KR102585342B1 - An anti-obesity composition comprising Lactobacillus plantarum HAC03 and Garcinia cambogia extract - Google Patents
An anti-obesity composition comprising Lactobacillus plantarum HAC03 and Garcinia cambogia extract Download PDFInfo
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- KR102585342B1 KR102585342B1 KR1020230017233A KR20230017233A KR102585342B1 KR 102585342 B1 KR102585342 B1 KR 102585342B1 KR 1020230017233 A KR1020230017233 A KR 1020230017233A KR 20230017233 A KR20230017233 A KR 20230017233A KR 102585342 B1 KR102585342 B1 KR 102585342B1
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- garcinia cambogia
- hac03
- lactobacillus plantarum
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
본 발명은 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물, 건강기능성 식품 조성물에 관한 것이다. 본 발명의 락토바실러스 플란타룸 HAC03 균주와 가르시니아 캄보지아 추출물의 조합은 체중과 지방 조직 무게를 감소시키고, 지방 세포 크기를 감소시키고, 지방합성을 저해하는 효능을 가짐으로써, 비만 질환의 치료제, 건강기능성 식품 등으로 활용할 수 있다.The present invention relates to a pharmaceutical composition for preventing or treating obesity and a health functional food composition containing Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract as active ingredients. The combination of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract of the present invention has the effect of reducing body weight and fat tissue weight, reducing fat cell size, and inhibiting fat synthesis, thereby providing treatment for obesity diseases and health functional properties. It can be used as food, etc.
Description
본 발명은 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물, 건강기능성 식품 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating obesity and a health functional food composition containing Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract as active ingredients.
비만은 비정상적이거나 과도한 지방 축적으로 인해 건강에 부정적인 영향을 미치는 질병이다. 비만은 WHO에서 지정한 10대 건강 위험 요소 중 하나이다. WHO에 따르면 2016년 19억 명(세계 인구의 25% 이상)이 과체중이고 그 중 6억 5천만 명 이상(세계 인구의 8.71%)이 비만에 해당한다. 비만은 단순히 체중이 증가하는 것뿐만 아니라, 제2형 당뇨병, 이상지질혈증, 고혈압, 지방간, 관상동맥질환, 대장암 등의 질병도 유발하며, 심리적 위축이나 우울증과 같은 심리적인 질병도 유발한다. Obesity is a disease that negatively affects health due to abnormal or excessive fat accumulation. Obesity is one of the top 10 health risk factors designated by WHO. According to WHO, in 2016, 1.9 billion people (more than 25% of the world's population) were overweight, and more than 650 million of them (8.71% of the world's population) were obese. Obesity not only causes weight gain, but also causes diseases such as type 2 diabetes, dyslipidemia, high blood pressure, fatty liver, coronary artery disease, and colon cancer, and also causes psychological diseases such as psychological atrophy and depression.
2021년 미국 식품의약국(FDA)과 유럽의약청(EMA) 모두에서 승인한 세 가지 비만 치료제가 있다. 이는 리라글루타이드(liraglutide), 올리스타트(orlistat) 그리고 리라글루타이드와 부프로피온(bupropion)의 조합이다. 이 중 리라글루타이드는 Saxenda 제품명으로 알려져 있으며, 2014년 FDA 승인, 2015년 EMA 승인, 2017년 한국 식품의약품안전처(MFDS) 승인을 받았다. 이후 리라글루타이드는 세계 비만치료제 시장 1위 자리를 지키고 있다. 리라글루타이드는 GLP-1 수용체에 결합하여 신호를 보내는 GLP-1(glucagon-like peptide-1)의 유사체다. GLP-1은 음식 섭취에 반응하여 신체에서 자연적으로 분비되는 호르몬으로써, 췌장에서 인슐린 분비를 촉진하여 과도한 혈당을 제거하고 글루카곤 분비를 낮추어 혈중 당분비를 조절한다. 이는 정상적인 혈당을 유지하는 중요한 역할을 하며 뇌의 수용체와 결합하여 시상하부로 신호를 전달함으로써 식욕을 감소시키고 억제한다. 그러나 설사, 구토 등의 부작용뿐만 아니라 간, 췌장 심장 등 다른 조직에도 부작용의 사례가 일부 보고되었다. 또한 유방암, 췌장암 등 일부 암의 발병률이 증가한 사례도 보고되었다. There are three obesity treatments approved by both the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) in 2021. These are liraglutide, orlistat, and a combination of liraglutide and bupropion. Among these, liraglutide is known under the product name Saxenda, and was approved by the FDA in 2014, the EMA in 2015, and the Ministry of Food and Drug Safety (MFDS) of Korea in 2017. Since then, liraglutide has maintained its No. 1 position in the global obesity treatment market. Liraglutide is an analog of GLP-1 (glucagon-like peptide-1) that binds to the GLP-1 receptor and signals. GLP-1 is a hormone naturally secreted by the body in response to food intake. It promotes insulin secretion from the pancreas to remove excessive blood sugar and lowers glucagon secretion to regulate sugar secretion in the blood. It plays an important role in maintaining normal blood sugar levels and reduces and suppresses appetite by binding to receptors in the brain and transmitting signals to the hypothalamus. However, in addition to side effects such as diarrhea and vomiting, some cases of side effects have been reported in other tissues such as the liver, pancreas, and heart. Additionally, cases of increased incidence of some cancers, such as breast cancer and pancreatic cancer, have been reported.
특정 질병, 예를 들어 비만을 치료하거나 예방할 수 있는 물질을 발견하는 데 있어 가장 중요한 것은 질병의 중증도를 줄이는 의미 있는 효과를 보장하는 것이다. 하지만 이와 동시에 부작용이 없어야 한다. 리라글루타이드 치료제의 부작용 같은 한계를 극복하기 위해, 세계적으로 다양한 연구팀이 안전성이 높고 동시에 약효가 검증된 물질을 발굴하는 연구가 진행되고 있다. The most important thing in discovering a substance that can treat or prevent a specific disease, such as obesity, is to ensure a meaningful effect in reducing the severity of the disease. But at the same time, there should be no side effects. In order to overcome limitations such as side effects of liraglutide treatment, various research teams around the world are conducting research to discover substances with high safety and proven efficacy.
세계적으로 식물을 기반으로 한 체중 조절은 고대부터 사용되어 온 방법이다. 그 중 가르시니아 캄보지아(Garcinia cambogia)는 전 세계적으로 많은 연구를 통해 항비만 효과 있는 것으로 밝혀져 이미 체중 조절 제품으로 사용되고 있다. 하이드록시시트릭산(Hydroxycitric acid, HCA)은 가르시니아의 체중 감량 기능의 핵심 성분으로 알려져 있다. HCA는 크게 두가지 방식으로 체중 감소에 영향을 미친다. 우선, HCA는 구연산과 유사한 구조를 가지고 있다. 설탕에서 지방을 만드는 과정에서 ATP-citrate lyase(ACL)는 시트르산(citric acid)과 결합하여 아세틸 CoA(acetyl CoA)를 형성하는데, 이 과정에서 HCA는 시트르산 대신 사용되어 이후의 콜레스테롤이나 지질의 합성을 억제한다. 또한, HCA는 뇌의 세로토닌 수치를 증가시켜 식욕을 억제하고 체중 감소에 영향을 미친다. 가르시니아 캄보지아는 안전성이 어느 정도 검증된 물질이이지만 일부 고농도로 섭취할 경우, 간 손상을 일으키는 부작용의 사례가 보고된 적이 있다. Around the world, plant-based weight control is a method that has been used since ancient times. Among them, Garcinia cambogia has been found to have anti-obesity effects through many studies around the world and is already being used as a weight control product. Hydroxycitric acid (HCA) is known to be a key ingredient in garcinia's weight loss function. HCA affects weight loss in two main ways. First of all, HCA has a structure similar to citric acid. In the process of making fat from sugar, ATP-citrate lyase (ACL) combines with citric acid to form acetyl CoA. In this process, HCA is used instead of citric acid to promote the subsequent synthesis of cholesterol or lipids. Suppress. Additionally, HCA increases serotonin levels in the brain, which suppresses appetite and affects weight loss. Garcinia cambogia is a substance that has been proven to be safe to some extent, but there have been reports of side effects causing liver damage when consumed in high concentrations.
본 발명자들은 항비만 효과를 나타내는 락토바실러스 플란타룸 HAC03 균주와 가르시니아 캄보지아 추출물을 복합 투여하는 경우, 락토바실러스 플란타룸 HAC03균주와 가르시니아 캄보지아 추출물과의 시너지 효과에 의해 항비만 효과가 현저하게 우수함을 확인함으로써 본 발명을 완성하였다.The present inventors found that when the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract, which exhibit anti-obesity effects, are administered in combination, the anti-obesity effect is significantly superior due to the synergistic effect between the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. By confirming this, the present invention was completed.
본 발명이 해결하고자 하는 과제는 비만 예방 또는 치료 효과를 가진 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 약학 조성물 또는 건강기능성 식품 조성물을 제공하는 것이다.The problem to be solved by the present invention is to provide a pharmaceutical composition or health functional food composition containing Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract as active ingredients, which have the effect of preventing or treating obesity.
상기의 과제를 해결하기 위해, 본 발명은 락토바실러스 플란타룸 HAC03 균주(수탁번호: KCTC 13242BP) 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물을 제공한다.In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating obesity containing Lactobacillus plantarum HAC03 strain (accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients.
상기 가르시니아 캄보지아 추출물은 추출물 총 중량 대비 하이드록시시트릭산(hydroxycitric acid, HCA)을 60~70중량%로 포함할 수 있다.The Garcinia cambogia extract may contain 60 to 70% by weight of hydroxycitric acid (HCA) based on the total weight of the extract.
상기 조성물은 알라닌 아미노전이효소(alanine aminotransferase), 아스파테이트아미노전이효소 (aspartate aminotransferase), 중성지방(triglyceride), 총 콜레스테롤(total cholesterol) 및 저밀도 지단백 콜레스테롤(Low density lipoprotein cholesterol)로 이루어진 군에서 선택된 1종 이상을 감소시킬 수 있다.The composition is one selected from the group consisting of alanine aminotransferase, aspartate aminotransferase, triglyceride, total cholesterol, and low density lipoprotein cholesterol. Species abnormalities can be reduced.
상기 조성물은 지방합성을 억제할 수 있다.The composition can inhibit fat synthesis.
또한, 본 발명은 락토바실러스 플란타룸 HAC03 균주(수탁번호: KCTC 13242BP) 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 개선용 건강기능성 식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving obesity containing Lactobacillus plantarum HAC03 strain (accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients.
상기 가르시니아 캄보지아 추출물은 추출물 총 중량 대비 하이드록시시트릭산(hydroxycitric acid, HCA)을 60~70중량%로 포함할 수 있다.The Garcinia cambogia extract may contain 60 to 70% by weight of hydroxycitric acid (HCA) based on the total weight of the extract.
본 발명의 락토바실러스 플란타룸 HAC03 균주와 가르시니아 캄보지아 추출물의 조합은 체중과 지방 조직 무게를 감소시키고, 지방 세포 크기를 감소시키고, 지방합성을 저해하는 효능을 가짐으로써, 비만 질환의 치료제, 건강기능성 식품 등으로 활용할 수 있다.The combination of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract of the present invention has the effect of reducing body weight and fat tissue weight, reducing fat cell size, and inhibiting fat synthesis, thereby providing treatment for obesity diseases and health functional properties. It can be used as food, etc.
도 1은 동물 실험 방법을 개략적으로 나타낸 것이다.
도 2 및 도 3은 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 체중 변화 감소를 확인한 것이다.
도 4는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 생화학 지표 변화를 확인한 것이다.
도 5는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 지방 조직 무게 변화를 확인한 것이다.
도 6 내지 도 9는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 지방 세포 크기 변화를 확인한 것이다. 도 6은 피하 지방 조직(subcutaneous adipose tissue, SAT), 도 7은 정소 지방 조직(epididymal adipose tissue, EAT), 도 8은 내장 지방 조직(mesenteric adipose tissue, MAT), 도 9는 갈색 지방 조직(brown adipose tissue, BAT)의 세포 크기 변화를 나타낸다.
도 10은 적출한 지방 조직에서 지방합성에 관여하는 사이토카인 변화를 확인한 것이다.Figure 1 schematically shows the animal experiment method.
Figures 2 and 3 confirm the decrease in body weight change when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
Figure 4 confirms changes in biochemical indicators when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
Figure 5 confirms the change in fat tissue weight when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
Figures 6 to 9 confirm changes in fat cell size when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. Figure 6 shows subcutaneous adipose tissue (SAT), Figure 7 shows epididymal adipose tissue (EAT), Figure 8 shows visceral adipose tissue (MAT), and Figure 9 shows brown adipose tissue (brown adipose tissue). Indicates changes in cell size of adipose tissue (BAT).
Figure 10 confirms changes in cytokines involved in fat synthesis in extracted fat tissue.
본 발명자들은 항비만 효과를 나타내는 락토바실러스 플란타룸 HAC03 균주와 가르시니아 캄보지아 추출물을 복합 투여하는 경우, 락토바실러스 플란타룸 HAC03균주와 가르시니아 캄보지아 추출물과의 시너지 효과에 의해 항비만 효과가 현저하게 우수함을 확인함으로써 본 발명을 완성하였다. 구체적으로 상기 균주와 가르시니아 캄보지아를 복합 투여하는 경우는, 균주 자체가 지닌 항비만 효과와 가르시니아 캄보지아 추출물이 나타내는 항비만 효과의 상승작용으로 균주 또는 가르시니아 캄보지아 추출물을 단독투여하는 경우보다 현저하게 우수한 항비만 효과가 나타남을 실험적으로 확인하였다. The present inventors found that when the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract, which exhibit anti-obesity effects, are administered in combination, the anti-obesity effect is significantly superior due to the synergistic effect between the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. By confirming this, the present invention was completed. Specifically, in the case of combined administration of the above strain and Garcinia cambogia, the anti-obesity effect is significantly better than when the strain or Garcinia cambogia extract is administered alone due to the synergistic effect of the anti-obesity effect of the strain itself and the anti-obesity effect of the Garcinia cambogia extract. It was experimentally confirmed that the effect occurred.
따라서 본 발명은 일 관점에서, 락토바실러스 플란타룸 HAC03 균주(수탁번호: KCTC 13242BP) 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물에 관한 것이다.Therefore, in one aspect, the present invention relates to a pharmaceutical composition for preventing or treating obesity containing Lactobacillus plantarum HAC03 strain (accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients.
본 발명의 락토바실러스 플란타룸(Lactobacillus plantarum)은 락토플란티바실러스 플란타룸(Lactoplantibacillus plantarum)과 동의어로 사용될 수 있다. Lactobacillus plantarum of the present invention can be used synonymously with Lactoplantibacillus plantarum .
상기 락토바실러스 플란타룸 HAC03(수탁번호: KCTC 13242BP) 균주는 백김치로부터 분리되었으며, 우수한 내산성 및 내담즙성을 보이며, 항생제 내성이 없고, 생체 아민 생성능이 없음을 확인한 바 있다(국내등록특허 제10-2266314호). 또한 상기 락토바실러스 플란타룸 HAC03 균주는 하기 서열번호 1로 표시되는 16s rDNA 서열을 포함하는 것을 특징으로 한다.The Lactobacillus plantarum HAC03 (accession number: KCTC 13242BP) strain was isolated from white kimchi and was confirmed to have excellent acid resistance and bile resistance, no antibiotic resistance, and no ability to produce biogenic amines (Domestic Registered Patent No. 10) -2266314). In addition, the Lactobacillus plantarum HAC03 strain is characterized by comprising a 16s rDNA sequence represented by SEQ ID NO: 1 below.
CTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGTCCGTCACAACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACC(SEQ ID NO: 1)
본 발명에서 가르시니아 캄보지아(Garcinia cambogia)는 인도와 동남아시아에서 자생하는 Guttiferae과의 열대성 과일나무이다.In the present invention, Garcinia cambogia is a tropical fruit tree of the Guttiferae family that grows natively in India and Southeast Asia.
본 발명에서 가르시니아 캄보지아 추출물은 가르시니아 캄보지아를 추출하여 수득한 추출물을 의미한다. 상기 가르시니아 캄보지아 추출물은 하이드록시시트릭산(Hydroxycitric acid, HCA)을 포함하고 있으며, HCA는 가르시니아 캄보지아 추출물의 항비만 효과의 핵심 성분이다. HCA는 콜레스테롤이나 지질의 합성을 억제하고, 뇌의 세로토닌 수치를 증가시켜 식욕을 억제함으로써 체중을 감소시키는 효과를 나타낸다.In the present invention, Garcinia cambogia extract refers to an extract obtained by extracting Garcinia cambogia. The Garcinia cambogia extract contains hydroxycitric acid (HCA), and HCA is a key ingredient in the anti-obesity effect of the Garcinia cambogia extract. HCA has the effect of reducing weight by suppressing appetite by inhibiting the synthesis of cholesterol or lipids and increasing serotonin levels in the brain.
본 발명에서 가르시니아 캄보지아 추출물은 추출물 종 중량 대비 50~70중량%로 HCA를 포함할 수 있으며, 바람직하게는 HCA를 60~70중량%로 포함할 수 있다. 본 발명의 일 실시예에서는 HCA를 65중량% 포함하는 가르시니아 캄보지아 추출물을 사용하였다. In the present invention, the Garcinia cambogia extract may contain 50 to 70% by weight of HCA relative to the weight of the extract species, and preferably may contain 60 to 70% by weight of HCA. In one example of the present invention, a Garcinia cambogia extract containing 65% by weight of HCA was used.
본 발명에서 용어"추출물"은 추출 처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다.In the present invention, the term "extract" refers to an extract obtained by extraction treatment, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a purified product of the extract, or a mixture thereof, and the extract itself and the extract formed using the extract. Includes extracts in all possible formulations.
또한, 본 발명의 가르시니아 캄보지아 추출물은 당업계에 공지된 통상의 방법에 따라, 즉 통상적인 온도와 압력의 조건 하에서, 통상적인 용매를 사용하여 제조될 수 있다. 바람직하게는 물, 탄소수 1~4의 무수 또는 함수 저급알코올 (예를 들면, 메탄올, 에탄올, 프로판올 및 부탄올), 프로필렌글리콜, 1,3 - 부틸렌글리콜, 글리세린, 아세톤, 디에틸에테르, 에틸 아세테이트, 부틸아세테이트, 디클로로메탄, 클로로포름, 핵산 및 이들의 혼합물로 구성된 군으로부터 선택되는 용매를 이용한 용매 추출법; 이산화탄소에 의한 감압 및 고온에 의한 추출법; 또는 초음파 추출법을 이용하여 추출한다. 바람직하게는 에탄올 또는 물을 이용하여 추출한 것이고, 더 바람직하게는 70%(v/v) 에탄올 또는 물을 이용하여 추출한 것이다.In addition, the Garcinia cambogia extract of the present invention can be prepared according to conventional methods known in the art, that is, under conventional conditions of temperature and pressure, and using conventional solvents. Preferably water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (e.g. methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate. , a solvent extraction method using a solvent selected from the group consisting of butyl acetate, dichloromethane, chloroform, nucleic acids, and mixtures thereof; Extraction method using reduced pressure and high temperature using carbon dioxide; Alternatively, it is extracted using an ultrasonic extraction method. Preferably, it is extracted using ethanol or water, and more preferably, it is extracted using 70% (v/v) ethanol or water.
본 발명의 가르시니아 캄보지아 추출물은 가르시니아 캄보지아 열매 추출물일 수 있으며, 바람직하게는 가르시니아 캄보지아 열매 껍질 추출물일 수 있다. The Garcinia cambogia extract of the present invention may be a Garcinia cambogia fruit extract, and preferably may be a Garcinia cambogia fruit peel extract.
상기 조성물은 체중 감소(실시예 1), 지방 조직 무게 감소(실시예 3), 지방 세포 크기 감소(실시예 4) 효과를 나타냄으로써 비만을 예방 또는 치료할 수 있음을 확인하였다.It was confirmed that the composition can prevent or treat obesity by exhibiting effects of reducing body weight (Example 1), reducing fat tissue weight (Example 3), and reducing fat cell size (Example 4).
상기 조성물은 알라닌 아미노전이효소(alanine aminotransferase), 아스파테이트아미노전이효소 (aspartate aminotransferase), 중성지방(triglyceride), 총 콜레스테롤(total cholesterol) 및 저밀도 지단백 콜레스테롤(Low density lipoprotein cholesterol)로 이루어진 군에서 선택된 1종 이상을 감소시킬 수 있다(실시예 2).The composition is one selected from the group consisting of alanine aminotransferase, aspartate aminotransferase, triglyceride, total cholesterol, and low density lipoprotein cholesterol. Species abnormalities can be reduced (Example 2).
또한, 상기 조성물은 지방합성을 억제할 수 있다. 구체적으로 본 발명의 일 실시예에서 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우, 지방합성에 관여하는 유전자인 FAS, ACC, PPARγ, SREBP1c의 발현이 감소된 것을 확인하였다(실시예 5).Additionally, the composition can inhibit fat synthesis. Specifically, in one embodiment of the present invention, when Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract were administered in combination, it was confirmed that the expression of FAS, ACC, PPARγ, and SREBP1c, genes involved in fat synthesis, was reduced (performed Example 5).
본 발명에서 용어 "예방"은 약학적 조성물의 투여로 질병을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.In the present invention, the term “prevention” refers to any action that suppresses a disease or delays its onset by administering a pharmaceutical composition.
본 발명에서 용어 "치료"는 질환의 진행을 끝내고(abrogating), 실질적으로 억제하고(inhibiting), 지연(slowing) 또는 역전(reversing)시키고, 질환의 임상학적 또는 심미적 증상을 실질적으로 개선하거나, 질환의 임상학적 또는 심미적 증상의 출현을 실질적으로 예방하는 것을 포함한다.As used herein, the term "treatment" refers to abolishing, substantially inhibiting, slowing or reversing the progression of a disease, substantially improving the clinical or aesthetic symptoms of a disease, or treating a disease. It includes substantially preventing the appearance of clinical or aesthetic symptoms.
상기 약학 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁제, 내용액제, 유제, 시럽제, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결 건조제 및 좌제로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있으며, 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.The pharmaceutical composition is any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried agents, and suppositories. It may have a dosage form, and may be in various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
본 발명에서 용어 "투여"는 어떠한 적절한 방법으로 대상에게 본 발명의 약학적 조성물을 도입하는 것을 말하며, 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있다.In the present invention, the term "administration" refers to introducing the pharmaceutical composition of the present invention to a subject by any appropriate method, and the administration route may be oral or parenteral, as long as it can reach the target tissue.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용될 수 있다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose or lactose, and gelatin. It can be prepared by mixing etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. may also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerogelatin, etc. can be used.
본 발명의 약학 조성물은 약학으로 유효한 양으로 투여할 수 있다. 본 발명에서 용어, "약학으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 개체 종류 및 중증도, 연령, 성별, 질병의 종류, 약물의 활성, 약물에 대한 민감도, 투여시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. In the present invention, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type and severity of the individual, age, gender, and type of disease. , can be determined based on factors including the activity of the drug, sensitivity to the drug, administration time, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the field of medicine. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and can be easily determined by a person skilled in the art.
본 발명의 약학 조성물은 비만 질환 치료를 목적으로 하는 개체이면 특별히 한정되지 않고, 어떠한 것이든 적용 가능하다. 예를 들면, 원숭이, 개, 고양이, 토끼, 모르모트, 랫트, 마우스, 소, 양, 돼지, 염소 등과 같은 비인간동물, 인간, 조류 및 어류 등 어느 것이나 사용할 수 있으며, 상기 약학 조성물은 비 경구, 피하, 복강 내, 폐 내 및 비강 내로 투여될 수 있고, 국부적 치료를 위해, 필요하다면 병변 내 투여를 포함하는 적합한 방법에 의하여 투여될 수 있다. 본 발명의 상기 약학적 조성물의 바람직한 투여량은 개체의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 예를 들어, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관 내 주사에 의해 투여될 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition of the present invention is not particularly limited and can be applied to any subject as long as it is intended for the treatment of obesity disease. For example, it can be used in any non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, humans, birds, and fish, and the pharmaceutical composition can be administered parenterally or subcutaneously. , may be administered intraperitoneally, intrapulmonaryly and intranasally, and for local treatment, if necessary, may be administered by any suitable method, including intralesional administration. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the individual's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. For example, it may be administered orally, rectally, or by intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection, but is not limited thereto.
적합한 총 1일 사용량은 올바른 의학적 판단범위 내에서 처치에 의해 결정될 수 있으며, 일반적으로 0.001 내지 1000 mg/kg의 양, 바람직하게는 0.05 내지 200 mg/kg, 보다 바람직하게는 0.1 내지 100 mg/kg의 양을 일일 1회 내지 수회로 나누어 투여할 수 있다.The appropriate total daily usage amount can be determined by treatment within the range of sound medical judgment, and is generally an amount of 0.001 to 1000 mg/kg, preferably 0.05 to 200 mg/kg, more preferably 0.1 to 100 mg/kg. The amount can be administered once or several times a day.
본 발명은 다른 관점에서, 락토바실러스 플란타룸 HAC03 균주(수탁번호: KCTC 13242BP) 및 가르시니아 캄보지아 추출물을 유효성분으로 포함하는 비만 예방 또는 개선용 건강기능성 식품 조성물에 관한 것이다.From another perspective, the present invention relates to a health functional food composition for preventing or improving obesity containing Lactobacillus plantarum HAC03 strain (accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients.
상기 식품 조성물이란 영양소를 한 가지 또는 그 이상 함유하고 있는 천연물 또는 가공품을 의미하며, 바람직하게는 어느 정도의 가공 공정을 거쳐 직접 먹을 수 있는 상태가 된 것을 의미하며, 통상적인 의미로서, 식품, 식품 첨가제, 건강기능식품 및 기능성 음료를 모두 포함하는 것을 말한다.The food composition refers to a natural product or processed product containing one or more nutrients, preferably in a state that can be eaten directly after a certain degree of processing, and in the usual sense, food, food. It includes all additives, health functional foods, and functional beverages.
본 발명의 식품 조성물은 환제, 분말, 과립, 침제, 정제, 캡슐 또는 액제 등의 형태를 포함할 수 있으며, 식품의 종류에는 별다른 제한이 없으며, 예를 들어 각종 음료, 껌, 차, 비타민 복합제, 건강보조 식품류 등이 있다.The food composition of the present invention may include the form of pills, powders, granules, infusions, tablets, capsules or liquids, and there is no particular limitation on the type of food, for example, various beverages, gum, tea, vitamin complexes, There are health supplements, etc.
상기 식품 조성물에는 다른 성분을 추가할 수 있으며, 그 종류는 특별히 제한되지 않는다. 예를 들어, 통상의 식품과 같이 여러 가지 생약 추출물, 식품학적으로 허용되는 식품보조첨가제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있으며, 이에 제한되지 않는다.Other ingredients can be added to the food composition, and their types are not particularly limited. For example, like regular foods, it may contain various herbal extracts, foodologically acceptable food additives, or natural carbohydrates as additional ingredients, but is not limited thereto.
본 발명에서 용어 "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.In the present invention, the term "food supplement" refers to a component that can be added to food as an auxiliary agent, and can be appropriately selected and used by a person skilled in the art as it is added to manufacture each type of health functional food. Examples of food supplements include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, and protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated drinks, etc., but the types of food supplements of the present invention are not limited to the above examples.
상기 천연 탄수화물의 예는 포도당, 과당 등의 단당류; 말토스, 수크로스 등의 이당류; 및 덱스트린, 시클로덱스트린 등의 다당류와, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이 있으며, 상기한 것 이외의 향미제로서 천연 향미제(타우마틴 등), 스테비아 추출물(레바우디오시드 Ａ, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다.Examples of the natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Flavoring agents other than those mentioned above include natural flavoring agents (such as thaumatin) and stevia extract (rebaudioside A, glycyrrhizin). hygin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be used to advantage.
본 발명의 식품 조성물에는 건강기능성 식품이 포함될 수 있다. 본 발명에서 용어 "건강기능성 식품"이란 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 기능성이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 건강기능성 식품은 당업계에서 통상적으로 사용되는 방법에 의하여 제조가능하며, 상기 제조시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. 또한 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어날 수 있다.The food composition of the present invention may include health functional foods. In the present invention, the term "health functional food" refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients with functional properties useful to the human body. Here, functionality means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes such as physiological effects. The health functional food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art. In addition, unlike general drugs, it is made from food, so it has the advantage of not having any side effects that may occur when taking the drug for a long time, and it can be highly portable.
일반적으로, 식품 조성물 중에 포함되는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물의 양은 전체 식품 중량의 0.1 내지 90 중량%로 가할 수 있다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있다. 또한, 본 발명에 따른 식품 조성물은 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물 이외에 본 발명이 목적으로 하는 주효과를 손상시키지 않는 범위 내에서 바람직하게는 주효과에 상승효과를 줄 수 있는 다른 성분을 함유하는 것도 무방하다.In general, the amount of Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract contained in the food composition may be 0.1 to 90% by weight of the total food weight. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range. In addition, the food composition according to the present invention contains, in addition to Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract, other ingredients that can preferably have a synergistic effect on the main effect within the range that does not impair the main effect aimed at by the present invention. It is also okay to contain .
이하, 실시예를 통하여 본 발명의 구성 및 효과를 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들 실시예에 의해 한정되는 것은 아니다.Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.
<실험방법><Experiment method>
1. 균주의 분리 및 동정1. Isolation and identification of strains
제조 후 10 ~ 15 일 정도 되어 발효가 진행중인 백김치를 구입한 후에 pH 를 측정하여 젖산 생산에 의한 pH 감소를 확인한 후 유산균을 분리하였다. 고압 멸균된 비닐백에 90 mL 생리식염수(0.85% NaCl/L)를 넣고, 백김치 샘플 10g을 넣어 5 분 간 200 rpm에서 균일하게 혼합하였다(Stomacher® 400 Circulator, Seward, UK). 1 ml를 분주하여 9 ml의 생리식염수(0.85% NaCl/L)에 10 배수로 계단 희석하여 MRS 고체배지(5.5% Lactobacillus MRS broth, BD Difco, USA 및 1.5% Bacteriological Agar, Affymetrix, USA)에 도말하였다. 그 후 37℃배양기에서 24 ~ 48시간 동안 키우고, 분리 및 동정하였다.After purchasing white kimchi that was undergoing fermentation about 10 to 15 days after production, the pH was measured to confirm the decrease in pH due to lactic acid production, and then the lactic acid bacteria were isolated. 90 mL of physiological saline (0.85% NaCl/L) was added to a high-pressure sterilized plastic bag, and 10 g of white kimchi sample was added and mixed uniformly at 200 rpm for 5 minutes (Stomacher ® 400 Circulator, Seward, UK). 1 ml was dispensed, serially diluted 10 times in 9 ml of physiological saline (0.85% NaCl/L), and plated on MRS solid medium (5.5% Lactobacillus MRS broth, BD Difco, USA and 1.5% Bacteriological Agar, Affymetrix, USA). . Afterwards, they were grown in an incubator at 37°C for 24 to 48 hours, isolated, and identified.
이후 자라난 균을 3% 과산화수소를 이용하여 37℃에서 24 ~ 48시간 동안 배양하면서 카탈라제 활성(catalase activity) 및 그람 염색 분석을 수행하였다. 카탈라제 음성 균주는 양방향 16S rDNA 유전자 염기서열을 분석을 통해 분류학적 종(species)을 규명하였다. 균주의 염기서열 규명은 ㈜ 솔젠트(한국)에 의뢰하여 수행하였다.Afterwards, the grown bacteria were cultured at 37°C for 24 to 48 hours using 3% hydrogen peroxide, and catalase activity and Gram stain analysis were performed. The taxonomic species of the catalase-negative strain was identified through bidirectional 16S rDNA gene sequence analysis. Identification of the nucleotide sequence of the strain was performed by requesting Solgent Co., Ltd. (Korea).
그 결과, 백김치로부터 분리한 본 발명의 균주는 카탈라제 음성이고, 그람 양성이며, 로드 모양인 균주로서 락토바실러스 플란타룸으로 확인되었고, 락토바실러스 플란타룸 HAC03으로 명명하였다. 해당 균주는 한국생명공학 연구원에 2020년 12월 28일 기탁하였다(수탁번호: KCTC 13242BP).As a result, the strain of the present invention isolated from white kimchi was identified as Lactobacillus Plantarum as a catalase-negative, Gram-positive, rod-shaped strain, and was named Lactobacillus Plantarum HAC03. The strain was deposited at the Korea Research Institute of Bioscience and Biotechnology on December 28, 2020 (Accession Number: KCTC 13242BP).
2. 동물 실험2. Animal testing
락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물의 비만 예방 및 개선 효과를 확인하기 위하여 동물 실험을 수행하였다. 도 1은 동물 실험 방법을 개략적으로 나타낸 것이다. 락토바실러스 플란타룸 HAC03 균주, 가르시니아 캄보지아 추출물 (HCA 65% 함유), 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 11주간 하루에 1회 구강 투여하였다. 락토바실러스 플란타룸 HAC03 균주 혼탁액을 1x108 CFU/mouse 농도로, 가르시니아 캄보지아 추출물은 200 mg/kg의 농도로 투여하였다. 시험 그룹은 하기 표 2와 같이 설정하였다.An animal experiment was performed to confirm the obesity prevention and improvement effects of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. Figure 1 schematically shows the animal experiment method. Lactobacillus plantarum HAC03 strain, Garcinia cambogia extract (containing 65% HCA), Lactobacillus plantarum HAC03 strain, and Garcinia cambogia extract were orally administered once a day for 11 weeks. Lactobacillus plantarum HAC03 strain suspension was administered at a concentration of 1x10 8 CFU/mouse, and Garcinia cambogia extract was administered at a concentration of 200 mg/kg. The test group was set as shown in Table 2 below.
일주일마다 그룹별로 증가한 체중을 기록하였다. 11주 후, 이산화탄소 가스로 안락사 시킨 후에 체중 및 조직 무게를 측정하였다. 또한, 심장에서 혈액을 추출하여, 혈장을 분리한 후 혈장에서 비만과 관련된 생화학지표를 측정하였다. 이 때 생화학 지표는 알라닌 아미노전이효소(alanine aminotransferase, ALT), 아스파테이트아미노전이효소 (aspartate aminotransferase, AST), 중성지방(triglyceride, TG), 총 콜레스테롤(total cholesterol, T-chol), 고밀도 지단백 콜레스테롤(high density lipoprotein cholesterol, HDL), 저밀도 지단백 콜레스테롤 (low density lipoprotein cholesterol, LDL)를 사용하였다.The weight gain for each group was recorded every week. After 11 weeks, the animals were euthanized with carbon dioxide gas and their body and tissue weights were measured. Additionally, blood was extracted from the heart, plasma was separated, and biochemical indicators related to obesity were measured in the plasma. At this time, the biochemical indicators are alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (T-chol), and high-density lipoprotein cholesterol. (high density lipoprotein cholesterol, HDL) and low density lipoprotein cholesterol (LDL) were used.
지방 조직을 적출하여 H&E 염색을 진행한 후, 지방세포의 크기 및 수를 측정하였다. Fat tissue was extracted, subjected to H&E staining, and the size and number of fat cells were measured.
3. 사이토카인 측정 3. Cytokine measurement
락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물의 비만 예방 및 개선 효과에 관여하는 분자 기전을 확인하기 위하여 지방 조직에서 RNA 동정 및 cDNA 합성을 한 후, qPCR을 통해 지방합성에 관련된 유전자의 발현량을 mRNA 레벨에서 측정하였다.To confirm the molecular mechanism involved in the obesity prevention and improvement effect of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract, RNA was identified and cDNA synthesized in adipose tissue, and the expression level of genes related to fat synthesis was measured through qPCR. Measured at the mRNA level.
구체적으로는 각 실험군의 마우스 대장을 균질화(homogenize)하고 Trizol®(RNA extraction buffer, Thermo Fisher)에 녹인 후, 제조사에서 제공한 프로토콜에 따라 RNA를 추출하였다. 추출한 RNA를 정량하고, 동일한 양의 RNA를 SuperiorScript III cDNA synthesis Kit (Enzynomics, Daejeon, South Korea)를 이용하여 역전사반응 (reverse transcription)을 일으켜 cDNA로 합성하였다. 이후, 표 3의 각 유전자 특이적인 프라이머, TOPreal qPCR 2X premix (SYBR Green with high ROX) (Enzynomics, Daejeon, South Korea) 및 cDNA를 이용하여 real-time PCR을 진행하였다. real-time PCR은 ABI 7500 Fast Real time PCR system (Applied Biosystems, Waltham, MA, USA)을 이용하였다. 상대적인 유전자 발현량은 △△Ct법을 사용하여 계산하였다. Specifically, the mouse colon of each experimental group was homogenized, dissolved in Trizol ® (RNA extraction buffer, Thermo Fisher), and RNA was extracted according to the protocol provided by the manufacturer. The extracted RNA was quantified, and the same amount of RNA was synthesized into cDNA by reverse transcription using the SuperiorScript III cDNA synthesis Kit (Enzynomics, Daejeon, South Korea). Afterwards, real-time PCR was performed using each gene-specific primer in Table 3, TOPreal qPCR 2X premix (SYBR Green with high ROX) (Enzynomics, Daejeon, South Korea), and cDNA. Real-time PCR was performed using the ABI 7500 Fast Real time PCR system (Applied Biosystems, Waltham, MA, USA). Relative gene expression levels were calculated using the △△Ct method.
(5'→ 3')Forward
(5'→ 3')
(5'→ 3')Reverse
(5'→ 3')
(서열번호 3)CATTTTCCTGAAGTTTCCGCAG
(SEQ ID NO: 3)
(서열번호 5)TGGAGAGCCCCACACACA
(SEQ ID NO: 5)
(서열번호 7)GCAGCAGGTTTGTCTTGGATGT
(SEQ ID NO: 7)
(서열번호 9)CAGCAGTGAGTCTGCCTTGAT
(SEQ ID NO: 9)
(서열번호 11)GGGGTGTTGAAGGTCTCAAAC
(SEQ ID NO: 11)
[실시예 1] 체중 변화 [Example 1] Weight change
도 2 및 도 3은 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 체중 변화 감소를 확인한 것이다.Figures 2 and 3 confirm the decrease in body weight change when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
그 결과, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹의 체중 증가량이 현저하게 감소하는 것을 확인하였다. 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹은 HFD 그룹에 비해 유의미한 체중 감소가 보이지 않았다(도 2 및 도 3).As a result, compared to the HFD group fed only a high-fat diet, it was confirmed that the weight gain of the group administered the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract in combination was significantly reduced. The group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone did not show significant weight loss compared to the HFD group (Figures 2 and 3).
[실시예 2] 생화학 지표 변화[Example 2] Changes in biochemical indicators
도 4는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 생화학 지표 변화를 확인한 것이다. Figure 4 confirms changes in biochemical indicators when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
생화학 지표는 알라닌 아미노전이효소(alanine aminotransferase, ALT), 아스파테이트아미노전이효소 (aspartate aminotransferase, AST), 중성지방(triglyceride, TG), 총 콜레스테롤(total cholesterol, T-chol), 고밀도 지단백 콜레스테롤(high density lipoprotein cholesterol, HDL), 저밀도 지단백 콜레스테롤(low density lipoprotein cholesterol, LDL)을 사용하였다.Biochemical indicators include alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (T-chol), and high-density lipoprotein cholesterol (high). density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) were used.
그 결과, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹에서 ALT와 AST 수치가 통계적으로 유의미한 감소한 것을 확인하였다. 또한, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹에서 TG, T-chol, LDL 수치가 통계적으로 유의미하게 감소한 것을 확인하였다.As a result, compared to the HFD group fed only a high-fat diet, a statistically significant decrease in ALT and AST levels was confirmed in the group administered the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract in combination. In addition, compared to the HFD group fed only a high-fat diet, a statistically significant decrease in TG, T-chol, and LDL levels was confirmed in the group administered combined Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
[실시예 3] 지방 조직 무게 변화[Example 3] Change in fat tissue weight
도 5는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 지방 조직 무게 변화를 확인한 것이다. 피하 지방 조직(subcutaneous adipose tissue, SAT), 정소 지방 조직(epididymal adipose tissue, EAT), 내장 지방 조직(mesenteric adipose tissue, MAT), 갈색 지방 조직(brown adipose tissue, BAT) 무게 변화를 살펴보았다.Figure 5 confirms the change in fat tissue weight when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. We examined changes in the weight of subcutaneous adipose tissue (SAT), epididymal adipose tissue (EAT), mesenteric adipose tissue (MAT), and brown adipose tissue (BAT).
그 결과, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹은 BAT에서만 유의미한 감소가 관찰되었으며 나머지 지방 조직에서는 유의미한 변화가 관찰되지 않았다. 반면, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹에서는 EAT를 제외한 SAT, MAT, BAT에서 유의미한 감소가 관찰되었다.As a result, compared to the HFD group fed only a high-fat diet, the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone showed a significant decrease only in BAT, and no significant changes were observed in the remaining fat tissues. On the other hand, in the group administered in combination with Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract, a significant decrease was observed in SAT, MAT, and BAT, excluding EAT.
[실시예 4] 지방 세포 크기 변화[Example 4] Change in fat cell size
도 6 내지 도 9는 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우의 지방 세포 크기 변화를 확인한 것이다. Figures 6 to 9 confirm changes in fat cell size when combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract.
도 6은 피하 지방 조직(subcutaneous adipose tissue, SAT)의 세포 크기 변화를 나타낸다. 도 6을 살펴보면, 고지방 식이만 먹인 HFD 그룹에 비해, 가르시니아 캄보지아 추출물을 단독으로 투여한 그룹과 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹 모두에서 피하 지방 조직의 크기가 감소한 것을 확인하였다. Figure 6 shows changes in cell size of subcutaneous adipose tissue (SAT). Looking at Figure 6, compared to the HFD group fed only a high-fat diet, the size of subcutaneous fat tissue decreased in both the group administered Garcinia cambogia extract alone and the group administered combined Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. Confirmed.
특히 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹보다 지방 조직의 크기가 더 크게 감소하였다.In particular, in the case of combined administration of Lactobacillus Plantarum HAC03 strain and Garcinia cambogia extract, the size of fat tissue was reduced more significantly than the group administered Lactobacillus Plantarum HAC03 strain or Garcinia cambogia extract alone.
도 7은 정소 지방 조직(epididymal adipose tissue, EAT)의 세포 크기 변화를 나타낸다. 도 7을 살펴보면, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 또는 복합 투여한 그룹 모두 통계적으로 유의미한 지방 세포 크기의 변화가 관찰되지는 않았다.Figure 7 shows changes in cell size of epididymal adipose tissue (EAT). Looking at Figure 7, compared to the HFD group fed only a high-fat diet, no statistically significant change in fat cell size was observed in either the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone or in combination.
도 8은 내장 지방 조직(mesenteric adipose tissue, MAT)의 세포 크기 변화를 나타낸다. 도 8을 살펴보면, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독으로 투여한 그룹과 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹 모두에서 내장 지방 조직의 크기가 감소한 것을 확인하였다. Figure 8 shows changes in cell size of visceral adipose tissue (MAT). Looking at Figure 8, compared to the HFD group fed only a high-fat diet, in both the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone and the group administered combined Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract It was confirmed that the size of visceral fat tissue was reduced.
특히 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹보다 내장 지방 조직의 크기가 유의미하게 감소하였다. In particular, when the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract were administered in combination, the size of visceral fat tissue was significantly reduced compared to the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone.
도 9는 갈색 지방 조직(brown adipose tissue, BAT)의 세포 크기 변화를 나타낸다. 도 9를 살펴보면, 고지방 식이만 먹인 HFD 그룹에 비해, 가르시니아 캄보지아 추출물을 단독으로 투여한 그룹과 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹에서 갈색 지방 조직의 크기가 감소한 것을 확인하였다. Figure 9 shows changes in cell size of brown adipose tissue (BAT). Looking at Figure 9, compared to the HFD group fed only a high-fat diet, the size of brown adipose tissue was confirmed to be reduced in the group administered Garcinia cambogia extract alone and the group administered combined Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract. did.
[실시예 5] 유전자 발현 변화[Example 5] Changes in gene expression
도 10은 적출한 지방 조직에서 사이토카인 변화를 확인한 것으로, A는 FAT Synthase(FAS), B는 Acetyl CoA Carboxylase(ACC), C는 Peroxisome proliferator- activated receptor gamma(PPARγ), D는 Sterol Regulatory Element Binding Protein 1 c(SREBP1c)의 발현량 변화를 나타낸 것이다. FAS, ACC, PPARγ, SREBP1c는 모두 지방합성에 관여하는 유전자다.Figure 10 shows changes in cytokines in extracted fat tissue, where A is FAT Synthase (FAS), B is Acetyl CoA Carboxylase (ACC), C is Peroxisome proliferator-activated receptor gamma (PPARγ), and D is Sterol Regulatory Element Binding. This shows the change in expression level of Protein 1 c (SREBP1c). FAS, ACC, PPARγ, and SREBP1c are all genes involved in fat synthesis.
구체적으로, 고지방 식이만 먹인 HFD 그룹에 비해, 저지방 식이를 먹인 LFD 그룹에서 FAS 발현량이 감소하는 것을 확인할 수 있다. 또한, 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 그룹에서 FAS 발현량이 감소하였다(도 10A).Specifically, compared to the HFD group fed only a high-fat diet, the amount of FAS expression was confirmed to be reduced in the LFD group fed a low-fat diet. In addition, compared to the HFD group fed only a high-fat diet, the amount of FAS expression was decreased in the group administered in combination with Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract (Figure 10A).
또한, 고지방 식이만 먹인 HFD 그룹에 비해, 저지방 식이를 먹인 LFD 그룹에서 ACC 발현량이 감소하는 것을 확인할 수 있다. 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독으로 먹인 그룹에서 ACC 발현량이 감소하였다. In addition, compared to the HFD group fed only a high-fat diet, it was confirmed that the ACC expression level decreased in the LFD group fed a low-fat diet. Compared to the HFD group fed only a high-fat diet, the ACC expression level decreased in the group fed only Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract.
특히 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹보다 ACC 발현량이 현저하게 감소하였다(도 10B). In particular, when the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract were administered in combination, the ACC expression level was significantly reduced compared to the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone (FIG. 10B).
또한, 고지방 식이만 먹인 HFD 그룹에 비해, 저지방 식이를 먹인 LFD 그룹에서 PPARγ 발현량이 감소하는 것을 확인할 수 있다. 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독으로 먹인 그룹에서 PPARγ 발현량이 감소하였다. In addition, compared to the HFD group fed only a high-fat diet, the level of PPARγ expression was confirmed to be decreased in the LFD group fed a low-fat diet. Compared to the HFD group fed only a high-fat diet, the amount of PPARγ expression was decreased in the group fed only Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract.
특히 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹보다 PPARγ 발현량이 현저하게 감소하였다(도 10C).In particular, when the Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract were administered in combination, the amount of PPARγ expression was significantly reduced compared to the group administered Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract alone (Figure 10C).
또한, 고지방 식이만 먹인 HFD 그룹에 비해, 저지방 식이를 먹인 LFD 그룹에서 SREBP1c 발현량이 감소하는 것을 확인할 수 있다. 고지방 식이만 먹인 HFD 그룹에 비해, 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독으로 먹인 그룹에서 SREBP1c 발현량이 감소하였다. In addition, compared to the HFD group fed only a high-fat diet, the level of SREBP1c expression was confirmed to be decreased in the LFD group fed a low-fat diet. Compared to the HFD group fed only a high-fat diet, the expression level of SREBP1c was decreased in the group fed only Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract.
특히 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여한 경우는 락토바실러스 플란타룸 HAC03 균주 또는 가르시니아 캄보지아 추출물을 단독 투여한 그룹보다 SREBP1c 발현량이 현저하게 감소하였다(도 10D).In particular, in the case of combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract, the amount of SREBP1c expression was significantly reduced compared to the group administered alone with Lactobacillus plantarum HAC03 strain or Garcinia cambogia extract (Figure 10D).
이를 통해, 락토바실러스 플란타룸 HAC03 균주 및 가르시니아 캄보지아 추출물을 복합 투여하는 경우는 효과적으로 비만을 치료 또는 예방할 수 있음을 확인하였다.Through this, it was confirmed that combined administration of Lactobacillus plantarum HAC03 strain and Garcinia cambogia extract can effectively treat or prevent obesity.
Claims (6)
상기 가르시니아 캄보지아 추출물은 추출물 총 중량 대비 하이드록시시트릭산 (hydroxycitric acid, HCA)을 60~70중량%로 포함하는 것을 특징으로 하는, 약학 조성물.
A pharmaceutical composition for preventing or treating obesity containing Lactobacillus plantarum HAC03 strain (Accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients,
A pharmaceutical composition, wherein the Garcinia cambogia extract contains 60 to 70% by weight of hydroxycitric acid (HCA) based on the total weight of the extract.
The method of claim 1, wherein the composition contains alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), and total cholesterol (T-chol). and low density lipoprotein cholesterol (LDL). A pharmaceutical composition characterized in that it reduces one or more types selected from the group consisting of low density lipoprotein cholesterol (LDL).
The pharmaceutical composition according to claim 1, wherein the composition inhibits fat synthesis.
상기 가르시니아 캄보지아 추출물은 추출물 총 중량 대비 하이드록시시트릭산 (hydroxycitric acid, HCA)을 60~70중량%로 포함하는 것을 특징으로 하는, 건강기능성 식품 조성물.A health functional food composition for preventing or improving obesity containing Lactobacillus plantarum HAC03 strain (Accession number: KCTC 13242BP) and Garcinia cambogia extract as active ingredients,
A health functional food composition, wherein the Garcinia cambogia extract contains 60 to 70% by weight of hydroxycitric acid (HCA) based on the total weight of the extract.
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| KR20210049395A (en) * | 2019-10-25 | 2021-05-06 | 농업회사법인 에스에스바이오팜 주식회사 | Composition Comprising Psyllium Husk, Garsinia Canbogia, and Probiotics and Functional Food Using the Same |
| KR102266314B1 (en) | 2020-12-30 | 2021-06-16 | 한동대학교 산학협력단 | Lactobacillus plantarum HAC03 and uses thereof |
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| KR20210049395A (en) * | 2019-10-25 | 2021-05-06 | 농업회사법인 에스에스바이오팜 주식회사 | Composition Comprising Psyllium Husk, Garsinia Canbogia, and Probiotics and Functional Food Using the Same |
| KR102266314B1 (en) | 2020-12-30 | 2021-06-16 | 한동대학교 산학협력단 | Lactobacillus plantarum HAC03 and uses thereof |
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