KR102092865B1 - Geumgangsong extract with improved anti-inflammatory effect through plasma treatment and manufacturing method thereof - Google Patents
Geumgangsong extract with improved anti-inflammatory effect through plasma treatment and manufacturing method thereof Download PDFInfo
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- KR102092865B1 KR102092865B1 KR1020180099289A KR20180099289A KR102092865B1 KR 102092865 B1 KR102092865 B1 KR 102092865B1 KR 1020180099289 A KR1020180099289 A KR 1020180099289A KR 20180099289 A KR20180099289 A KR 20180099289A KR 102092865 B1 KR102092865 B1 KR 102092865B1
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- geumgangsong
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Abstract
본 발명은 플라즈마 처리를 통해 항염증 효과가 증진된 금강송 추출물 및 그의 제조방법에 관한 것으로, 플라즈마가 처리된 금강송 추출물은 LPS에 의해 증가된 NO의 생성을 억제하는 효과가 우수하므로, 플라즈마가 처리된 금강송 추출물을 유효성분으로 함유하는 본 발명의 조성물은 항염증용 건강기능식품, 염증성 질환의 치료제 또는 피부 염증 개선용 화장품의 소재로 유용하게 사용할 수 있다.The present invention relates to an extract of Geumgangsong with enhanced anti-inflammatory effect through plasma treatment and a method for manufacturing the same, and the plasma-treated Geumgangsong extract has an excellent effect of suppressing the production of increased NO by LPS. The composition of the present invention containing the extract of Geumgangsong as an active ingredient can be useful as an anti-inflammatory health functional food, a therapeutic agent for an inflammatory disease, or a cosmetic material for improving skin inflammation.
Description
본 발명은 플라즈마 처리를 통해 항염증 효과가 증진된 금강송 추출물 및 그의 제조방법에 관한 것이다.The present invention relates to an extract of Geumgangsong with improved anti-inflammatory effect through plasma treatment and a method for manufacturing the same.
염증(inflammation)은 물리적인 상처나 미생물에 감염되었을 때 일어나는 정상적인 생체의 방어기전(defense mechanism)의 일종이며, 이 염증작용을 통하여 발병요인(pathogen)을 중화시키거나 제거하고, 손상된 조직을 복구시켜 정상적인 구조와 기능을 하게 한다. 염증을 동반하는 대부분의 질환은 조직의 손상, 통증 및 가려움증과 같은 삶의 질을 떨어뜨리는 결과를 초래하고, 만성적인 염증상태는 관절염, 천식, 뇌와 척수의 다발성 경화증, 염증성 장 질환 및 동맥 경화증을 일으킨다.Inflammation is a type of defense mechanism of a normal living body that occurs when a person is injured by a physical wound or microorganism, and neutralizes or removes pathogens through this inflammatory action and repairs damaged tissue. Allows normal structure and function. Most diseases with inflammation result in a loss of quality of life such as tissue damage, pain and itching, and chronic inflammatory conditions include arthritis, asthma, multiple sclerosis of the brain and spinal cord, inflammatory bowel disease and atherosclerosis Causes
대식세포는 선천면역을 담당하는 주요 세포로, 사이토카인 및 박테리아 지질다당류 내독소(lipopolysaccharide, LPS) 같은 수많은 인자에 의해 활성화되고, 활성화된 대식세포는 산화질소(nitric oxide, NO) 및 프로스타글란딘 E2(prostaglandin E2, PGE2) 같은 염증 인자는 물론 TNF-α(tumor necrosis factor-α), IL-6(interleukin-6), IL-1(interleukin-1) 같은 전염증성 사이토카인을 생산한다. NO는 자유 라디칼(free radical)의 일종으로 산화질소 합성효소(nitric oxide synthase, NOS)에 의해 합성되며, eNOS(endothelial NOS), nNOS(neuronal NOS) 및 iNOS(inducible NOS)의 동형 단백질(isoform)이 존재한다(Cho 등. 2014. Biomol. Ther. 22: 288-294). IL-1, TNF-α, 인터페론-감마(interferon-gamma) 같은 전염증성 사이토카인들은 iNOS 발현을 유발하고 NO 생산을 일으킬 수 있다. 뇌암, 유방암, 폐암, 전립선암, 췌장암 및 흑색종 같은 많은 악성 암들이 iNOS 발현과 연관되어 있고, 과도한 NO의 생산은 상피세포암, 돌연변이 및 DNA 구조 손상을 일으킬 수 있다.Macrophages are the primary cells responsible for innate immunity, activated by numerous factors such as cytokines and bacterial lipopolysaccharide (LPS), and activated macrophages are nitric oxide (NO) and prostaglandin E 2 It produces inflammatory factors such as (prostaglandin E 2 , PGE 2 ) as well as pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1 (IL-1). NO is a free radical, and is synthesized by nitric oxide synthase (NOS), and isoforms of eNOS (endothelial NOS), nNOS (neuronal NOS) and iNOS (inducible NOS) Exists (Cho et al. 2014. Biomol. Ther. 22: 288-294). Pro-inflammatory cytokines such as IL-1, TNF-α, and interferon-gamma can cause iNOS expression and NO production. Many malignant cancers such as brain cancer, breast cancer, lung cancer, prostate cancer, pancreatic cancer and melanoma are associated with iNOS expression, and excessive NO production can lead to epithelial cancer, mutations and DNA structure damage.
한편, 금강송(Pinus densiflora for. erecta)의 서식 분포지역은 금강산 줄기의 계곡과 산복에서 태백산맥을 따라 경북 북부의 울진, 봉화군 일대와 강원 남부의 강릉, 삼척 등지에 넓게 분포하였으나, 벌목과 화전으로 크게 훼손돼 오늘날은 울진 소광리 일대에 국내 최대의 군락지가 형성되어 있다. Meanwhile, Geumgang Song ( Pinus densiflora for. The distribution area of erecta) was widely distributed in the valleys and mountains of the Geumgang Mountain Stem along the Taebaek Mountain Range in Uljin, Bonghwa-gun in the north of Gyeongbuk, and Gangneung and Samcheok in the southern part of Gangwon, but it was largely damaged by logging and fire warfare, and today it is in the area of Sogwang-ri, Uljin. Is the largest colony in Korea.
소나무는 한국, 일본, 만주 등에 자생하고 있고, 예로부터 잎, 솔방울, 꽃가루, 송진, 껍질 등 모든 부위가 구황식물로 이용되었다. 특히, 쉽게 채취할 수 있는 솔잎은 민간요법 등에 이용되어 고혈압, 당뇨병, 피부질환, 신경통 등의 성인병 치료와 불로장수를 위한 건강식품으로 이용되어 왔다. The pine tree is native to Korea, Japan, and Manchuria, and since ancient times, all parts such as leaves, pine cones, pollen, pines, and bark have been used as old plants. In particular, pine needles, which can be easily collected, have been used in folk medicine, etc., and have been used as health foods for the treatment of adult diseases such as hypertension, diabetes, skin diseases, and neuralgia and longevity.
현재 울진 지역의 관광지 조성과 육림 및 관리를 위해 지속적으로 이루어지는 간벌작업으로 인해 무한한 양의 금강송 간벌 부산물이 생산되고 있으며, 생산된 부산물은 대부분 퇴비나 톱밥제조 등에 사용되고 있다. 현재 송엽의 기능성에 대한 보고는 많이 있지만, 간벌 부산물인 송엽과 송절 혼합물의 기능성 연구는 활발하게 이루어지지 않아 기능성에 대한 과학적인 검증이 요구되고 있다. 또한, 불용자원인 간벌 부산물의 기능 연구를 통해 경제성이 우수한 기능성 식품 및 고부가가치 상품을 개발함으로써 새로운 소득원으로의 이용 가능성이 기대된다. Currently, infinite amounts of geumgangsong thinning by-products are being produced due to the ongoing thinning work to create tourist attractions, forestry and management in the Uljin region, and most of the produced by-products are used for composting or sawdust production. Currently, there are many reports on the functionality of pineal leaves, but the functional study of the pineal and pineal mixture, a byproduct of thinning, is not actively conducted, and scientific verification of the functionality is required. In addition, it is expected to develop functional foods and high value-added products with excellent economic feasibility through functional studies of thinning by-products, which are insoluble resources.
한편, 플라즈마란 초고온에서 음전하를 가진 전자와 양전하를 띤 이온으로 분리된 기체 상태를 말하며, 가장 저렴하게 플라즈마를 발생시킬 수 있는 방법은 전자파의 방전이다. 두 전극 사이에 전압을 걸어 전기장을 형성시킴으로써 플라즈마를 발생시키는 방식인데, 이때 발생된 플라즈마를 저온 플라즈마라고 한다. 플라즈마에는 전자, 양이온, 음이온, 자유라디칼 및 광자 등을 포함하는 활성산소종이 존재하며, 식품분야에서 주로 살균을 위해 사용되고 있지만, 본 발명자들은 이들이 물질의 구조를 변경시키고, 활성물질의 기능을 증진시킬 수 있다는 점에 착안하여 플라즈마 처리를 통해 우수한 효능소재인 금강송 추출물의 활성성분에 의한 효과를 더욱 증진시키고자 하였다.On the other hand, plasma refers to a gas state separated by electrons having a negative charge and positively charged ions at an extremely high temperature, and the most inexpensive method of generating plasma is discharge of electromagnetic waves. A plasma is generated by forming an electric field by applying a voltage between two electrodes, and the generated plasma is called a low-temperature plasma. Plasma contains active oxygen species including electrons, cations, anions, free radicals, and photons, and is mainly used for sterilization in the food field, but the present inventors have altered the structure of substances and promoted the function of active substances. In view of the fact that it is possible, it was intended to further enhance the effect of the active ingredient of the extract of Geumgangsong, an excellent efficacy material through plasma treatment.
한편, 한국공개특허 제2012-0077889호에는 송엽 또는 송절 추출물을 유효성분으로 포함하는 아토피 피부염 예방 또는 치료용 조성물이 개시되어 있지만, 본 발명의 플라즈마 처리를 통해 항염증 효과가 증진된 금강송 추출물 및 그의 제조방법에 대하여 개시된 바 없다. On the other hand, Korean Patent Publication No. 2012-0077889 discloses a composition for preventing or treating atopic dermatitis comprising a pineal or pineal extract as an active ingredient, however, an extract of Geumgangsong with improved anti-inflammatory effect through plasma treatment of the present invention and its No method has been disclosed.
본 발명은 상기와 같은 요구에 의해 도출된 것으로, 플라즈마 처리를 통해 항염증 효과가 증진된 금강송 추출물의 제조방법 및 이의 추출물을 유효성분으로 함유하는 항염증용 조성물을 제공하고, 상기 플라즈마 처리된 금강송 추출물이 LPS 처리에 의해 증가된 NO의 생성을 억제하는 것을 확인함으로써, 본 발명을 완성하였다. The present invention has been derived by the above-described needs, provides a method for preparing an extract of Geumgangsong with enhanced anti-inflammatory effect through plasma treatment, and an anti-inflammatory composition containing the extract thereof as an active ingredient, and the plasma-treated Geumgangsong The present invention was completed by confirming that the extract suppresses the production of increased NO by LPS treatment.
상기 과제를 해결하기 위하여, 본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 항염증용 건강기능식품 조성물을 제공한다. In order to solve the above problems, the present invention provides an anti-inflammatory health functional food composition containing a plasma-treated Geumgangsong extract as an active ingredient.
또한, 본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 염증성 질환의 예방 또는 치료용 약학 조성물을 제공한다. In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing the plasma-treated Geumgangsong extract as an active ingredient.
또한, 본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 피부 염증의 예방 또는 개선용 화장료 조성물을 제공한다. In addition, the present invention provides a cosmetic composition for preventing or improving skin inflammation containing plasma-treated Geumgangsong extract as an active ingredient.
또한, 본 발명은 In addition, the present invention
1) 금강송 건조 분말에 용매를 첨가하여 금강송 추출물을 획득한 후 동결건조하는 단계; 및1) obtaining a extract of Geumgangsong by adding a solvent to the dried powder of Geumgangsong, followed by freeze-drying; And
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 80~120 부피부의 물을 첨가한 후 플라즈마를 처리하는 단계;를 포함하는 항염효과가 증진된 금강송 추출물의 제조방법을 제공한다. 2) for 1 part by weight of the freeze-dried Geumgangsong extract of step 1), adding 80 to 120 parts by volume of water and then treating plasma; providing a method for producing Geumgangsong extract with enhanced anti-inflammatory effect do.
본 발명은 플라즈마 처리를 통해 항염증 효과가 증진된 금강송 추출물 및 그의 제조방법에 관한 것으로, 플라즈마가 처리된 금강송 추출물은 LPS에 의해 증가된 NO의 생성을 억제하는 효과가 우수하므로, 플라즈마가 처리된 금강송 추출물을 유효성분으로 함유하는 본 발명의 조성물은 항염증용 건강기능식품 조성물, 염증성 질환의 예방 또는 치료용 약학 조성물 또는 피부 염증의 예방 또는 개선용 화장료 조성물로 사용할 수 있다. The present invention relates to an extract of Geumgangsong with enhanced anti-inflammatory effect through plasma treatment and a method for manufacturing the same, and the plasma-treated Geumgangsong extract has an excellent effect of suppressing the production of increased NO by LPS. The composition of the present invention containing the extract of Geumgangsong as an active ingredient may be used as a health functional food composition for anti-inflammatory, a pharmaceutical composition for preventing or treating inflammatory diseases, or a cosmetic composition for preventing or improving skin inflammation.
도 1은 플라즈마가 처리된 금강송 추출물의 세포독성을 확인한 결과이다. CON은 음성 대조군이다.
도 2는 플라즈마 처리 시간에 따른 금강송 추출물의 NO 생성 억제효과를 농도별(25, 50㎍/㎖)로 확인한 결과이다. CON은 음성 대조군이고, LPS를 처리하여 NO의 생성을 유도하였다. ###은 control에 비해 LPS 처리군의 NO의 생성량이 유의미하게 증가하였다는 것을 의미하며, p<0.001이다. $$$은 플라즈마가 처리되지 않은 금강송 추출물 처리군의 NO의 생성량이 LPS 단독 처리군에 비해 유의미하게 감소하였다는 것을 의미하며, p<0.001이다. **, ***은 플라즈마를 처리하지 않은 금강송 추출물에 비해 플라즈마 처리된 금강송 추출물 처리군의 NO의 생성량이 유의미하게 감소하였다는 것을 의미하며, **은 p<0.01, ***은 p<0.001이다.1 is a result of confirming the cytotoxicity of the plasma-treated Geumgangsong extract. CON is a negative control.
2 is a result of confirming the inhibitory effect of NO production of Geumgangsong extract according to the plasma treatment time by concentration (25, 50㎍ / ㎖). CON is a negative control, and LPS was treated to induce NO production. ### means that the amount of NO produced in the LPS-treated group was significantly increased compared to the control, and p <0.001. $$$ means that the amount of NO produced in the plasma-treated Geumgangsong extract-treated group was significantly reduced compared to the LPS-only treatment group, and p <0.001. **, *** means that the amount of NO produced in the plasma-treated Geumgangsong extract treated group was significantly reduced compared to the plasma-treated Geumgangsong extract, ** is p <0.01, and *** is p < 0.001.
본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 항염증용 건강기능식품 조성물에 관한 것이다. The present invention relates to an anti-inflammatory health functional food composition containing a plasma-treated Geumgangsong extract as an active ingredient.
상기 금강송 추출물은 송엽(금강송의 잎) 및 송절(금강송의 가지와 줄기)의 혼합물로부터 추출한 것이 바람직하며, 더욱 바람직하게는 송엽 10 중량부에 대하여 송절 3~10 중량부로 혼합한 혼합물로부터 추출한 것이며, 더욱더 바람직하게는 송엽 10 중량부에 대하여 송절 5 중량부로 혼합한 혼합물로부터 추출한 것이지만, 이에 제한되는 것은 아니다. The extract of Geumgangsong is preferably extracted from a mixture of Songyeop (leaves of Geumgangsong) and Songjeol (branches and stems of Geumgangsong), more preferably extracted from a mixture of 3 to 10 parts by weight of Songjeol with respect to 10 parts by weight of Songye, Even more preferably, it is extracted from a mixture mixed with 5 parts by weight of pineapple with respect to 10 parts by weight of pine needle, but is not limited thereto.
본 발명의 일 구현 예에서, 상기 금강송 추출물의 용매는 C1~C4의 저급 알코올, 물 또는 이들의 혼합물인 것이 바람직하며, 더욱 바람직하게는 에탄올을 용매로 이용하는 것이지만, 이에 제한되지 않는다. In one embodiment of the present invention, the solvent of the extract of Geumgangsong is preferably C 1 to C 4 lower alcohol, water, or a mixture thereof, and more preferably, ethanol is used as a solvent, but is not limited thereto.
상기 플라즈마 처리는 텅스턴 전극 사이에 사인파 펄스형 고전압을 가하여 생성된 플라즈마를 금강송 추출물에 처리하는 것이지만, 이에 제한되는 것은 아니다. The plasma treatment is a plasma generated by applying a sine wave pulsed high voltage between tungsten electrodes, but is not limited thereto.
상기 고전압은 출력전압이 180~350V이고, 전류는 3~6A인 것이지만, 이에 한정하지 않는다.The high voltage has an output voltage of 180 to 350 V and a current of 3 to 6 A, but is not limited thereto.
상기 플라즈마 처리 시간은 80분 이상일 수 있으며, 바람직하게는 80~150분일 수 있으며, 더욱더 바람직하게는 80~130분일 수 있으나, 이에 제한되지 않는다.The plasma treatment time may be 80 minutes or more, preferably 80 to 150 minutes, and even more preferably 80 to 130 minutes, but is not limited thereto.
본 발명의 플라즈마는 대기압에서 두 전극 사이에 전압을 걸어 전기장을 형성시킴으로써 전자파의 방전을 통해 발생시키는 대기압 저온 플라즈마이다. The plasma of the present invention is an atmospheric pressure low-temperature plasma generated through discharge of electromagnetic waves by forming an electric field by applying a voltage between two electrodes at atmospheric pressure.
본 발명의 건강기능식품 조성물을 식품첨가물로 사용하는 경우, 상기 건강기능식품 조성물을 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 양은 그의 사용 목적(예방 또는 개선)에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시 본 발명의 건강기능식품 조성물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 그러나 건강을 목적으로 하는 장기간의 섭취인 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로 사용될 수 있다.When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used with other foods or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient can be appropriately used depending on the purpose of use (prevention or improvement). Generally, the health functional food composition of the present invention in the manufacture of a food or beverage is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on the raw material. However, in the case of long-term intake for health purposes, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
상기 건강기능식품의 종류에 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.There are no particular restrictions on the type of the health functional food. Examples of foods to which the health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum products, dairy products including ice cream, various soups, beverages, tea Drinks, alcoholic beverages, and vitamin complexes are included, and include all healthy foods in the ordinary sense.
또한, 본 발명의 건강기능식품 조성물은 식품, 특히 기능성 식품으로 제조될 수 있다. 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 포함할 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.In addition, the health functional food composition of the present invention may be prepared as a food, particularly a functional food. The functional food of the present invention includes ingredients that are commonly added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink agent, natural carbohydrate or flavoring agent may be included as an additional component in addition to the active ingredient. The natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrins, cyclodextrins, etc.) or sugar alcohols (eg. , Xylitol, sorbitol, erythritol, and the like). The flavoring agent may be a natural flavoring agent (eg, tau martin, stevia extract, etc.) and a synthetic flavoring agent (eg, saccharin, aspartame, etc.).
상기 건강기능식품 조성물 이외에 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 더 함유할 수 있다. 이러한 상기 첨가되는 성분의 비율은 크게 중요하진 않지만 본 발명의 건강기능식품 조성물 100 중량부에 대하여, 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.Various nutritional supplements, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid, in addition to the above-mentioned health functional food composition It may further contain a carbonation agent used in the beverage. The proportion of the components to be added is not particularly important, but it is generally selected from the range of 0.01 to 0.1 parts by weight with respect to 100 parts by weight of the health functional food composition of the present invention.
또한, 본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 염증성 질환의 예방 또는 치료용 약학 조성물에 관한 것이다. In addition, the present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing the plasma-treated Geumgangsong extract as an active ingredient.
본 발명에서 염증성 질환이란 관절염, 비염, 간염, 각막염, 위염, 장염, 신장염, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 요도염, 방광염, 화상 염증, 피부염, 치주염, 치은염 및 퇴행성 신경 염증 중에서 선택된 하나 이상인 것을 의미한다. In the present invention, the inflammatory disease is at least one selected from arthritis, rhinitis, hepatitis, keratitis, gastritis, enteritis, nephritis, bronchitis, pleurisy, peritonitis, spondylitis, pancreatitis, urethritis, cystitis, burn inflammation, dermatitis, periodontitis, gingivitis and degenerative nerve inflammation. Means
본 발명의 약학 조성물은 유효성분 이외에 약학적으로 허용되는 담체를 포함할 수 있으며, 이러한 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient, and such a carrier is commonly used in preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oil, and the like, but is not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components.
본 발명에 따른 약학 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. Suitable dosages of the pharmaceutical compositions according to the invention are variously prescribed by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity. Can be.
본 발명의 약학 조성물은 경구 또는 비경구로 투여할 수 있으며, 비경구 투여의 경우, 피부에 국소적으로 도포, 정맥 내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있다. The pharmaceutical composition of the present invention can be administered orally or parenterally, and for parenteral administration, it can be applied topically to the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and the like.
본 발명의 조성물에 포함되는 유효성분의 농도는 치료 목적, 환자의 상태, 필요기간 등을 고려하여 결정할 수 있으며, 특정 범위의 농도로 한정되지 않는다.The concentration of the active ingredient contained in the composition of the present invention may be determined in consideration of the purpose of treatment, the patient's condition, and the required period, and is not limited to a specific range of concentration.
본 발명의 약학조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 주사제, 크림, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제 중에서 선택된 어느 하나의 제형으로 제조될 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention pertains. It can be prepared by incorporating into a dose container. At this time, the formulation may be prepared in any one of a formulation selected from injections, creams, patches, sprays, ointments, warning agents, lotions, linen agents, pasta agents and cataplasmas, and may additionally include dispersants or stabilizers. have.
또한, 본 발명은 플라즈마 처리된 금강송 추출물을 유효성분으로 함유하는 피부 염증의 예방 또는 개선용 화장료 조성물에 관한 것이다. In addition, the present invention relates to a cosmetic composition for preventing or improving skin inflammation containing a plasma-treated Geumgangsong extract as an active ingredient.
본 발명의 일 구현 예에 따른 화장료 조성물에 있어서, 상기 피부 염증용 화장료 조성물은 스킨, 스킨 소프트너, 스킨 토너, 로션, 밀크로션, 모이스쳐로션, 영양로션, 마사지크림, 영양크림, 아이크림, 모이스쳐크림, 핸드크림, 에센스, 영양에센스, 팩, 클렌징폼, 클렌징워터, 클렌징크림, 바디로션, 바디클렌져, 비누 및 파우더 중에서 선택된 어느 하나의 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형으로 이루어진 화장료 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다. In the cosmetic composition according to an embodiment of the present invention, the cosmetic composition for skin inflammation is skin, skin softener, skin toner, lotion, milk lotion, moisturizer, nutrition lotion, massage cream, nutrition cream, eye cream, moisturizer Cream, hand cream, essence, nutrition essence, pack, cleansing foam, cleansing water, cleansing cream, body lotion, body cleanser, soap and powder may have any one selected from the formulation, but is not limited thereto. The cosmetic composition composed of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation, and the types and amounts of these components can be easily selected by those skilled in the art.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as carrier components Etc. can be used.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판-부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in the case of a spray, additionally, chlorofluorohydro Propellants such as carbon, propane-butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene Glycols, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.
본 발명의 화장료 조성물의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 아세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyltaurate, sarcosinate as a carrier component , Fatty acid amide ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like.
본 발명의 화장료 조성물은 형광물질, 살진균제, 굴수성 유발물질, 보습제, 방향제, 방향제 담체, 단백질, 용해화제, 당 유도체, 일광차단제, 비타민, 식물 추출물 등을 포함하는 부형제를 추가로 함유할 수 있다.The cosmetic composition of the present invention may further contain excipients including fluorescent materials, fungicides, oleogenic agents, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. .
또한, 본 발명은 In addition, the present invention
1) 금강송 건조 분말에 용매를 첨가하여 금강송 추출물을 획득한 후 동결건조하는 단계; 및1) obtaining a extract of Geumgangsong by adding a solvent to the dried powder of Geumgangsong, followed by freeze-drying; And
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 80~120 부피부의 물을 첨가한 후 플라즈마를 처리하는 단계;를 포함하는 항염효과가 증진된 금강송 추출물의 제조방법에 관한 것이다. 2) with respect to 1 part by weight of the freeze-dried Geumgangsong extract of step 1), 80 to 120 parts by volume of water, and then treating the plasma; relates to a method of manufacturing the Geumgangsong extract with enhanced anti-inflammatory effect will be.
본 발명의 일 구현 예에서, 상기 항염효과가 증진된 금강송 추출물은In one embodiment of the present invention, the anti-inflammatory effect is enhanced Geumgangsong extract
1) 금강송 건조 분말 1 중량부에 대하여 8~12 부피부의 70%(v/v) 에탄올을 첨가한 후, 실온에서 8~15시간 동안 진탕하여 금강송 추출물을 획득한 후 동결건조하는 단계; 및1) After adding 8% to 12 parts by volume of 70% (v / v) ethanol to 1 part by weight of Geumgangsong dry powder, shaking for 8-15 hours at room temperature to obtain Geumgangsong extract, followed by freeze-drying; And
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 80~120 부피부의 물을 첨가한 후 70~150분 동안 플라즈마를 처리하는 단계;를 포함하여 제조할 수 있으며, 2) to 1 part by weight of the lyophilized Geumgangsong extract of step 1), 80 to 120 parts by volume of water and then treating the plasma for 70 to 150 minutes; can be prepared, including,
바람직하게는Preferably
1) 금강송 건조 분말 1 중량부에 대하여 10 부피부의 70%(v/v) 에탄올을 첨가한 후, 실온에서 12시간 동안 진탕하여 금강송 추출물을 획득한 후 동결건조하는 단계; 및1) After adding 10 parts by volume of 70% (v / v) ethanol to 1 part by weight of Geumgangsong dry powder, shake it for 12 hours at room temperature to obtain a Geumgangsong extract, followed by freeze-drying; And
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 100 부피부의 물을 첨가한 후 80~130분 동안 플라즈마를 처리하는 단계;를 포함하여 제조하는 것이지만, 이에 한정하는 것은 아니다. 2) a step of treating plasma for 80 to 130 minutes after adding 100 parts by volume of water, based on 1 part by weight of the lyophilized Geumgangsong extract of step 1), but is not limited thereto. .
본 발명의 제조방법으로 제조된 금강송 추출물은 총 폴리페놀 함량이 일반 금강송 추출물에 비해 증가하고, 항산화 활성이 증진된다.
The extract of Geumgangsong prepared by the manufacturing method of the present invention increases the total polyphenol content compared to the general Geumgangsong extract and promotes antioxidant activity.
이하, 제조예 및 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 제조예 및 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.
Hereinafter, the present invention will be described in more detail by using manufacturing examples and examples. It is obvious to those skilled in the art that these production examples and examples are only intended to illustrate the present invention more specifically and that the scope of the present invention is not limited by them.
제조예Manufacturing example 1. One. 플라즈마plasma 처리된 금강송 추출물의 제조 Preparation of treated Geumgangsong extract
(1) 금강송 추출물의 제조(1) Preparation of Geumgangsong extract
송엽 및 송절이 2:1의 중량비로 혼합된 금강송 간벌 부산물을 건조하여 분쇄한 후, 상기 분쇄물 3g에 70%(v/v) 에탄올을 30㎖ 넣고, 상온에서 10시간 동안 진탕하며 추출한 후, 여과하여 동결건조하였다.
After drying and pulverizing the by-products of Geumgangsong mixed with pineapple and pineapple in a weight ratio of 2: 1, 30 ml of 70% (v / v) ethanol is added to 3 g of the pulverized product, and the mixture is shaken for 10 hours at room temperature and extracted It was filtered and lyophilized.
(2) 플라즈마 처리된 금강송 추출물의 제조(2) Preparation of plasma treated Geumgangsong extract
상기 동결건조한 금강송 에탄올 추출물 7g에 증류수 700㎖를 첨가하여 1%(w/v) 용액을 만든 후, 상기 용액을 플라즈마 발생기에 넣고, 일정시간(30, 60, 90 또는 120분) 동안 플라즈마를 처리하여 플라즈마 처리된 금강송 추출물을 제조하였다.After adding 700 ml of distilled water to 7 g of the lyophilized Geumgangsong ethanol extract to make a 1% (w / v) solution, put the solution in a plasma generator and process the plasma for a certain period of time (30, 60, 90 or 120 minutes). The plasma-treated extract of Geumgangsong was prepared.
본 발명에 사용한 플라즈마는 두 개의 텅스텐 전극 사이에 사인파 펄스형 고전압을 가하여 생성하였으며, 플라즈마 발생기는 전압/고주파 전원장치, 플라즈마 방전부, 처리 테이블의 3부분으로 구성된다. 플라즈마 방전부는 두께 5mm의 테프론 실드에 지름 3mm, 길이 20mm의 텅스텐 리드를 평행 배치한 후 매립하였다. 220V AC의 전원을 사용하였고, 출력전압은 300V, 전류는 5A로 고정하여 플라즈마를 생성하여 금강송 추출물에 처리하였다. The plasma used in the present invention was generated by applying a sine wave pulsed high voltage between two tungsten electrodes, and the plasma generator is composed of three parts: a voltage / high frequency power supply, a plasma discharge section, and a treatment table. The plasma discharge portion was embedded after placing a tungsten lead having a diameter of 3 mm and a length of 20 mm in parallel on a Teflon shield having a thickness of 5 mm. A power supply of 220V AC was used, the output voltage was fixed at 300V, and the current was fixed at 5A to generate plasma and treated with the extract of Geumgangsong.
본 발명의 비교예로는 상기와 동일한 방법으로 동결건조한 금강송 에탄올 추출물 7g에 증류수 700㎖를 첨가하여 1%(w/v) 용액을 만들어 플라즈마를 처리하지 않고 사용하였다.
As a comparative example of the present invention, 700 ml of distilled water was added to 7 g of ethanol extract of Geumgangsong lyophilized in the same manner as above to make a 1% (w / v) solution and used without plasma treatment.
실시예 1. 총 폴리페놀 함량 및 항산화능 측정Example 1. Measurement of total polyphenol content and antioxidant capacity
본 발명의 제조방법으로 제조된 플라즈마 처리된 금강송 추출물의 총 폴리페놀 함량 및 항산화능을 측정하였다.The total polyphenol content and antioxidant capacity of the plasma-treated Geumgangsong extract prepared by the production method of the present invention were measured.
총 폴리페놀 함량은 Gutfiger(1981)의 방법을 변형하여 측정하였으며, 제조예 1의 금강송 추출물의 희석액 100㎕에 폴린-시오칼투(Folin-Ciocalteau) 용액 100㎕를 가하고 3분간 정치한 다음 1㎖의 0.7M Na2CO3 용액을 가하고, 1시간 동안 반응시킨 후 분광광도계를 사용하여 750nm에서 흡광도를 측정하였다. 시료의 총 폴리페놀 함량은 탄닌산(tannic acid)을 이용하여 표준곡선을 작성한 후 계산하였다.The total polyphenol content was measured by modifying the method of Gutfiger (1981), 100 μl of a Folin-Ciocalteau solution was added to 100 μl of the dilution of the Geumgangsong extract of Preparation Example 1, and left standing for 3 minutes, and then 1 ml of After adding 0.7 M Na 2 CO 3 solution and reacting for 1 hour, absorbance was measured at 750 nm using a spectrophotometer. The total polyphenol content of the sample was calculated after preparing a standard curve using tannic acid.
ABTS 라디칼 소거활성은 Blois(1958)의 방법을 변형하여 측정하였다. 즉 7mM ABTS와 2.45mM 과황산칼륨(potassium persulfate) 용액을 혼합하여 하루 동안 암소에 방치하여 ABTS+·를 형성시킨 후, 상기 용액을 734nm에서 흡광도가 0.7~0.9이 되도록 증류수로 희석하였다. 그 후, 제조예 1의 금강송 추출물 50㎕에 희석된 ABTS 용액을 100㎕ 가한 후 5분간 반응시킨 다음 흡광도의 변화를 측정하였다. ABTS의 항산화능은 시료용액의 첨가구와 무첨가구의 흡광도 감소율로 계산하였다.ABTS radical scavenging activity was measured by modifying the method of Blois (1958). That is, 7 mM ABTS and 2.45 mM potassium persulfate (potassium persulfate) solution was mixed and left in the dark for 1 day to form ABTS + · , and then the solution was diluted with distilled water so that the absorbance at 734 nm was 0.7 to 0.9. Thereafter, 100 μl of ABTS solution diluted in 50 μl of Geumgangsong extract of Preparation Example 1 was added, reacted for 5 minutes, and then the change in absorbance was measured. The antioxidant capacity of ABTS was calculated by reducing the absorbance of the sample solution with and without additives.
플라즈마 처리에 의한 금강송 추출물의 총 폴리페놀 함량 및 ABTS 라디칼 소거능 변화는 표 1에 개시하였다. Changes in total polyphenol content and ABTS radical scavenging ability of Geumgangsong extract by plasma treatment are presented in Table 1.
총 폴리페놀 함량 및 항산화능은 30분 동안 플라즈마 처리한 금강송 추출물에서 플라즈마 처리 전에 비해 감소하는 경향을 나타내었으나, 금강송 추출물에 60분 이상 플라즈마를 처리한 경우 플라즈마 처리 전에 비해 총 폴리페놀 함량 및 항산화능이 증가하는 것을 확인할 수 있었다. The total polyphenol content and antioxidant capacity tended to decrease in plasma-treated Geumgangsong extract for 30 minutes compared to before plasma treatment. It was confirmed to increase.
(분)Plasma treatment
(minute)
(mg/추출물 1g)Total polyphenol content
(mg / extract 1g)
(100㎍/㎖)Radical scavenging capacity (%)
(100 μg / ml)
실시예Example 2. 세포생존율( 2. Cell viability ( cellcell viabilityviability ) 측정) Measure
제조예 1의 금강송 추출물이 세포생존에 미치는 영향을 확인하기 위하여 Mosmann(1983)의 방법을 이용하여 MTT(3-(4,5-dimeth-ylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) 분석을 실시하였다.MTT (3- (4,5-dimeth-ylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) using the method of Mosmann (1983) to confirm the effect of the extract of Geumgangsong of Preparation Example 1 on cell survival Analysis was conducted.
마우스 대식세포주인 raw 264.7 세포를 100㎕의 DMEM 배지가 담긴 96-웰 플레이트에 1일 동안 배양한 후, 제조예 1의 금강송 추출물(25 및 50㎍/㎖)을 처리하여 24시간 배양하였다. 그 후, 각 웰에 5㎎/㎖의 MTT 용액을 10㎕씩 분주한 후 3시간 동안 배양하면서 환원반응을 유도하였으며, 100㎕의 DMSO(dimethyl sulfoxide) 용액을 첨가하여 보라색의 포르마잔(formazan) 결정을 완전히 용해하였다. 그 후, 분광광도계를 이용하여 570㎚에서 흡광도를 측정하였으며, 금강송 추출물 무처리군(CON)의 생존율 100%를 기준으로 금강송 추출물 처리군의 상대적 세포 생존율을 계산하였다. The mouse macrophage cell line, raw 264.7 cells, was cultured in a 96-well plate containing 100 μl of DMEM medium for 1 day, and then treated with Geumgangsong extract (25 and 50 μg / ml) of Preparation Example 1 to incubate for 24 hours. Then, 10 μl of 5 mg / ml MTT solution was dispensed into each well, followed by incubation for 3 hours to induce a reduction reaction, and 100 μl of DMSO (dimethyl sulfoxide) solution was added to form purple formazan (formazan). Crystals were completely dissolved. Thereafter, the absorbance at 570 nm was measured using a spectrophotometer, and the relative cell viability of the Geumgangsong extract treated group was calculated based on the 100% survival rate of the Geumgangsong extract-free group (CON).
그 결과, 도 1에 나타난 바와 같이 제조예 1의 30, 60, 90 또는 120분 동안 플라즈마가 처리된 금강송 추출물 및 플라즈마가 처리되지 않은 금강송 추출물(0분)은 세포 생존율에 크게 영향을 주지 않은 것으로 보아, 본 발명의 금강송 추출물은 세포 독성이 거의 없다는 것을 알 수 있었다.
As a result, as shown in Figure 1, 30, 60, 90 or 120 minutes of Preparation Example 1, the plasma-treated Geumgangsong extract and the plasma-treated Geumgangsong extract (0 min) did not significantly affect cell viability. Boa, it was found that the extract of Geumgangsong of the present invention has little cytotoxicity.
실시예Example 3. 3. 플라즈마plasma 처리된 금강송 추출물의 Of treated Geumgangsong extract NONO (( nitricnitric oxideoxide ) 생성 억제효과) Inhibition effect
항염효과를 검증하기 위해 마우스 대식세포주인 RAW 264.7 세포를 96-웰 플레이트에 1×106cells/㎖가 되도록 접종하고, 37℃에서 5% CO2가 유지되는 배양기를 이용하여 24시간 동안 배양하였다. 이후 각 웰에 제조예 1의 금강송 추출물을 25 또는 50㎍/㎖의 농도로 1시간 동안 처리하고, 그 후 2㎍/㎖의 LPS를 처리하여 24시간 배양하였다. 배양 후 배양액의 상등액을 얻고, 상등액과 그리스(griess) 시약을 반응시킨 후, 분광광도계를 이용하여 540㎚ 파장에서 흡광도를 측정하였다. NO 생성율은 LPS 단독 처리군 대비 백분율로 나타내었다.In order to verify the anti-inflammatory effect, mouse macrophage cell line RAW 264.7 cells were inoculated in a 96-well plate to be 1 × 10 6 cells / ml, and 5% CO 2 was maintained at 37 ° C. Incubation was carried out for 24 hours using an incubator. Then, each well was treated with the extract of Geumgangsong of Preparation Example 1 at a concentration of 25 or 50 µg / ml for 1 hour, and then treated with 2 µg / ml of LPS to incubate for 24 hours. After cultivation, a supernatant of the culture solution was obtained, and the supernatant and the grease reagent were reacted, and absorbance was measured at a wavelength of 540 nm using a spectrophotometer. The NO production rate was expressed as a percentage compared to the LPS-only treatment group.
그 결과, 도 2에 나타난 바와 같이 금강송 추출물에 플라즈마를 60분 이상 처리하였을 경우 플라즈마가 처리되지 않은 금강송 추출물(0분)에 비해 NO 생성이 감소하는 것을 확인할 수 있었고, 특히 90분 이상 플라즈마를 처리한 금강송 추출물의 NO 생성 감소효과가 우수하였다. 하지만, 플라즈마를 30분 동안 처리한 금강송 추출물의 경우에는 플라즈마가 처리되지 않은 금강송 추출물에 비해 NO의 생성 억제효과가 나타나지 않았다. As a result, as shown in FIG. 2, when plasma was treated with the extract of Geumgangsong for 60 minutes or more, it was confirmed that NO generation was reduced compared to the extract of Geumgangsong (0 minute) where plasma was not treated, and plasma was particularly treated for 90 minutes or more. Han Geumgangsong extract had excellent NO reduction effect. However, in the case of the extract of Geumgangsong treated with plasma for 30 minutes, there was no effect of inhibiting the production of NO compared to the extract of Geumgangsong without plasma treatment.
따라서 플라즈마를 90분 이상 처리한 금강송 추출물은 NO의 생성 억제효과가 일반 금강송 추출물에 비해 증가함으로써, 우수한 항염증용 기능성 소재로 사용 가능하다는 것을 확인하였다. Therefore, it was confirmed that the Geumgangsong extract treated with plasma for 90 minutes or more can be used as an excellent anti-inflammatory functional material by increasing the inhibitory effect of NO compared to the general Geumgangsong extract.
Claims (10)
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 80~120 부피부의 물을 첨가한 후 80분~150분 동안 플라즈마를 처리하는 단계;를 포함하는 항염효과가 증진된 금강송 추출물의 제조방법.1) obtaining a extract of Geumgangsong by adding a solvent to the dried powder of Geumgangsong, followed by freeze-drying; And
2) for 1 part by weight of the lyophilized Geumgangsong extract of step 1), adding 80 to 120 parts by volume of water and then treating plasma for 80 minutes to 150 minutes; Geumgangsong with enhanced anti-inflammatory effect Method of manufacturing the extract.
1) 금강송 건조 분말 1 중량부에 대하여 8~12 부피부의 70%(v/v) 에탄올을 첨가한 후, 실온에서 8~15시간 동안 진탕하여 금강송 추출물을 획득한 후 동결건조하는 단계; 및
2) 상기 단계 1)의 동결건조된 금강송 추출물 1 중량부에 대하여, 80~120 부피부의 물을 첨가한 후 80분~150분 동안 플라즈마를 처리하는 단계;를 포함하여 제조하는 것을 특징으로 하는 항염효과가 증진된 금강송 추출물의 제조방법.The method of claim 9, wherein the anti-inflammatory effect is enhanced Geumgangsong extract
1) After adding 8% to 12 parts by volume of 70% (v / v) ethanol to 1 part by weight of Geumgangsong dry powder, shaking for 8-15 hours at room temperature to obtain Geumgangsong extract, followed by freeze-drying; And
2) with respect to 1 part by weight of the freeze-dried Geumgangsong extract of step 1), adding 80 to 120 parts by volume of water, and then treating plasma for 80 minutes to 150 minutes; Preparation method of extract of Geumgangsong with enhanced anti-inflammatory effect.
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