• Title/Summary/Keyword: Hyaluronidase inhibition assay

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Study on the Effect of Cimicifuga heracleifolia Ethanol Extract on Hyaluronic Acid Synthesis

  • Son, Hyun-Kyu;Ha, Hun-Yong
    • Applied Chemistry for Engineering
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    • v.33 no.6
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    • pp.557-562
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    • 2022
  • Hyaluronic acid (HA) is a mucopolysaccharide, occurring naturally in living organisms. It is one of the most hydrophilic molecules, so it has been known as being related to skin hydration and skin aging. The purpose of this study was to examine the effects of Cimicifuga heracleifolia ethanol extract on the hyaluronic acid synthesis and the inhibition of hyaluronidase activity. To determine cytotoxicity, hyaluronic acid synthase 2 (HAS2) gene expression, HA production and, hyaluronidase inhibitory effects, 3-(4,5-dimethylthiazol-2-ly)-2,5-diphenyl tetrazolium bromide (MTT) assay, real time - polymerase chain reaction (RT-PCR), hyaluronic acid enzyme linked immunosorbent assay (HA-ELISA), and hyaluronidase assay were used, respectively. When the Cimicifuga heracleifolia extract was treated in the HaCaT cells up to 500 ㎍/mL concentration, cytotoxicity was confirmed by the Cimicifuga heracleifolia extract at concentrations above 200 ㎍/mL. Therefore, the optimum concentration of all experiments used in this study was determined to be 200 ㎍/mL. HAS2 gene expression increased by Cimicifuga heracleifolia extract in a concentration-dependent manner at all treatment concentrations. The production rate of HA was tended to decrease at the highest concentration of 200 ㎍/mL. The hyaluronidase activity inhibition effect of Cimicifuga heracleifolia extract was very high compared to the control group. Based on these results, Cimicifuga heracleifolia extract was expected to have a moisturizing effect on human skin and special attention should be paid to the determination of the concentration of Cimicifuga heracleifolia when developing cosmetic materials using it.

Fourier Transform Ion Cyclotron Resonance (FT-ICR) MASS Spectrophotometric Analysis of Flower Petal from Paeonia lactiflora cv. ‘Red Charm’ and Evaluation of its Functional Activity (작약 레드참 꽃잎의 이온화원-푸리에 변환 질량분석과 기능성 연구)

  • Kim, June Hyun;Choi, Yong Bock;Lee, Ha Jung;Kim, Yong Hee;Kim, Jun Huan;Sim, Jung Min;Sohn, Young-Sun
    • Korean Journal of Plant Resources
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    • v.29 no.5
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    • pp.588-597
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    • 2016
  • Little attention has been paid to the functional aspect of the flower petal of Paeonia lactiflora, compared to that of its root. To determine the components of flower petal of Paeonia lactiflora, we conducted the Fourier transform ion cyclotron resonance (FT-ICR) MASS spectrophotometric analysis. We detected the 24 different types of ingredients from the 70% ethanol extracts of flower petal of peonia lactiflora cv. ‘Red Charm’. The main compounds were quercetin glucopyranosides, methyl gallate, paonioflolol and kaemperol glucopyranosides. We further tested its functional activity. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of the extracts was 87.9-90.4% at 0.1mg/ml. This result showed that these flower extracts have approximately 5-fold stronger antioxidant potential than a previous report with root extracts (Bang et al. 1999). The result of tyrosinase inhibition assay of Paeonia lactflora extract was almost similar to that of arbutin except significantly higher effect in the coral sunset extract at 0.1% concentration. Hyaluronidase inhibition assay showed 76.5% inhibition at 5% concentration of this flower extract, indicating that Peaonia lactiflora flower extracts have the major anti-inflammatory, anti-oxidant and brightening effects. Taken together, these results suggest these three Paeonia lactiflora species extracts might provide the basis to develop a new natural brightening agent.

Antioxidant and Anti-inflammatory Effects of Fractions from Pruni persicae Flos (도화(桃花, Pruni persicae Flos) 분획물의 항산화 및 항염증 효과)

  • Lee, Jin-Young;An, Bong-Jeun
    • The Korea Journal of Herbology
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    • v.27 no.5
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    • pp.55-63
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    • 2012
  • Objectives : The solvent extracts of Pruni persicae Flos were investigated for the activities of anti-oxidant and anti-inflammatory effects to apply as a functional ingredient for cosmetic products. Methods : In this study, the fractions of P. persicae Flos were extracted with 70.0% acetone and purified using Sephadex LH-20 column chromatography. As a result, eight fractions were isolated. We performed MTT assay, total polyphenol contents, DPPH free radical scavenging assay, SOD-like activity, xanthine oxidase inhibition assay, astringent activity assay, hyaluronidase inhibition assay and the production of nitric oxide. Results : For anti-oxidant effects, the electron donating ability of fraction (Fr.) 2-5, Fr.-8 isolated from P. persicae Flos was above 90.0% at 100 ppm respectively. The superoxide dismutase (SOD) - like activity of Fr.-5 isolated from P. persicae Flos was 92.1% at 1,000 ppm. The xanthine oxidase inhibitory effect of Fr.-6 isolated from P. persicae Flos was about 83.3% at 1,000 ppm. Hyaluronidase inhibition activity related to the anti-inflammation effect was 94.0% for Fr.-4 isolated from P. persicae Flos at 500 ppm. In the anti-inflammation effect, the Fr.-4 isolated from P. persicae Flos inhibited the generation of nitric oxide. Conclusions : All these findings suggested that the fractions of P. persicae Flos has a great potential as a cosmeceutical ingredient with a anti-oxidant and anti-inflammatory effects.

Anti-oxidant, Anti-aging, and Whitening Effects of Viscum album var. coloratum In Vitro (한국산 겨우살이의 항산화, 항노화 및 미백 효과)

  • Chang-Eui, Hong;Su-Yun, Lyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.4
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    • pp.295-302
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    • 2022
  • In this study, we investigated the anti-oxidant, anti-aging, and skin whitening effects of Korean mistletoe (Viscum album var. coloratum). The mistletoe fraction was composed of four types: hexane (HX), ethyl acetate (EA), butanol (BU), and methylene chloride (MC). In total phenol content assay, HX showed the highest phenol content among four fractions. In addition, EA significantly increased 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and catalase-like activities, and MC significantly increased superoxide dismutase (SOD)-like activity. When we compared IC50 value in the hyaluronidase and elastase inhibition assay, MC had the lowest IC50. In addition, we also performed tyrosinase inhibition assay to demonstrate the possibility of Korean mistletoe as a cosmetic component. HX showed the highest tyrosinase inhibition rate among the fractions.

Inhibitory effects of medicinal plants on elastase activity and biological activities in the active Plant extracts

  • Lee, Geon-Guk;Jo, Jung-Jae;Choe, Jung-Deok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.2
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    • pp.7-22
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    • 1997
  • Inhibitory effects of 200 medicinal plants on elastase activity were screened. Among the 200 plants, six plant extracts exhibited more than 65% of inhibition on elastase activity by their total methanol extracts at 1, 000$\mu\textrm{g}$/$m\ell$ as a final concentration. For six active plants including Areca catechu, Cinnamonum cassia, Myristica frograns, Curcumma longa, Alponia katsumadai, and Dryopteris crassirrhizoma, the inhibitory effects of their comsituents were examined on the activity of human leukocyte elastase, hyaluronidase and lipid peroxidation. In lipid peroxidation assay using TBA method, three of the 6 plants including Curcumma longa, Areca Catechu and Alponia katsumadai exhibited more than 70% of inhibition at the concentration of 1, 000$\mu\textrm{g}$/$m\ell$, but only one plant, Areca Ctechu showed high inhibitory effect on hyaluronidase activity. The results suggest that medicinal plants showing several biological activities may be potent inhibitors of anti-aging process on skin and that might be useful for application in cosmetics.

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Identification of Anti-oxidant, Anti-aging, and Whitening Effects of Diospyros kaki Extracts (감 추출물의 항산화, 항노화 및 미백 기능 규명)

  • Hong, Chang-Eui;Lyu, Su-Yun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.3
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    • pp.275-285
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    • 2022
  • In this study, we prepared two extracts using 1,3-butylene glycol (DBG) and ethanol (DET), with Diospyros kaki and determined the anti-oxidant, anti-aging, and whitening effects in vitro. Anti-oxidant activity was measured by DPPH and superoxide dismutase (SOD) method, and as a result, both DBG and DET extracts confirmed their anti-oxidant properties by exhibiting significant DPPH scavenging and SOD-like activities. For anti-aging activity, we measured elastase and hyaluronidase inhibition, and the inhibition of MMP-1 expression. Both DBG and DET significantly inhibited elastase and hyaluronidase activities dose-dependently, and MMP-1 expression was also reduced in both extracts. We also measured the whitening effects with tyrosinase activity and melanin production, but only DBG showed a decrease in tyrosinase. In summary, D. kaki extract has strong antioxidant, anti-aging, and whitening functions, and it is believed that it can be used as a cosmetic material in the future.

Clinical and molecular biological aspect of the hyaluronidases: basis and clinical overview for oriental medical application

  • Kim, Cheorl-Ho;Lee, Dong-Gyu;Jang, Jun-Hyouk;Kim, Jong-De;Nam, Kyung-Soo;Kim, Jeong-Joong;Park, Jong-Kun;Choo, Young-Kug;Kim, Hyung-Min;Lee, Young-Choon
    • Advances in Traditional Medicine
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    • v.1 no.1
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    • pp.8-27
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    • 2000
  • Components of extracellular matrix and the matrix-degrading enzymes are some of the key regulators of tumor metastasis and angiogenesis. Hyaluronic acid (HA), a matrix glycosaminoglycan, is known to promote tumor adhesion and migration, and its small fragments are angiogenic. Until now, we have compared levels of hyaluronidase, an enzyme that degrade HA, in normal adult prostate, benign prostate hyperplasia and prostate cancer tissues and in conditioned media from epithelial explant cultures, using a substrate (HA)-gel assay and ELISA-like assay (Kim et al., unpublished results). The present review described an overall characterization of hyaluronidases and its application to human diseases. The hyaluronidases are a family of enzymes that have, until recently, deed thorough explication. The substrate for these enzymes, hyaluronan, is becoming increasingly important, recognized now as a major participant in basic processes such as cell motility, wound healing, embryogenesis, and implicated in cancer progression. And in those lower life forms that torment human beings, hyaluronidase is associated with mechanisms of entry and spread, e.g. as a virulence factor for bacteria, for tissue dissection in gas gangrene, as a means of treponema spread in syphilis, and for penetration of skin and gut by nematode parasites. Hyaluronidase also comprises a component of the venom of a wide variety of organisms, including bees, wasps, hornets, spiders, scorpions, sh, snakes and lizards. Of particular interest is the homology between some of these venom hyaluronidases and the enzyme found in the plasma membrane of mammalian spermatozoa, attesting to the ancient nature of the conserved sequence, a 36% identity in a 300 amino acid stretch of the enzyme protein. Clearly, hyaluronidase is of biological interest, being involved in the pathophysiology of so many important' human disorders. Greater effort should be made in studying this family of enzymes that have, until recently, been overlooked. Also, oriental medical application of the hyaluronidase will be discussed with respect to inhibition and suppression of inflammation and malignacy.

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Characterization of Recombinant Bovine Sperm Hyaluronidase and Identification of an Important Asn-X-Ser/Thr Motif for Its Activity

  • Park, Chaeri;Kim, Young-Hyun;Lee, Sang-Rae;Park, Soojin;Jung, Yena;Lee, Youngjeon;Kim, Ji-Su;Eom, Taekil;Kim, Ju-Sung;Lee, Dong-Mok;Song, Bong-Suk;Sim, Bo-Woong;Kim, Sun-Uk;Chang, Kyu-Tae;Kim, Ekyune
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1547-1553
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    • 2018
  • Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.

Evaluation of Achyranthes japonica Ethanol Extraction on the Inhibition Effect of Hyluronidase and Lipoxygenase (쇠무릎 에탄올 추출물의 DPPH, 히알루로니다아제 및 리폭시게나아제 저해 효과)

  • Cho, Kyung-Soon
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1370-1376
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    • 2015
  • The 1, 1- diphenyl 2-picrylhyorazyl (DPPH) is a well-known radical and a trap (scavenger) for other radicals. Hyaluronidase (HAase) is an enzyme that depolymerizes the polysaccharide hyaluronic acid (HA) in the extracellular matrix of connective tissue. Lipoxygenase (LOX) enzyme was reported to convert the arachidonic, linoleic and other polyunsaturated fatty acid into biologically active metabolites involved in the inflammatory and immune responses. The purpose of the present study is to evaluate plant extracts as sources of natural antioxidants and to examine whether Achyranthes japonica having significant DPPH, HAase and LOX inhibitory activity. The inhibitory effect of HAase by A. japonica was assayed using a Morgan microplate assay. The antioxidant activity of the A. japonica extracts was measured on the basis of the scavenging activity of the stable 1, 1- diphenyl 2-picrylhyorazyl (DPPH) free radical. DPPH scavenging activity of matured roots of A. japonica was evaluated at 4.0 mg/ml was 87.8% and that of young roots was 86.2% at same concentration. The roots of A. japonica showed maximum inhibition of HAase activity (IC50 = 27.7 μg/ml). The highest LOX inhibition was recorded in the root extract among three vegetative parts. Inhibition of HAase activity of roots may contribute towards the development of herbal medicines. Although percent inhibition of lipoxygenase by Achyranthes japonica for all young and matured groups for leaves, stems, and roots at different concentrations, there were not show a statistically significant difference (p<0.05).

Screening of Hyaluronidase Inhibitory and Free Radical Scavenging Activity in vitro of Traditional Herbal Medicine Extracts (생약재 추출물의 hyaluronidase 저해 및 라디칼 소거 활성 검색)

  • 최수임;이윤미;허태련
    • KSBB Journal
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    • v.18 no.4
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    • pp.282-288
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    • 2003
  • For the screening of anti-inflammation and antioxidative activities, ethanolic extract of 40 species of traditional herbal medicines were examined their hyaluronidase inhibitory effect and radical scavenging activity in vitro. From the result of the hyaluronidase inhibitory activity using a Morgan-Elson assay, Astragali Radix, Eucommia Cortex, Schizandrae Fructus, Scutellaria Radix, Acanthopanacis Cortex, Chaenomelis Fructus, Amomum xanthioides Wallich and Moutan Radicis Cortex showed more than 50% hyaluronidase inhibitory effects at the concentration of 10 mg/mL. In the various solvent fractions (n-hexane, chloroform, ethyl acetate, water) prepared from ethanolic extracts, the ethyl acetate fraction of all extracts tested showed strong activity. Antioxidative activity was evaluated by assaying electron-donating ability to DPPH free radical and scavenging of hydroxyl radical (ㆍOH) generated through Fenton reaction, respectively. Rubus coreanus Miq, Moutan Radicis Cortex, Paeoniae Radix Alba, Plantaginis Semen and Sorbus commixta Hedl. showed high activity more than 90%, yet similar activity to $\alpha$-tocopherol and BHA at the concentration of 1 mg/mL in electron donating activity. The scavenging effects of ethanolic extracts on hydroxyl radical were investigated using a 2-deoxyribose oxidation method and tested all extracts showed significant radical scavenging activity. The experiment was also performed to examine whether herbal medicines having significant lipid peroxidation inhibitory activity, Schizandrae-Fructus is the strongest inhibitory activity in both linoleic acid and liposome peroxidation.