• 한국식품과학회지
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• 제50권4호
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• pp.420-429
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• 2018

본 연구에서는 식용 어류로부터 얻은 콜라겐을 효소 처리하여 글라이신-프롤린-하이드록시프롤린(Gly-Pro-Hyp, GPH)과 같은 트리펩타이드의 함량이 높은 콜라겐 가수분해물을 얻었고, 이 저분자 콜라겐 가수분해물(이하 콜라겐 트리펩타이드)의 피부 보습효과를 생체 외 실험(in vitro)과 생체 내 실험(in vivo)을 설계하여 증명하고자 하였다. 사람의 피부 섬유아세포 HDF 세포를 자외선 조사를 통해 인위적으로 손상된 세포를 만든 후, 콜라겐 트리펩타이드가 이를 회복시키는 정도를 몇 가지 보습인자의 활성과 발현량으로 확인하였다. Ceramide kinase의 효소활성은 콜라겐 트리펩타이드의 농도에 의존적으로 증가하였고, hyaluronic acid의 농도 역시 유의적으로 증가하였다. 더불어 히알루론산을 합성하는 유전자인 HAS2의 mRNA와 단백질 발현량이 시료의 처리양에 비례하여 증가하였고, 히알루론산을 분해하는 효소인 HYAL1의 수준은 콜라겐 트리펩다이드의 농도에 의존적으로 감소하였다. 상기의 효과는 1형 콜라겐을 합성하는 Collagen 1A 유전자의 단백질 발현량과 피부염증과 종양발생 시 증가한다고 알려진 CD44의 발현량의 확인으로 증명되었다. 유기용매의 도포로 피부건조를 유도한 동물모델을 사용하여 콜라겐 트리펩타이드의 피부에의 유효성을 평가하기 위해 경피수분손실량(TEWL)와 수분함유량을 직접적으로 측정한 바, 콜라겐 트리펩타이드 34 mg/kg bw의 고용량 처리군에서 가장 긍정적인 변화가 확인되었으나 17 mg/kg bw의 콜라겐 트리펩타이드 처리군에서도 수분함량에서 유의적인 변화가 관찰되었다. 이는 피부 건조로 유도된 가려움증에 대하여 동물이 자신의 몸을 긁는 행위를 계수한 결과와 같은 양상을 보였다. 또한 피부가 건조할 시 유도될 수 있는 염증인자인 $TNF-{\alpha}$와 IL-6의 단백질 발현량을 동물 피부 조직을 적출하여 확인하였고, 염색법을 통하여 피부 진피 내 탄력섬유의 변화를 가시적으로 제시하였다. 이 피부 조직에서의 Collagen 1A와 AQP3의 단백질 발현량 및 세라마이드 키나이제, HAS2, 그리고 HYAL1 효소활성 결과는 상기의 누적된 콜라겐 트리펩타이드의 보습 효과를 더불어 증명하여 주었다. 이상의 결과로부터 본 연구의 저자는 콜라겐 트리펩타이드가 피부 보습에 효능을 갖는 소재로서 활용가치가 높음을 알 수 있었으며, 현재 시장에서 유통되고 있는 분자량 1000 Da 이상의 콜라겐 가수분해물보다도 저분자화 되며 트리펩타이드를 다량 함유하는 상기의 소재가 증명된 유효성과 증가된 체내 흡수도에 근거하여 보다 확장된 가능성을 보여줄 수 있을 것이라 판단된다.

• Journal of Applied Biological Chemistry
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• 제62권1호
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• pp.101-107
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• 2019

The objectives of this study were to evaluate the anti-aging effects and investigate the effect of simulated gastrointestinal (GI) digestion on the anti-aging properties and intestinal permeation of the potential fish collagen hydrolysates (FCH). Therefore, procollagen synthesis, matrix metalloproteinase-1 (MMP-1) production, and Caco-2 cell permeability were analyzed before and after in vitro digestion for FCHs, low-molecular weight fractions (<1 kDa), and high molecular weight fractions (>1 kDa). After being subjected to GI digestion, the level of MMP-1 inhibition was maintained, although the procollagen production was significantly (>20%) lower with all samples. Also, the digested FCHs and their <1 kDa fraction yielded 9.1 and 13.8% increased peptide transport, respectively, compared to undigested samples. Based on the effective intestinal permeation and high digestive enzyme stability, the <1 kDa fraction of FCHs is a potential bioactive material suitable for anti-aging applications in the food and cosmetics industries.

• Journal of Microbiology and Biotechnology
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• 제34권2호
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• pp.415-424
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• 2024

This study reveals that low-molecular-weight collagen peptide (LMWCP) can stimulate the differentiation and the mineralization of MC3T3-E1 cells in vitro and attenuate the bone remodeling process in ovariectomized (OVX) Sprague-Dawley rats in vivo. Moreover, the assessed LMWCP increased the activity of alkaline phosphatase (ALP), synthesis of collagen, and mineralization in MC3T3-E1 cells. Additionally, mRNA levels of bone metabolism-related factors such as the collagen type I alpha 1 chain, osteocalcin (OCN), osterix, bone sialoprotein, and the Runt family-associated transcription factor 2 were increased in cells treated with 1,000 ㎍/ml of LMWCP. Furthermore, we demonstrated that critical bone morphometric parameters exhibited significant differences between the LMWCP (400 mg/kg)-receiving and vehicle-treated rat groups. Moreover, the expression of type I collagen and the activity of ALP were found to be higher in both the femur and lumbar vertebrae of OVX rats treated with LMWCP. Finally, the administration of LMWCP managed to alleviate osteogenic parameters such as the ALP activity and the levels of the bone alkaline phosphatase, the OCN, and the procollagen type 1 N-terminal propeptide in OVX rats. Thus, our findings suggest that LMWCP is a promising candidate for the development of food-based prevention strategies against osteoporosis.

• Preventive Nutrition and Food Science
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• 제18권2호
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• pp.124-132
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• 2013

In this study, novel peptides (NIPP-1, NIPP-2) derived from Navicula incerta (microalgae) protein hydrolysate were explored for their inhibitory effects on collagen release in hepatic fibrosis with the investigation of its underlying mechanism of action. TGF-${\beta}1$ activated fibrosis in LX-2 cells was examined in the presence or absence of purified peptides NIPP-1 and NIPP-2. Besides the mechanisms of liver cell injury, protective effects of NIPP-1 and NIPP-2 were studied to show the protective mechanism against TGF-${\beta}1$ stimulated fibrogenesis. Our results showed that the core protein of NIPP-1 peptide prevented fibril formation of type I collagen, elevated the MMP level and inhibited TIMP production in a dose-dependent manner. The treatment of NIPP-1 and NIPP-2 on TGF-${\beta}1$ induced LX-2 cells alleviated hepatic fibrosis. Moreover, ${\alpha}$-SMA, TIMPs, collagen and PDGF in the NIPP-1 treated groups were significantly decreased. Therefore, it could be suggested that NIPP-1 has potential to be used in anti-fibrosis treatment.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1401-1408
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• 2021

This study examined whether the oral administration of low-molecular-weight collagen peptide (LMCP) containing 3% Gly-Pro-Hyp with >15% tripeptide (Gly-X-Y) content could ameliorate osteoarthritis (OA) progression using a rabbit anterior cruciate ligament transection (ACLT) model of induced OA and chondrocytes isolated from a patient with OA. Oral LMCP administration (100 or 200 mg/kg/day) for 12 weeks ameliorated cartilage damage and reduced the loss of proteoglycan compared to the findings in the ACLT control group, resulting in dose-dependent (p < 0.05) improvements of the OARSI score in hematoxylin & eosin (H&E) and Safranin O staining. In micro-computed tomography analysis, LMCP also significantly (p < 0.05) suppressed the deterioration of the microstructure in tibial subchondral bone during OA progression. The elevation of IL-1β and IL-6 concentrations in synovial fluid following OA induction was dose-dependently (p < 0.05) reduced by LMCP treatment. Furthermore, immunohistochemistry illustrated that LMCP significantly (p < 0.05) upregulated type II collagen and downregulated matrix metalloproteinase-13 in cartilage tissue. Consistent with the in vivo results, LMCP significantly (p < 0.05) increased the mRNA expression of COL2A1 and ACAN in chondrocytes isolated from a patient with OA regardless of the conditions for IL-1β induction. These findings suggest that LMCP has potential as a therapeutic treatment for OA that stimulates cartilage regeneration.

• 한국축산식품학회지
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• 제35권2호
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• pp.156-163
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• 2015

The effects of low molecular-weight collagen peptides derived from porcine skin were investigated on the physicochemical and sensorial properties of chocolate ice cream. Collagen peptides less than 1 kDa in weight were obtained by sub-critical water hydrolysis at a temperature of $300^{\circ}C$ and a pressure of 80 bar. Ice cream was then prepared with gelatin powder and porcine skin hydrolysate (PSH) stabilizers mixed at seven different ratios (for a total of 0.5 wt%). There was no significant difference in color between the resulting ice cream mixtures. The increase in apparent viscosity and shear thinning of the ice cream was more moderate with PSH added than with gelatin. Moreover, the samples containing more than 0.2 wt% PSH had enhanced melting resistance, while the mixture with 0.2 wt% PSH had the lowest storage modulus at $-20^{\circ}C$ and the second highest loss modulus at 10℃, indicating that this combination of hydrocolloids leads to relatively softer and creamier chocolate ice cream. Among the seven types of ice creams tested, the mixture with 0.2 wt% PSH and 0.3 wt% gelatin had the best physicochemical properties. However, in sensory evaluations, the samples containing PSH had lower chocolate flavor scores and higher off-flavor scores than the sample prepared with just 0.5 wt% gelatin due to the strong off-flavor of PSH.

• Food Science and Biotechnology
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• 제17권2호
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• pp.241-246
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• 2008

A nutritional supplement was developed aiming at correcting the most common nutrient and caloric deficiencies encountered in elderly people (${\geq}60$ years old). The protein source was a mixture of whey protein isolates (WPI) and bovine collagen hydrolysate (BCH) with high nutritional and functional qualities making up 12% of the formulation. The carbohydrate fraction was composed of sucrose, inulin (soluble fiber), and fructo-oligosaccharide (prebiotic). The most commonly deficient essential minerals and vitamins were also included. Acceptance of the product was good according to both an elderly panel and a laboratory panel composing of both sexes and various ages. The stability of the formulations was evaluated and the estimated shelf life at room temperature (ca. $27^{\circ}C$) was approximately 4 months.

• 한국축산식품학회지
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• 제38권6호
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• pp.1144-1154
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• 2018

Both the calcium and collagen in bone powder are hard to be absorbed by the body. Although enzymatic hydrolysis by protease increased the bio-availability of bone powder, it was a meaningful try to further increase $Ca^{2+}$ release, oligopeptide formation and antioxidant activity of the sheep bone hydrolysate (SBH) by lactic acid bacteria (LAB) fermentation. Lactobacillus helveticus was selected as the starter for its highest protease-producing ability among 5 tested LAB strains. The content of liberated $Ca^{2+}$ was measured as the responsive value in the response surface methodology (RSM) for optimizing the fermenting parameters. When SBH (adjusted to pH 6.1) supplemented with 1.0% glucose was inoculated 3.0% L. helveticus and incubated for 29.4 h at $36^{\circ}C$, $Ca^{2+}$ content in the fermented SBH significantly increased (p<0.01), and so did the degree of hydrolysis and the obtaining rate of oligopeptide. The viable counts of L. helveticus reached to $1.1{\times}10^{10}CFU/mL$. Results of Pearson correlation analysis demonstrated that LAB viable counts, $Ca^{2+}$ levels, obtaining rates of oligopeptide and the yield of polypeptide were positively correlated with each other (p<0.01). The abilities of SBH to scavenge the free radicals of DPPH, OH and ABTS were also markedly enhanced after fermentation. In conclusion, L. helveticus fermentation can further boost the release of free $Ca^{2+}$ and oligopeptide, enhance the antioxidant ability of SBH. The L. helveticus fermented SBH can be developed as a novel functional dietary supplement product.

• 한국식품과학회지
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• 제40권4호
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• pp.436-442
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• 2008

본 연구를 통하여 피부 미용 소재로 알려져 있는 콜라겐 펩타이드를 섭취한 후 혈중에 나타나는 소화분해산물 중 콜라겐 특유의 아미노산인 hydroxyproline과 이것의 dipeptide 형태인 Pro-Hyp이 피부에 미치는 영향을 알아보고자 하였다. 피부 미용 소재를 경구 섭취하는 사람의 피부는 광노화 및 자연노화가 어느 정도 진행된 상태이므로, 이와 유사한 조건을 조성하기 위해 경구 투여를 시작하기 전에 hairless mouse에 7주간 주 3회 UV를 조사하여 주름을 선유발시켰다. 군 분리를 시킨 후 7주간 경구로 실험물질을 주 5회 반복 투여하면서, 주 3회 UV 조사를 병행하였으며, 시험 기간 중 임상증상, 체중변화를 관찰하였고, 3회 (0,4. 7주차) 피부주형을 채취하여 영상 분석을 실시하였으며, 2회 (0,7 주차)의 피부탄력측정 및 1회(7주차)의 TEWL측정을 병행하였다. 실험종료 후 피부조직의 병리조직학적 검사(H&E, Masson-Trichrome stain) 및 피부두께 측정을 실시하였다. 실험기간 중 이상 임상증상을 보이거나 사망한 개체는 없었으며, 체중에 대한 통계적으로 유의한 변화는 관찰할 수 없었다. 피부 육안관찰 시 NC군에 비해 UC군의 피부 주름 증가가 뚜렷하였으며, 시험군들은 UC군에 비해 주름 증가가 적었다. 경구 투여 0, 4, 7주차에 제작된 replica의 주름 지표를 통계 처리한 결과, UV/H군과 UV/P군은 DC군에 비해 감소경향은 보였으나 유의적인 차이는 없었다. 탄력측정검사 결과 UV를 쬐지 않은 NC군은 UC군에 비해 UA/UF, Ur/Ue, Ur/Uf 지표에서 유의적인 증가를 보였다. UV/H군과 UV/P군은 모두 0, 7주차 모두 유의적인 차이를 보이지 않았다. 피부 보습력 측정 결과 NC군 및 모든 실험군에서 UC군에 비해 유의적인 TEWL 감소를 관찰할 수 있었다. 피부 두께의 경우 NC군, UV/H군, UV/P군은 UC군에 비해 피부 두께가 유의적으로 감소하였다. 진피층에서의 교원질 확인을 위해 실시한 Masson-Trichrome stain의 경우 UC군에서는 상부 진피층에서의 염색상 소실이 관찰되었으나, 모든 실험군에서 진피층 손상이 거의 관찰되지 않아 실험물질에 의한 회복을 확인 할 수 있었다. 이상의 실험 결과 hydroxyproline, Pro-Hyp의 경구반복투여시 자외선에 의해 광노화가 유발된 피부에 보습 효과, 피부 두께 감소 효과, 진피층 손상 회복 효과를 유의적으로 확인할 수 있었으며, 단, 피부 주름 개선 정도는 통계적 유의성은 없이 경향성만을 확인할 수 있었다. Hydroxyproline과 이를 함유하는 dipeptide 형태의 Pro-Hyp를 경구 투여한 시험으로부터 확인한 피부 보습 및 진피층 회복 효과는 이 물질들이 유래된 콜라겐을 도포 또는 섭취했을 때 피부에 나타날 수 있는 작용들과 상통하며, 단독 미용 소재로도 활용될 수 있는 가능성을 보여주었다. 본 연구에서는 콜라겐 펩타이드를 섭취 시 체내에 흡수되어 혈액을 통해 각 조직에 운반되는 분해 산물이 피부에 미치는 영향을 동물 시험을 통해 살펴보았는데, 향후 연구에서는 관절, 혈관과 같은 다른 생체 조직들에 미치는 영향에 대해서도 검증해 볼 필요가 있다.

• 한국축산식품학회지
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• 제33권4호
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• pp.493-500
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• 2013

Gelatin is a collagen-containing thermohydrolytic substance commonly incorporated in cosmetic and pharmaceutical products. This study investigated the antioxidant activity of gelatin by using different reagents, such as 2,2-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS), 2,2-di (4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity-fluorescein (ORAC-FL) in a porcine gelatin hydrolysate obtained using gastrointestinal enzymes. Electrophoretic analysis of the gelatin hydrolysis products showed extensive degradation by pepsin and pancreatin, resulting in an increase in the peptide concentration (12.1 mg/mL). Antioxidant activity, as measured by ABTS, exhibited the highest values after 48-h incubation with pancreatin treatment after pepsin digestion. Similar effects were observed at 48 h incubation, that is, 61.5% for the DPPH assay and 69.3% for the ABTS assay. However, the gallic acid equivalent (GE) at 48 h was $87.8{\mu}M$, whereas $14.5{\mu}M$ GE was obtained using the ABTS and DPPH assays, indicating about sixfold increase. In the ORACFL assay, antioxidant activity corresponding to $45.7{\mu}M$ of trolox equivalent was found in the gelatin hydrolysate after 24 h hydrolysis with pancreatin treatment after pepsin digestion, whereas this activity decreased at 48 h. These antioxidant assay results showed that digestion of gelatin by gastrointestinal enzymes prevents oxidative damage.