• Title/Summary/Keyword: HS68 Cells

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Convergence Study on Preparation of Anti-aging Peptides from Fish Collagen Hydrolysates (콜라겐 단백가수물을 이용한 항노화 펩타이드 제조에 대한 융합 연구)

  • Bae, In Young;Han, Yoo Kyung;Je, Hyun Jeong;Lee, Hyun Jun;Lee, Hyeon Gyu
    • Journal of the Korea Convergence Society
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    • v.11 no.5
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    • pp.61-72
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    • 2020
  • An anti-aging peptide was prepared from fish collagen hydrolysate (FCH) by ultrafiltration (MWCO; 1 kDa) and reversed-phased high performance liquid chromatography (RP-HPLC). Its anti-aging properties were evaluated based on the procollagen-synthesizing and MMP-1-inhibiting activities in Hs68 cells. A potent anti-aging peptide (fraction I-I) increased collagen synthesis by 46% and also inhibited MMP-1 secretion by 77%, compared with unpurified FCH. The amino acid sequence of fraction I-I was identified to be Gly-Arg-Arg-Gly-Asn-Lys (GRRGNK; the repeating Gly-X-Y sequence in collagen), and it had a molecular mass of 686.175 Da. It revealed that the anti-aging activity of GRRGNK was mainly due to skin protective effects. These results demonstrated that fish collagen hydrolysate may be a potential source of anti-aging peptides, which could be utilized in various field, including foods, cosmetics, and pharmaceuticals.

Biostability and Drug Delivery Efficiency of γ-Fe2O3 Nano-particles by Cytotoxicity Evaluation (세포독성 평가를 통한 γ-Fe2O3 나노입자의 생체안정성 및 약물전달효율)

  • Lee, Kwon-Jai;An, Jeung-Hee;Shin, Jae-Soo;Kim, Dong-Hee;Yoo, Hwa-Seung;Cho, Chong-Kwan
    • Korean Journal of Materials Research
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    • v.20 no.3
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    • pp.132-136
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    • 2010
  • This study examined the biostability and drug delivery efficiency of g-$Fe_2O_3$ magnetic nanoparticles (GMNs) by cytotoxicity tests using various tumor cell lines and normal cell lines. The GMNs, approximately 20 nm in diameter, were prepared using a chemical coprecipitation technique, and coated with two surfactants to obtain a water-based product. The particle size of the GMNs loaded on hangamdan drugs (HGMNs) measured 20-50 nm in diameter. The characteristics of the particles were examined by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-TEM) and Raman spectrometer. The Raman spectrum of the GMNs showed three broad bands at 274, 612 and $771\;cm^1$. A 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay showed that the GMNs were non-toxic against human brain cancer cells (SH-SY5Y, T98), human cervical cancer cells (Hela, Siha), human liver cancer cells (HepG2), breast cancer cells (MCF-7), colon cancer cells (CaCO2), human neural stem cells (F3), adult mencenchymal stem cells (B10), human kidney stem cells (HEK293 cell), human prostate cancer (Du 145, PC3) and normal human fibroblasts (HS 68) tested. However, HGMNs were cytotoxic at 69.99% against the DU145 prostate cancer cell, and at 34.37% in the Hela cell. These results indicate that the GMNs were biostable and the HGMNs served as effective drug delivery vehicles.

Protective Effect of Mulberry and Lithospermum erythrorhizon Extracts on Anti-aging against Photodamage (오디 및 자초추출물의 피부 광노화 보호효과)

  • Jeong, Yoo Seok;Jung, Hee Kyoung;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1744-1752
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    • 2013
  • We investigated the protective effect of UVB inducing photodamage from mulberry extract (ME) and Lithospermum erythrorhizon extract (LE). The contents of total anthocyanin and shikonin as a color compound of ME and LE were 4.92 mg/g and 9.58 mg/g, respectively. The electron donating ability and superoxide radical scavenging activity of ME were 84.32% and 76.34%, respectively. The oxygen radical absorbance capacity of the ME ($545.37{\mu}moles$ TE/g) was higher than LE ($427.18{\mu}moles$ TE/g). MMP-1 production in the HS68 cells were exposed to UVB suppressed by treatment with $200{\mu}g/mL$ of ME (68.6%) and LE (32.7%). ME and LE were applied to a skin aging mouse model, which was induced by the irradiation of UVB to the backs of hairless mice. The value of skin erythema index, wrinkle depth and thickness, epidermis thickness, and collagenous fiber damage in the experiment groups (MEL: ME 3%, MEM: ME 5%, MEH: ME 7%, LEL: LE 3%, LEM: LE 5%, LEH: LE 7%) were remarkably reduced than in the control group (only UVB exposure group), while water capacity increased. The level of total wrinkles depth in the skin was decreased to be 30% of the control group by MEH and LEM. These results suggest that ME and LE are useful cosmetic materials for skin protection against UVB-inducing.

Protective Effects of Fucoidan against UVB-Induced Oxidative Stress in Human Skin Fibroblasts (자외선에 의해 유도된 인간 피부 섬유아세포의 산화 스트레스에 대한 후코이단의 보호효과)

  • Ku, Mi-Jung;Lee, Myeong-Sook;Moon, Hee-Jung;Lee, Yong-Hwan
    • Journal of Life Science
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    • v.20 no.1
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    • pp.27-32
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    • 2010
  • Ultraviolet-B (UVB) radiation induces the formation of reactive oxygen species (ROS) and depletes stores of cellular antioxidants. Fucoidan, polysaccharides containing L-fucose and sulfate ester groups, are constituents of brown algae. In this study, the protective effects of fucoidan on UVB-induced oxidative stress in cultured human skin fibroblast HS68 cells were assessed. Pretreatment with fucoidan significantly reduced malondialdehyde (MDA) content in a dose-dependent manner. With fucoidan pretreatment at a dose of $100\;{\mu}g/ml$, the level of intracellular glutathione was increased by 21.5%, compared to UVB irradiation alone. Fucoidan significantly reduced UVB-induced ROS generation by 40.1% and 68.4% at 10 and $100\;{\mu}g/ml$, respectively, compared to UVB irradiation alone. The positive staining rates of senescence-associated $\beta$-galactosidase were reduced by 23.1% and 16.4% with 10 and $100\;{\mu}g/ml$ of fucoidan, compared to UVB irradiation alone. Fucoidan may exert a photoprotective effect against UVB radiation-induced oxidative stress.

Acceleration of heat shock-induced collagen breakdown in human dermal fibroblasts with knockdown of NF-E2-related factor 2

  • Park, Gunhyuk;Oh, Myung Sook
    • BMB Reports
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    • v.48 no.8
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    • pp.467-472
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    • 2015
  • Heat shock increases skin temperature during sun exposure and some evidence indicates that it may be involved in skin aging. The antioxidant response mediated by the transcription factor NF-E2-related factor 2 (Nrf2) is a critically important cellular defense mechanism that serves to limit skin aging. We investigated the effects of heat shock on collagenase expression when the antioxidant defense system was downregulated by knockdown of Nrf2. GSH and collagenases were analyzed, and the expression of inducible Nrf2, HO-1, and NQO1 was measured. HS68 cells were transfected with small interfering RNA against Nrf2. Heat shock induced the downregulation of Nrf2 in both the cytosol and nucleus and reduced the expression of HO-1, GSH, and NQO1. In addition, heat-exposed Nrf2-knockdown cells showed significantly increased levels of collagenase protein and decreased levels of procollagen. Our data suggest that Nrf2 plays an important role in protection against heat shock-induced collagen breakdown in skin. [BMB Reports 2015; 48(8): 467-472]

Glycosyl flavones from Humulus japonicus suppress MMP-1 production via decreasing oxidative stress in UVB irradiated human dermal fibroblasts

  • Nam, Eui Jeong;Yoo, Gyhye;Lee, Joo Young;Kim, Myungsuk;Jhin, Changho;Son, Yang-Ju;Kim, Sun Young;Jung, Sang Hoon;Nho, Chu Won
    • BMB Reports
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    • v.53 no.7
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    • pp.379-384
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    • 2020
  • Exposure to Ultraviolet (UV) light induces photoaging of skin, leading to wrinkles and sunburn. The perennial herb Humulus japonicus, widely distributed in Asia, is known to have anti-inflammatory, antimicrobial, and antioxidant effects. However, the physiological activities of isolated compounds from H. japonicus have rarely been investigated. This study focused on the isolation of active compounds from H. japonicus and the evaluation of their effects on photoaging in UVB-irradiated human fibroblast (Hs68) cells. When the extract and four fractions of H. japonicus were treated respectively in UVB-irradiated Hs68 cells to investigate anti-photoaging effects, the ethyl acetate (EtOAc) fraction showed the strongest inhibitory effect on MMP1 secretion. From EtOAc fraction, we isolated luteolin-8-C-glucoside (1), apigenin-8-C-glucoside (2), and luteolin-7-O-glucoside (3). These compounds suppressed UVB-induced MMP-1 production by inhibiting the phosphorylation of the mitogen-activated protein kinases (MAPKs) and activator protein-1 (AP-1). When the antioxidant activity of the compounds were estimated by conducting western blot, calculating the bond dissociation energies of the O-H bond (BDE) at different grade, and measuring radical scavenging activity, we found luteolin-8-C-glucoside (1) showed the strongest activity on the suppression of UVB-induced photoaging. These results demonstrate the inhibitory effect of three flavone glycosides derived from H. japonicus on MMP-1 production, MAPK and AP-1 signaling, and oxidative stress; this could prove useful in suppressing UVB induced photoaging.

Fermented Unpolished Black Rice (Oryza sativa L.) Inhibits Melanogenesis via ERK, p38, and AKT Phosphorylation in B16F10 Melanoma Cells

  • Sangkaew, Orrarat;Yompakdee, Chulee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1184-1194
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    • 2020
  • Melanin is a major factor that darkens skin color as one of the defense systems to prevent the harmful effects of UV light. However, darkened skin from the localized or systemic accumulation of melanin is viewed in many cultures as an esthetic problem. Consequentially, searching for anti-melanogenic agents from natural sources is very popular worldwide. Previous screening of fermented rice products, obtained from various rice cultivars fermented with different sources of loog-pang (Thai traditional fermentation starter), revealed that the highest ability to reduce the melanin content in B16F10 melanoma cells was from unpolished black rice fermented with a defined starter mixture of microbes isolated from loog-pang E11. The aim of this study was to investigate the mechanism of the fermented unpolished black rice (FUBR) on the inhibition of melanogenesis in B16F10 melanoma cells. The strongest reduction of cellular melanin content was found in the FUBR sap (FUBRS). The melanin reduction activity was consistent with the significant decrease in the intracellular tyrosinase activity. The FUBRS showed no cytotoxic effect to B16F10 melanoma or Hs68 human fibroblast cell lines. It also significantly reduced the transcript and protein expression levels of tyrosinase, tyrosinase-related protein 1 (TYRP-1), TYRP-2, and microphthalmia-associated transcription factor. Furthermore, it induced a significantly increased level of phosphorylated ERK, p38 and Akt signaling pathways, which likely contributed to the negative regulation of melanogenesis. From these results, a model for the mechanism of FUBRS on melanogenesis inhibition was proposed. Moreover, these results strongly suggested that FUBRS possesses anti-melanogenesis activity with high potential for cosmeceutical application as a skin depigmenting agent.

The Protective Effects of Cornus walteri Wanger Leaves against UV Induced Cellular Damage in Human Fibroblast (자외선에 의한 세포손상에 대한 말채나무잎의 보호효과)

  • Park, Hyun-Chul;Jung, Taek Kyu;Yoon, Kyung-Sup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.1
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    • pp.1-7
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    • 2015
  • Cornus walteri Wanger has been used in folk medicine in Korea. Ultraviolet (UV) irradiation has been known as a major cause of photo damage in skin. In the present study, research on how to cure damaged cells by UVB was conducted using an extract of Cornus walteri Wanger leaves (CWE), which was treated with an enzyme. CWE was applied to human dermal fibroblasts (HDFs) affected by UVB. UVB-irradiated HS68 cells showed increased caspase-3 activity, phosphorylation of p53, ${\gamma}H2AX$, cyclobutane pyrimidine dimers (CPDs) formation, and DNA fragmentation compared with non-irradiated cells. However, all these effects were inhibited by treatment with CWE for 12 h after UVB irradiation. Furthermore, CWE has proved not to cause primary skin irritation through the human patch test. Collectively, these results suggest that CWE could be a new potential candidate as photoprotective agent against UVB-induced cellular damage in HDFs.

Antioxidative Activity of Extract of Cornus walteri Wanger Leaves in Human Dermal Fibroblast Irradiated by UVB (자외선이 조사된 섬유아세포에서 말채나무 잎추출물의 항산화작용)

  • Park, Hyun-Chul;Jung, Taek Kyu;Yoon, Kyung-Sup
    • KSBB Journal
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    • v.29 no.6
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    • pp.432-436
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    • 2014
  • We investigated antioxidative activity of the ethanol extracts of leaves of Cornus walteri Wanger (CWE) by treated enzyme in human dermal fibroblast (HDFs) irradiated by UVB. We examined the in vitro chemical and cellular antioxidant activities of CWE in HDFs. We employed scavenging assay for the 1,1-diphenyl-2,5-picrylhydrazyl (DPPH) radicals and cellular antioxidative activity of CWE, and we was investigated in $H_2O_2$-treated or UVB-irradiated HDFs. The CWE effectively scavenged DPPH radicals ($IC_{50}$ $7.03{\pm}0.4{\mu}g/mL$) when compared to the scavenging activities of L-ascorbic acid ($IC_{50}$ $4.69{\pm}0.3{\mu}g/mL$). CWE reduced UVB-induced cellular damage in HS68 cells by MTT assay and inhibited intracellular ROS generation in dose-dependent manner. In addition, CWE also attenuated the elevated levels of 8-isoprostane resulting from UVB-mediated oxidative stress. Collectively, these results suggest that CWE could be a new potential candidate as antioxidant against UVB-induced oxidative stress in HDFs.

Physicochemical Properties and Biological Activities of Collagens with Different Molecular Weights from Alaska Pollack (Theragra chalcogramma) Skin (명태 껍질 유래 콜라겐의 분자량에 따른 이화학적 특성 및 생리활성)

  • Yang, Su-Jin;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1535-1542
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    • 2014
  • This study was conducted to investigate the physicochemical properties and biological activities of collagens with different molecular weights from Alaska pollack (Theragra chalcogramma) skin as well as their efficacies as functional materials. The molecular weights of collagens were between 1~10 kDa (below 1 kDa (AP1), 1~3 kDa (AP2), 3~10 kDa (AP3), and above 10 kDa (AP4). The protein content of AP4 (40.19 g/100 g) was the highest. Collagen contents of AP1, AP2, AP3, and AP4 were 36.43, 32.23, 19.23, and 14.89%, respectively. The free amino acid and essential amino acid contents of AP1 were higher than those of AP2, AP3, and AP4. Fourier transform infrared spectroscopy spectra of collagens with different molecular weights showed wavenumbers representing the regions of amide I, amide II, amide III, and amide A, respectively. The electron-donating ability (29.51%) and SOD-like activity (38.45%) of AP1 were higher than those of AP2, AP3, and AP4. Tyrosinase inhibition activity of AP1 improved with higher treatment concentration. The rate of inhibition of MMP-1 production in HS68 cells exposed to UVB was suppressed by treatment with AP1 (29.78%) and AP2 (26.49%) at 1 mg/mL. Furthermore, there was a strong correlation between DPPH, superoxide dismutase, tyrosinase activity, and MMP-1 inhibition rate of collagens with different molecular weights.