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Fish-elastin Hydrolysate: Development and Impact on the Skin and Blood Vessels
Abstract
This chapter introduces the development of enzymatic hydrolysate of fish elastin and also discusses its underlying mechanism. The skipjack bulbus arteriosus has been used for the production of elastin hydrolysate, due to the availability and abundance of resource. The chapter explains the impact of ingestion of the elastin hydrolysate in the skin and blood vessels. To explore the mechanism underlying the beneficial effects of skipjack-elastin hydrolysate, a food-derived elastin peptide in the human blood was screened. Consequently, Pro-Gly was identified. Pro-Gly enhances synthesis of elastin by human dermal fibroblast and the growth of human umbilical-vein endothelial cells. The results of the human trials and the occurrence of food-derived elastin peptide with biological activities in human blood indicate that fish-elastin hydrolysate is a promising food ingredient for improving skin and blood-vessel conditions.
... Elastin is an important protein component of the extracellular matrix in numerous tissues that require elasticity to perform their functions [35]. It is part of the core architecture that supports cell growth and, consequently, is in direct contact with multiple cell types. ...
... The next step in obtaining elastin is isolation. A standard method utilized is hot alkali extraction using 0.1 M NaOH at 98°C [35]. The disadvantage of this method is that incubating the tissues at high temperatures for over 50 min produces fragmented peptide products. ...
... Recently, various mammalian tissues have been used on an industrial scale to obtain an enzymatic hydrolysate of elastin. Since some studies suggest that this hydrolysate may enhance the appearance of the skin, marine sources of elastin, such as fish, have been proposed to satisfy consumer preference for products of this origin [35]. ...
The term biopolymer refers to materials obtained by chemically modifying natural biological substances or producing them through biotechnological processes. They are biodegradable, biocompatible, and non-toxic. Due to these advantages, biopolymers have wide applications in conventional cosmetics and new trends and have emerged as essential ingredients that function as rheological modifiers, emulsifiers, film-formers, moisturizers, hydrators, antimicrobials, and, more recently, materials with metabolic activity on skin. Developing approaches that exploit these features is a challenge for formulating skin, hair, and oral care products and dermatological formulations. This article presents an overview of the use of the principal biopolymers used in cosmetic formulations and describes their sources, recently derived structures, novel applications, and safety aspects of the use of these molecules.
... Collagen gives tissue mechanical strength, while elastin provides elasticity and plays a significant role in tissue development [3]. Only a few tissues, such as ligament and blood vessels are rich in elastin [4]. ...
... After enzymatic breakdown collagen and elastin form low molecular peptides, which are rapidly absorbed from the gastrointestinal tract and may act as signalling molecules, thereby stimulating anabolic processes in the organism [2,3,6]. The peptides with low molecular weight originated from collagen and elastin reveal the high biological activity [7,8]. ...
... High amounts of Gly and Pro residues play an important role in the biological activity of collagen [12]. Pro-Gly dipeptide originated from Skipjack-elastin hydrolysate enhances the elastin synthesis by human dermal fibroblast and the growth of human umbilical vein endothelial cells [3]. The biological activity of released peptides depends on the enzyme specificity and proteolysis conditions [12,13]. ...
... HC was used as both a nutraceutical and a cosmetic ingredient. The oral intake of a hydrolysate of EL showed beneficial effects on human skin, such as improvements in skin elasticity and wrinkles [21,53]. ...
The use of animal testing in the cosmetic industry is already prohibited in more than 40 countries, including those of the EU. The pressure for it to be banned worldwide in the future is increasing, so the need for animal alternatives is of great interest today. In addition, using animals and humans in scientific research is ethically reprehensible. This study aimed to prove some of the anti-aging properties of elastin (EL), hydrolyzed collagen (HC), and two vegan collagen-like products (Veg Col) in a tri-layered chitosan membrane that was ionically crosslinked with sodium tripolyphosphate (TPP). In the first approach, as a way of representing different layers of a biological system, such as the epidermis and the two dermis sublayers, EL, HC, or Veg Col were independently introduced into the two inner layers (2L(i+b)). Their effects were compared with those of their introduction into three layers (3L). Different experiments were performed on the membrane to test its elasticity, hydration, moisture retention, and pore reduction at different concentrations of EL, HC, and Veg Col, and the results were normalized vs. a blank membrane. This new alternative to animal or human testing can be suitable for proving certain efficacy claims for active ingredients or products in the pharmaceutical, nutritional, and cosmetic fields.
... In this regard, it is suggested that ingestion of elastin hydrolysate might improve the condition of the skin. Recent consumer preference for marine products, the availability and relatively low cost coupled with its demonstrated presence of elastin peptides and biological potency of skipjack (Shiratsuchi, Nakaba, Shigemura, Yamada, & Sato, 2013) makes it a good candidate for further exploring the mechanism of photoaging. We hypothesized that skipjack elastin hydrolysate small peptides have free radicals scavenging ability. ...
We aimed to identify skipjack elastin hydrolysate‐derived peptides, and explore its mechanism of photoprotection against ultraviolet A (UVA) induced skin damage. Two peptides, TGVLTVM (peptide A) and NHIINGW (peptide B), were successfully identified using Sephadex G‐15, analyzed and detected using LC‐MS/MS fitted with electrospray ionization (ESI‐MS/MS), and further synthesized using the Fmoc strategy. A UVA radiation‐induced damage cell model of Human keratinocytes cell line (HaCaT) was established and applied. We found that both peptides significantly reduced reactive oxygen species (ROS) generation, reversed mitochondrial membrane potential (MMP) by 12.40%–13.13% and 10.80%–12.90% and reduced cell apoptosis by 3.43%–6.44%, and 4.56%–6.03% for peptide A and B, respectively, compared to the UVA model group. In conclusion, peptides TGVLTVM and NHIINGW protected against UVA‐induced photoaging through the attenuation of oxidative stress, thus both peptides might be useful as promising ingredients in sunscreens or antiaging cosmetic products. Two peptides (TGVLTVM and NHIINGW) were purified and identified from the Skipjack elastin hydrolysate using sephadex G‐15 and LC/MS/MS. The two peptides exerted strong activity against UVA irradiation‐induced skin damage by scavenging intracellular ROS, reversed the loss of MMP, and reduced apoptosis in UVA irradiated HaCaT cell model compared to the control.
... Collagen is a fibrous connective-tissue protein [19] present in the extracellular space of bones, skins, tendons, cartilage and muscle [20]. It is the most abundant single protein present in fish [21], corresponding to 25% of total proteins in the vertebrates [22], [23], [24]. ...
Currently, several relevant industries incorporate bioactive fish molecules (proteins, lipids to minerals) in numerous products. The low prices and high quality of the raw material determine the use of these biomolecules. The high demand for fish originates an inadequate overexploitation of marine resources. In most cases, an industry-only uses part of the fish while the rest is discarded. Fish resources are finite and it is necessary to valorise all biomass in a sustainable way. The amounts of under-utilized residues generated by fish processing industries can create serious environmental problems and have led researchers and industries to actively seek for alternative strategies where the residues from fish transformation can be used as raw materials. The biorefinery concept in the search for sustainability is thriving with the use of all substrate to obtain products to be used by different industries whilst single product extraction ceases. Research has been focused on the use of innovative, economically and environmentally sustainable extraction methods to preserve the biological activity of the molecules and respond to the increasing awareness of consumers in product related issues. These pioneer methods can transform fish wastes into added-value by-products using an efficient and viable economic strategy. In this review, the extraction of different fish residues with environmentally friendly techniques for the obtention of different bioactive compounds will be addressed. The use of different residues and techniques to extract highly desirable biologically active compounds such as collagen, gelatin, lipids, and minerals will be reviewed, demonstrating the potentiality of this subject. The main goal of this review is to help researchers, policymakers and economic agents to understand the trends and the tools available to address such a relevant topic in the years to come.
Aims:
To determine the prophylactic effects of an elastin peptide derived from the bulbus arteriosus of bonitos and prolylglycine (PG), a degradation product of elastin peptide, on vascular dysfunction in spontaneously hypertensive rats (SHRs).
Main methods:
Male 15-week-old SHR/Izm rats were fed without (control group) or with elastin peptide (1 g/kg body weight) for 5 weeks (EP group), or were infused via an osmotic mini-pump for 4 weeks with PG (PG group) or saline (control group). Using thoracic aortas, we assessed endothelial changes by scanning electron microscopy. Vascular reactivity (contraction and relaxation) and pressure-induced distension was compared. mRNA production levels of endothelial nitric oxide synthase (eNOS) and intercellular adhesion molecule-1 (ICAM-1) were investigated by real-time-polymerase chain reaction.
Key findings:
Aortas of the EP group displayed limited endothelial damage compared with that in the control group. Under treatment of SHRs with elastin peptide, the effect of phenylephrine returned closer to the normal level observed in normotensive Wistar-Kyoto (WKY/Izm) rats. mRNA production of eNOS (but not ICAM-1) was greater in the EP group than in the control group. Endothelial damage was suppressed and pressure-induced vascular distension was greater in the PG group than in the corresponding control group.
Significance:
These results suggest that elastin peptide from bonitos elicits prophylactic affects hypertension-associated vascular dysfunction by targeting the eNOS signaling pathway. PG may be a key mediator of the beneficial effects of elastin peptide.
Video-enhanced microscopy was used to examine the interaction of elastin- or laminin-coated gold particles with elastin binding proteins on the surface of live cells. By visualizing the binding events in real time, it was possible to determine the specificity and avidity of ligand binding as well as to analyze the motion of the receptor-ligand complex in the plane of the plasma membrane. Although it was difficult to interpret the rates of binding and release rigorously because of the possibility for multiple interactions between particles and the cell surface, relative changes in binding have revealed important aspects of the regulation of affinity of ligand-receptor interaction in situ. Both elastin and laminin were found to compete for binding to the cell surface and lactose dramatically decreased the affinity of the receptor(s) for both elastin and laminin. These findings were supported by in vitro studies of the detergent-solubilized receptor. Further, immobilization of the ligand-receptor complexes through binding to the cytoskeleton dramatically decreased the ability of bound particles to leave the receptor. The changes in the kinetics of ligand-coated gold binding to living cells suggest that both laminin and elastin binding is inhibited by lactose and that attachment of receptor to the cytoskeleton increases its affinity for the ligand.
我々は新しい加速度脈波 (APG) の計測器を開発し, スコア法による血管の弾力性の評価を試みてきた。スコア法における“血管老化スコア”は正常な老化過程からどの程度逸脱しているのかを推測する指標である。今回の研究の目的は, 疾病の発症頻度ではなく, 疾病発症以前の動脈の老化性変化をAPGで捉えることによって, 動脈硬化関連要因の相対リスクを求め, APG検査の妥当性を検討することである。45～64歳の549名を対象にして検討した結果, スコア異常者の中では, 血圧異常と“喫煙中または喫煙経験あり”がスコア正常者よりも有意に多かった (35.4%, 67.7%) 。さらに, 10年前にも受診していた247名について検討すると, 中性脂肪高値群で10年後にスコア異常を示したのは22.8%と正常群に比べて有意に多く, 相対リスクは2.5倍であった (p<0.05) 。本研究で, 仮説として動脈硬化関連要因の中に含めた4, 000Hz聴力低下群でのスコア異常は22.9%, 肥満群でのスコア異常は18.6%, 喫煙者でのスコア異常は16.8%で, それぞれ正常群に比べて有意に多い傾向であり, 相対リスクはそれぞれ2.2倍, 1.8倍, 2.0倍であった (p<0.1) 。結果から以下のように結論された。1) 45～65歳の中高年のAPG検査において, 現状で高血圧者や喫煙者は動脈弾力性低下と判定されることが有意に多い。2) 35～55歳で高中性脂肪血症, 高音域聴力異常, 肥満, 喫煙などの要因を持つ者は, 10年後にはAPG検査において動脈弾力性低下と判定される可能性が高い。APGスコア法は, スクリーニングや患者の自覚を促す教育などに妥当性をもつと思われ, 生活習慣病の予防に一定の貢献をすることが期待される。さらに詳細な結果を得るためには, 今後はプロスペクティブなコホート・スタディが望まれる。
Peptides in the blood of subjects before and after collagen hydrolysate ingestion were fractionated by ion exchange and size-exclusion chromatographies and then derivatised with phenyl isothiocyanate. The derivatives were characterised by reserved phase (RP)-HPLC. Prolyl-hydroxyproline (Pro-Hyp), which has been identified in the previous studies, was detected as a major food-derived collagen peptide in the blood of all subjects (n=5). Another major peptide was identified as hydroxyprolyl-glycine (Hyp-Gly) in the blood of four subjects, which has not been detected in previous studies. The ratio of Hyp-Gly to Pro-Hyp depended on subjects and ranged from 0.00 to 5.04. Hyp-Gly was less susceptible to human serum peptidase than Pro-Hyp. Hyp-Gly enhanced the growth of mouse primary fibroblasts on collagen gels in a higher extent than Pro-Hyp. These findings suggest that Hyp-Gly plays a significant role in exerting the biological effects by ingestion of collagen hydrolysate.
ブタ大動脈よりエラスチンの酵素加水分解ペプチド（エラスチンペプチド）を調製し，その摂取が皮膚にもたらす影響について検討した．生後7週の雄ヘアレスマウスにエラスチンペプチドなどの試料を経口投与し，63日間の飼育を行った．皮膚老化モデルとして光老化モデルを採用し，週3回の頻度で紫外線を照射した．照射52日目および55日目にエラスチンペプチドとコラーゲンペプチドを混合投与した群およびコラーゲンペプチド投与群において，皮膚水分量が照射対照群に比べて有意に上昇し，エラスチンペプチドの単独投与群において上昇傾向が認められた．エラスチンペプチドやコラーゲンペプチドの単独投与に比べ，皮膚中の存在比やそれに近い比での混合投与による相乗効果が得られ，光老化に対する皮膚症状の改善が認められた．つまり，エラスチンペプチドがコラーゲンペプチドの摂取による皮膚水分量の上昇効果を増強することが示唆された．
Elastic fibres are an important component of the extracellular matrix and are made of two major components : the more abundant cross-linked elastic protein elastin and the multi-component microfibrils. The biosynthesis of elastic fibres is a complex process involving the interplay of many diverse proteins and genes with elastin as the major component. Tropoelastin is the soluble precursor of elastin and as such it plays a dominant role in elastogenesis. The expression of tropoelastin is under a complex control mechanism, with many isoforms existing. Numerous other components, including the microfibrillar proteins, the elastin-binding protein and lysyl oxidase, the enzyme which initiates elastin cross-linking, are involved in elastogenesis. Tropoelastin undergoes self-association under physiological conditions in a process referred to as coacervation, and this is thought to be a vital process during elastic fibre formation and in providing elasticity. Although various models explaining the elasticity of elastin have been put forward, only the fibrillar model is based on the coacervation ability of tropoelastin. With the molecular cloning of a number of components of the elastic fibre, the availability of these components is increasing and paves the way for in vitro modelling of complex interactions of the elastic fibre. This review emphasises the biochemistry of tropoelastin and its role in elastic fibre structure and assembly.
Elastin hydrolysate has apparent beneficial effects, and the food-derived peptide prolyl-glycine (Pro-Gly) is present in human blood after oral ingestion. Following ingestion of elastin hydrolysate (10 g/60 kg body weight) by healthy human volunteers, peripheral blood was used to prepare plasma samples from which peptides were extracted by solid phase extraction and fractionated by size-exclusion chromatography (SEC). Peptides in the SEC fractions were derivatized with phenyl isothiocyanate (PITC) and resolved by reversed phase (RP)-HPLC. Pro-Gly was the major food-derived elastin peptide, reaching a maximum (18 μM) at 30 min after ingestion, and decreasing to approximately 20% at 4 h after ingestion. Finally, in cell culture, levels of Pro-Gly in the medium above 0.1 μg/mL significantly enhanced elastin synthesis of normal human dermal fibroblasts (NHDF) without affecting the rate of cell proliferation.