에 따른 감태 추출물의 효과를 확인하였다. 장 운동성 호르몬인 콜레시스토키닌(cholecystokinin, CCK), 가스트린(gastrin) 및 소마토스타틴(somatostatin, SS)의 발현을 측정하기 위하여 ELISA kit (Cusabio Biotech Co., Ltd., Wuhan, 중국)를 사용하였다. 대장 조직 100 mg을 정량하여 얼음에 보관한 인산완충용액(phosphate buffered saline, PBS, pH 7.2-7.4) 1㎖을 첨가하여 분쇄한 후 4℃에서 1,000 xg의 속도로 5분간 원심분리하고 상층액을 수득하였다. 이에 상기 3 종류의 호르몬 항체(각 웰에 별도로)를 첨가한 후, 37℃에서 1시간 배양한 후에 HRP-스트렙타비딘 용액(HRP-Streptavidin solution)을 첨가하고 37℃에서 1시간 동안 추가로 배양하였다. 그리고 TMP 원스텝 기질 시약(TMP One-Step Substrate Reagent)을 첨가하고 37℃에서 30분간 반응시킨 후에 정지(stop) 용액을 첨가하여 반응을 정지시켰다. 그 후 Molecular Devices VERSA max Plate reader (Sunnyvale, CA, 미국)를 이용하여 흡광도 450nm에서 측정하였다.The effect of the Ecklonia cava extract according to the present invention on the expression of intestinal motility hormone secreted from the large intestine was confirmed. An ELISA kit (Cusabio Biotech Co., Ltd., Wuhan, China) was used to measure the expression of cholecystokinin (CCK), gastrin, and somatostatin (SS), which are intestinal motility hormones. 100 mg of colon tissue was quantified, pulverized by adding 1 ml of phosphate buffered saline (PBS, pH 7.2-7.4) stored on ice, centrifuged at 4°C at 1,000 xg for 5 minutes, and the supernatant was removed. obtained. After adding the above three types of hormone antibodies (separately to each well), incubation at 37°C for 1 hour, HRP-Streptavidin solution was added, and further incubation was performed at 37°C for 1 hour. did Then, the reaction was stopped by adding a stop solution after adding TMP One-Step Substrate Reagent and reacting at 37° C. for 30 minutes. Then, absorbance was measured at 450 nm using a Molecular Devices VERSA max Plate reader (Sunnyvale, CA, USA).
그 결과 도 4와 같이, 위장관 운동이 저해된 Vehicle 그룹에서는 3 종류의 장내 호르몬 농도가 유의적으로 감소한 반면, 본 발명에 따른 감태 추출물을 투여한 WELo군과 WEHi군에서는 농도 의존적으로 증가하였다. 따라서, 본 발명에 따른 감태 추출물은 위장관 운동 감소로 인한, 저하된 장 운동성 호르몬의 농도를 회복시켜주는 효과가 있음을 확인하였다.As a result, as shown in FIG. 4 , concentrations of three kinds of intestinal hormones were significantly decreased in the Vehicle group in which gastrointestinal motility was inhibited, whereas concentration-dependent increases were observed in the WELo group and the WEHi group administered with the Ecklonia cava extract according to the present invention. Therefore, it was confirmed that the Ecklonia cava extract according to the present invention has the effect of restoring the reduced concentration of intestinal motility hormone due to the decrease in gastrointestinal motility.
실시예 5. 감태 추출물의 뮤신 분비 촉진 효과 확인Example 5. Confirmation of mucin secretion promoting effect of Ecklonia cava extract
대장의 뮤신 분비에 있어서, 본 발명의 감태 추출물이 미치는 영향을 확인하기 위하여 공지된 방법에 따라 알시안 청색 염색(alcian blue staining)을 실시하였다. 알시안 청색 염색 분석은 박절된 조직을 자일렌(xylene)에 3분씩 3번 담그고, 100% 알코올에 각각 2분, 1분 담근 후 95, 80, 70% 알코올에 각각 1분, 30초씩 담근 후에 물로 1분 세척한 뒤 알시안 청색 염색 키트(Alcian Blue Stain kit (IHC WORLD, Woodstock, MD, 미국))를 사용하여 염색하였다. 슬라이드를 고정시킨 후 건조하여 BX50F-3 현미경 (Olympus, Tokyo, 일본)을 이용하여 관찰하였으며, Leica Application Suite (Leica Microsystems, Wetzlar, 독일)를 이용하여 분석하였다.In order to confirm the effect of the Ecklonia cava extract of the present invention on mucin secretion in the large intestine, alcian blue staining was performed according to a known method. Alcian blue staining analysis was performed by dipping the cut tissue in xylene three times for 3 minutes each, then in 100% alcohol for 2 minutes and 1 minute, respectively, and then in 95, 80, and 70% alcohol for 1 minute and 30 seconds, respectively. After washing with water for 1 minute, it was stained using an Alcian Blue Stain kit (IHC WORLD, Woodstock, MD, USA). The slides were fixed, dried, observed using a BX50F-3 microscope (Olympus, Tokyo, Japan), and analyzed using the Leica Application Suite (Leica Microsystems, Wetzlar, Germany).
그 결과 도 5에 나타낸 바와 같이, 위장관 운동이 저해된 Vehicle 그룹에서 대장 장샘(crypt)의 뮤신 분비는 감소하였지만 본 발명에 따른 감태 추출물을 투여한 WELo군과 WEHi군에서는 유의적으로 증가하는 것을 확인하였다. 그러므로 본 발명에 따른 감태 추출물은 뮤신 분비를 촉진하는 효과를 가짐을 확인하였다.As a result, as shown in FIG. 5, mucin secretion of the colon crypt decreased in the Vehicle group in which gastrointestinal motility was inhibited, but significantly increased in the WELo group and the WEHi group administered with the Ecklonia cava extract according to the present invention. did Therefore, it was confirmed that the Ecklonia cava extract according to the present invention has an effect of promoting mucin secretion.
실시예 6. 감태 추출물의 뮤신 관련 유전자 발현 촉진 효과 확인Example 6. Confirmation of mucin-related gene expression promoting effect of Ecklonia cava extract
본 발명에 따른 감태 추출물이 뮤신 유전자의 발현에 미치는 영향을 qRT-PCR을 수행하여 확인하였다. qRT-PCR은 랫트의 대장조직으로부터 RNA를 분리하여 실시하였다. SD랫트의 대장조직 300㎎을 적출하여 균질기를 이용하여 분쇄한 후 RNA를 추출하였다. RNAzol 추출물에 포함된 단백질은 클로로포름(chloroform)을 이용하여 추출한 후 아이소프로판올(isopropanol)을 이용하여 RNA를 침전시켜 수확하였다. 분리된 전체 RNA는 260nm에서 흡광도를 측정하고, 5㎍을 사용하였다. 먼저 5㎍의 RNA에 Oligo dT (invitrogen, 18418-012)을 처리하고, 70℃에서 10분 동안 반응시켜 Oligo dT를 RNA에 결합시켰다. 여기에 5x 완충용액, 10mM dNTP, 0.1 M DTT, Superscript II (Invitrogen, 18064-014, 200 U/㎕)를 첨가하여 실온에서 10분 동안 방치한 후 42 ℃에서 50분 동안 반응시켰다. 반응이 종료된 후 RNaseH(Invitrogen, 18021-071)를 처리하여 37℃에서 20분 동안 반응시켜 RNA를 분해하였다. qPCR은 획득한 cDNA 템플릿(2μL)과 하기의 프라이머를 포함하는 2x Power SYBR Green (6μL; Toyobo Life Science, Osaka, Japan)으로 수행하였다. 94℃(15초), 70℃(60초) 40회(cycle)로 수행하였으며, MUC2에 대한 프라이머(5’TCCTG TGGCA TCCAT GAAAC-3’β-액틴(β-actin)에 대한 프라이머(5’CGCAG CTCAG TAACA GTCCG-3’를 사용하였다. 더불어 PCR 증폭의 지수 단계에서 PCR 산물이 형광 강도 역치를 초과하는 반응주기를 역치주기(threshold cycle, Ct)로 간주하였다. 일정한 형광 강도에서 Cts를 비교하여 대상 유전자의 발현을 β-액틴과 비교하여 정량화하였다.The effect of the Ecklonia cava extract according to the present invention on the expression of mucin genes was confirmed by performing qRT-PCR. qRT-PCR was performed by isolating RNA from rat colon tissue. 300 mg of SD rat colon tissue was extracted, ground using a homogenizer, and then RNA was extracted. Proteins included in the RNAzol extract were extracted using chloroform and harvested by precipitating RNA using isopropanol. The isolated total RNA was measured for absorbance at 260 nm, and 5 μg was used. First, 5 μg of RNA was treated with Oligo dT (invitrogen, 18418-012) and reacted at 70° C. for 10 minutes to bind Oligo dT to RNA. 5x buffer solution, 10 mM dNTP, 0.1 M DTT, and Superscript II (Invitrogen, 18064-014, 200 U/μl) were added thereto, allowed to stand at room temperature for 10 minutes, and then reacted at 42°C for 50 minutes. After the reaction was completed, RNA was degraded by treating with RNaseH (Invitrogen, 18021-071) and reacting at 37° C. for 20 minutes. qPCR was performed with 2x Power SYBR Green (6 μL; Toyobo Life Science, Osaka, Japan) containing the obtained cDNA template (2 μL) and the following primers. 94 ℃ (15 seconds), 70 ℃ (60 seconds) was carried out 40 cycles, the primer for MUC2 (5'TCCTG TGGCA TCCAT GAAAC-3' primer for β-actin (5' CGCAG CTCAG TAACA GTCCG-3 ' was used. In addition, the reaction cycle in which the PCR product exceeded the fluorescence intensity threshold in the exponential step of PCR amplification was regarded as a threshold cycle (Ct). Cts were compared at constant fluorescence intensity. Expression of the gene of interest was quantified by comparison with β-actin.
그 결과 도 6과 같이, 위장관 운동이 저해된 Vehicle 그룹에서 뮤신 유전자의 발현은 감소하였지만, 본 발명의 감태 추출물을 처리한 실험군에서는 유의적으로 증가하였다. 그러므로 본 발명에 따른 감태 추출물은 뮤신 유전자의 발현을 촉진하는 효과를 가짐을 확인하였다.As a result, as shown in FIG. 6, the expression of mucin genes decreased in the Vehicle group in which gastrointestinal motility was inhibited, but significantly increased in the experimental group treated with the Ecklonia cava extract of the present invention. Therefore, it was confirmed that the Ecklonia cava extract according to the present invention has an effect of promoting mucin gene expression.
종합적으로 본 발명은 감태 추출물의 위장관 운동 촉진 효과를 확인한 것으로, 본 발명에 따른 감태 추출물은 위장관 운동이 저하된 동물에 투여하였을 때 위장관 운동을 촉진하였고 이와 관련된 호르몬의 분비를 촉진하였으며 뮤신의 분비를 촉진시키는 효과를 보였으므로, 이를 위장관 운동 촉진용 식품 조성물 및 사료 첨가용 조성물 등으로 유용하게 활용할 수 있다.Overall, the present invention confirms the gastrointestinal motility-accelerating effect of Ecklonia cava extract, and when administered to animals with reduced gastrointestinal motility, the Ecklonia cava extract according to the present invention promotes gastrointestinal motility, promotes the secretion of related hormones, and inhibits mucin secretion. Since it showed a stimulating effect, it can be usefully used as a food composition for promoting gastrointestinal motility and a composition for adding feed.
Claims (8)
상기 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food) 및 식품 첨가제(food additives)으로 이루어진 군에서 선택된 어느 하나인 것을 특징으로 하는, 식품 조성물.
According to claim 1,
The food composition is characterized in that any one selected from the group consisting of functional food (functional food), nutritional supplements (nutritional supplement), health food (health food) and food additives (food additives), food composition.
상기 감태 분말을 30 내지 70℃에서 12 내지 48시간 동안 순환 추출하는 추출 단계;
상기 추출된 감태 추출물을 여과하는 단계;
상기 여과된 감태 추출물을 건조 농축하는 단계; 및
상기 건조 농축된 감태 추출물을 용매에 희석하는 단계;를 포함하는 위장관 연동 운동(gastric motility) 작용 촉진용 식품 조성물의 제조 방법.Ecklonia pulverization step of producing Ecklonia cava powder by pulverizing Ecklonia cava;
Extraction step of circulating extraction of the Ecklonia cava powder at 30 to 70 ° C. for 12 to 48 hours;
Filtering the extracted Ecklonia cava extract;
Drying and concentrating the filtered Ecklonia cava extract; and
A method for producing a food composition for promoting gastric motility action, comprising diluting the dried and concentrated Ecklonia cava extract in a solvent.
상기 여과는 0.2 내지 0.6 ㎛의 여과 사이즈를 갖는 여과필터를 이용하여 수행하는 것을 특징으로 하는, 제조 방법.
According to claim 5,
Characterized in that the filtration is performed using a filter having a filtration size of 0.2 to 0.6 ㎛, manufacturing method.
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| Choi et al., Effects of Ecklonia cava as fucoidan-rich algae on growth performance, nutrient digestibility, intestinal morphology and caecal microflora in weanling pigs. Asian-Australas J Anim Sci. January 2017, Vol. 30, No. 1, pp. 64-70 1부.* |
| Song et al., Low molecular weight fucoidan ameliorating the chronic cisplatin-induced delayed gastrointestinal motility in rats. Food and Chemical Toxicology. 2012, Vol. 50, pp. 4468-4478 1부.* |
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