• Title/Summary/Keyword: Aging degradation

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Auranofin Enhances Sulforaphane-Mediated Apoptosis in Hepatocellular Carcinoma Hep3B Cells through Inactivation of the PI3K/Akt Signaling Pathway

  • Hwangbo, Hyun;Kim, So Young;Lee, Hyesook;Park, Shin-Hyung;Hong, Su Hyun;Park, Cheol;Kim, Gi-Young;Leem, Sun-Hee;Hyun, Jin Won;Cheong, Jaehun;Choi, Yung Hyun
    • Biomolecules & Therapeutics
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    • v.28 no.5
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    • pp.443-455
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    • 2020
  • The thioredoxin (Trx) system plays critical roles in regulating intracellular redox levels and defending organisms against oxidative stress. Recent studies indicated that Trx reductase (TrxR) was overexpressed in various types of human cancer cells indicating that the Trx-TrxR system may be a potential target for anti-cancer drug development. This study investigated the synergistic effect of auranofin, a TrxR-specific inhibitor, on sulforaphane-mediated apoptotic cell death using Hep3B cells. The results showed that sulforaphane significantly enhanced auranofin-induced apoptosis by inhibiting TrxR activity and cell proliferation compared to either single treatment. The synergistic effect of sulforaphane and auranofin on apoptosis was evidenced by an increased annexin-V-positive cells and Sub-G1 cells. The induction of apoptosis by the combined treatment caused the loss of mitochondrial membrane potential (ΔΨm) and upregulation of Bax. In addition, the proteolytic activities of caspases (-3, -8, and -9) and the degradation of poly (ADP-ribose) polymerase, a substrate protein of activated caspase-3, were also higher in the combined treatment. Moreover, combined treatment induced excessive generation of reactive oxygen species (ROS). However, treatment with N-acetyl-L-cysteine, a ROS scavenger, reduced combined treatment-induced ROS production and apoptosis. Thereby, these results deduce that ROS played a pivotal role in apoptosis induced by auranofin and sulforaphane. Furthermore, apoptosis induced by auranofin and sulforaphane was significantly increased through inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway. Taken together, the present study demonstrated that down-regulation of TrxR activity contributed to the synergistic effect of auranofin and sulforaphane on apoptosis through ROS production and inhibition of PI3K/Akt signaling pathway.

Effects of Ethyl Acetate Fraction from Melothria Heterophylla on Antioxidant Activity and Matrix Metalloproteinase-1 Expression in Ultraviolet A-irradiated Human Dermal Fibroblasts (백렴 에틸 아세테이트 층의 항산화 활성과 Matrix Metalloproteinase-1 발현 저해효과)

  • Cho, Young-Ho;;Sim, Gwan-Sub;Lee, Dong-Hwan;Lee, Bum-Chun;Pyo, Heong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.103-109
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    • 2005
  • UV irradiation leads to distinct changes in skin connective tissue, which is degradation of collagen. Many of these alterations in the extracellular matrix are mediated by matrix metalloproteinases. In this study, to develop a new anti-aging agent, we screened the antioxidant activity of solvent fractions from ethanolic extract of Melothria Heterophylla. Among the four solvent fractions tested, the EtOAc fraction exhibited the highest antioxidant activity. It was investigated the inhibitory effect of the EtOAc fraction on the expression and activity of MMP-1 in UVA-irradiated human dermal fibroblasts. The EtOAc fraction inhibited the activity of MMP-1 in a dose dependent manner with the $IC_{50}$ values of $9{\mu}g/mL$. Also, UVA-induced MMP-1 expression was reduced about $90\%$ by $100{\mu}g/mL$ of the EtOAc fraction but MMP-1 mRNA expression was not inhibited. Therefore, we conclude that the EtOAc fraction significantly inhibits MMP-1 expression at the protein level. From these results, we suggest that the EtOAc fraction from M. heterophylla could be used as a new anti-aging agent for the photo-damaged skin.

Change in the Levels of Intracellular Antioxidants during Aging of Articular Chondrocytes and Cartilage (연골세포 및 관절연골의 노화 과정에서 세포내 항산화 인자들의 변화)

  • Kim, Kang Mi;Kim, Yoon Jae;Kim, Jong Min;Sohn, Dong Hyun;Park, Young Chul
    • Journal of Life Science
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    • v.29 no.8
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    • pp.888-894
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    • 2019
  • Cartilage diseases, such as rheumatoid arthritis (RA) and osteoarthritis (OA), are associated with the loss of chondrocytes and degradation of articular cartilage. Recent studies revealed that inflammatory reactive oxygen species (ROS) and age-related oxidative stress can affect chondrocyte activity and cartilage homeostasis. We investigated changes in the levels of intracellular antioxidants during cellular senescence of primary chondrocytes from rat articular cartilages. Cellular senescence was induced by serial subculture (passages 0, 2, 4, and 8) of chondrocytes and measured using specific senescence-associated ${\beta}$-galactosidase ($SA-{\beta}-gal$) staining. ROS production increased significantly in the senescent chondrocytes. In addition, total glutathione (GSSG/GSH) and superoxide dismutase (SOD) levels and heme oxygenase-1 (HO-1) expression increased in senescent chondrocytes induced by serial subculture. Analysis of changes in intracellular antioxidant levels in articular cartilage from rats of different ages (5, 25, 40, and 72 wk) revealed that total glutathione levels were highest after 40 wk and slightly decreased after 72 wk as compared with those after 25 wk. SOD and HO-1 expression levels increased in accordance with age. Based on these results, we conclude that intracellular antioxidants may be associated with cartilage protection against excessive oxidative stress in the process of chondrocyte senescence and age-related cartilage degeneration in an animal model.

Evaluation of skin improvement efficacy of herbal medicine extracts on skin keratinocytes stimulated with fine dust PM10 (미세먼지 PM10으로 손상을 유도한 피부각질형성세포에서 한약재 추출물의 피부 개선 효능 평가)

  • Dong-Hee Kim;Yun Hwan Kang;Bo-Ae Kim
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.4
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    • pp.856-867
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    • 2023
  • Due to the increase in fine dust caused by environmental pollution, oxidative damage and aging of the skin are accelerated. In this study, the antioxidant, hyaluronic acid, filaggrin, MMP-1, and ROS level of selected herbal extracts were evaluated to confirm the protective efficacy of keratinocytes treated PM10. As a result, the antioxidant capacity of 1,1-diphenyl-2-picrylhydrazyl(DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid(ABTS), and FRAP assay increased in a concentration-dependent manner. Keratinocytes the group treated with 300 ㎍/ml of PM10, hyaluronic acid and filaggrin decreased by more than 50%, and increased in the group treated with extracts of Alpinia officinarum, Ulmus macrocarpa, and Ulmus macrocarpa but decreased when the extract was treated, which is evaluated as inhibiting the degradation of collagen and elastin. In addition, in the case of ROS measurement using zebrafish embryos, it was confirmed that the extract was reduced when the extract was treated 25 ㎍/ml, the intensity of fluorescence similar to the negative control was shown, confirming that the production of ROS was significantly reduced. Through this study, the selected oriental medicinal materials, Alpinia officinarum, Ulmus macrocarpa, and Ulmus macrocarpa, protect the skin from fine dust. It is thought that it can be used as an anti-aging product for skin improvement as a material that can be improved or improved.

Behaviour of the soil residues of the herbicide quinclorac in the micro-ecosystem (pot) (Micro-ecosystem(pot)중 제초제 quinclorac 토양잔류물의 행적)

  • Ahn, Ki-Chang;Kyung, Kee-Sung;Lee, Jae-Koo
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.96-106
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    • 1998
  • Rice plants were grown for 42 days in the specially made micro-ecosystem(pot) containing two different soils treated with fresh and 60-day-aged residues of [$^{14}C$]quinclorac, respectively, to elucidate the behaviour of the herbicide quinclorac residues in the soils. Amounts of $^{14}CO_{2}$ evolved from two soils treated with different residues with and without vegetation were all less than 2.2% of the total $^{14}C$, indicating that there was little microbial degradation of quinclorac in soil. $^{14}C$-Radioactivity absorbed and translocated into rice plants from soil A and B containing fresh quinclorac residues was 8.4 and 24.2%, respectively, of the originally applied $^{14}C$, while 5.5 and 17.7%, in aged residue soils. These results indicate that larger amounts of $^{14}C$ were absorbed by rice plants from soil B with less organic matter and clay than soil A, and the uptake of [$^{14}C$]quinclorac and its degradation products decreased with aging in soil. After 42 days of rice growing, 84.5 and 61.8% of the $^{14}C$ applied freshly to soil A and B, respectively, remained in soil, whereas, in the case of aged soils, 86.3 and 67.7% of the $^{14}C$ applied did. Meanwhile, without vegetation, more than 98.3% of the $^{14}C$ applied, in both fresh and aged residues, remained in soil, suggesting that quinclorac was relatively persistent chemically and microbiologically. Most of the non-extractable soil-bound residues of [$^{14}C$]quinclorac were incorporated into the organic matter and largely distributed in the fulvic acid portion.

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Changes of SDS-PAGE Pattern and Allergenicity of BSA and BGG in Beef Extract Treated with Heat and High Pressure (물리적 처리에 따른 우육추출물중의 BSA와 BGG단백질의 SDS-PAGE패턴 및 항원성의 변화)

  • Han, Gi-Dong;Fan, Jiang Ping;Suzuki, Atsushi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.5
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    • pp.594-599
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    • 2006
  • In our previous report, we indicated that not only BSA but also BGG played an important role in the allergenicity of beef. In this study, the effect of heat or high-pressure treatments to beef extract on the SDS-PAGE patterns was examined. The antigenicity of each treated samples was also investigated by Western blots assay with the sera of BGG-positive beef allergic patients. The BGG band and its antigenicity slightly disappeared but not generally in $100^{\circ}C$ group, indicating $100^{\circ}C$ treatment is not sufficient to totally eliminate the antigenicity of beef allergens. Compared with BGG band, BSA band significantly disappeared in SDS-PAGE with $100^{\circ}C$ treatment, indicating BSA is more heat- sensitive than BGG. When the beef extract was heated at $120^{\circ}C$, not only BSA but also BGG bands was largely disappeared in both SDS-PAGE and Western blots. High pressure (HP) treatment even at 600 MPa did not affect SDS-PAGE and Western blots pattern of BSA. On the contrary, BGG treated with HP showed visible changes in SDS-PAGE. 600 MPa treatment significantly reduced the antigencity. Interestingly, these behaviors of BGG were not found in the same experiments with pure BGG treated with HP. From these results, it was speculated that some kinds of proteolytic enzymes in beef extracts were involved in the BGG molecular degradation by HP treatment. The aging experiments of beef extracts treated with HP supported this hypothesis. Further studies are needed to clarify the function and working mechanism of enzymes associated with BGG degradation in beef extracts by HP treatment.

The Effect of Glycolic Acid on Human Dermal Fibroblasts: Increased Collagen Synthesis and Inhibition of MMP-2/9

  • Park, Ki-Sook;Kim, Soo-Kyoum;Lim, Sae-Hwan;Kim, Yun-Young;Park, Young-Ju;Lee, Seung-Soo;Lee, Su-Hvun;Chang, Tae-Hyun;Son, Youna-Sook
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.519-523
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    • 2003
  • Alpha hydroxy acid (AHA) includes a group of organic acids found in natural foods such as sugarcane (glycolic acid), milk (lactic acid), apples (malic acid) and oranges (citric acid). Earlier studies demonstrated the effect of AHAs on the skin by diminishing the adhesiveness of the corneal layer and increasing the viable epidermal thickness. Recent data suggest that AHAs have some effects on the dermal component of skin and even affect the aging process of the skin. A previous study revealed increased collagen production by treatment with glycolic acid among AHAs in vitro. However, the mechanism of the regulation of collagen production by glycolic acid was unclear. In present study, we tried to demonstrate the effect of glycolic acid on human dermal fibroblasts and to unveil the mechanism of regulation of collagen production by glycolic acid in human dermal fibroblasts: proliferation of fibroblasts and collagen synthesis and degradation by collagenases in fibroblasts. Our results suggested that glycolic acid had no effect on proliferation and cytotoxicity of adult human dermal fibroblasts. However, glycolic acid not only induced the increase of the collagen synthesis in human dermal fibroblasts at lower concentration than 0.1 % but also inhibited MMP-2 activity of human dermal fibroblast in the range between 0.01 and 0.4% and MMP-9 activity of human dermal fibroblast in the range between 0.06 and 0.09%. In summary, our results suggest that glycolic acid may increase wrinkle reduction partially by both increase in collagen synthesis and decrease in collagen degradation.

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Study on Stabilization of Retinaldehyde using Drug-in-Cyclodextrinin-Liposome (DCL) for Skin Wrinkle Improvement (레틴알 안정화를 위한 사이클로덱스트린-리포좀에 관한 연구)

  • Ha, Ji Hoon;Choi, Hyeong;Hong, In Ki;Han, Sang-Kuen;Bin, Bum Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.1
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    • pp.77-85
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    • 2022
  • Retinaldehyde (RA), vitamin A derivative, is an intermediate between retinol and retinoic acid and has an excellent wrinkle improving effect. In this study, Drug-in-cyclodextrin-in-liposome (DCL) was used to enhance the stability and skin penetration of RA. The complex of RA and hydroxypropyl-beta-cyclodextrin (HP-β-CD) was prepared by the freeze-drying method, and the presence or absence of inclusion of retinal was confirmed by UV-Vis spectrometer, FT-IR and SEM images. RA was captured in HP-β-CD about 95.6% on 1 : 15 (w/w). The retinal-HP-β-CD complex was encapsulated in liposomes using a homomixer and microfluidizer, with an average particle size of 215 ± 4.2 nm and a zeta potential of -31.2 ± 0.5 mv. In the evaluation of the degradation stability of RA, degradation rate of RA-HP-β-CD-liposomes in water was 1.8% higher than RA-liposome (5.8%), RA-HP-β-CD complex (9.7%) and RA alone (37.6%). RA cream (0.05% RA) including RA-HP-β-CD-liposomes was prepared for clinical test with wrinkle-improving efficacy and skin dermis denseness evaluated for 2 or 4 weeks. RA cream showed a significant wrinkle improving effect without skin irritation. In conclusion, it was confirmed that the double stabilization technology using the DCL system contribu tes to the effect of improving skin wrinkles by increasing the stabilization of retinal.

Effect of Quercetin in the UV-Irradiated Human Keratinocyte HaCaT Cells and A Model of Its Binding To p38 MAPK

  • Jnawali, Hum Nath;Lee, Eunjung;Shin, Areum;Park, Young Guen;Kim, Yangmee
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2787-2790
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    • 2014
  • Quercetin is a major dietary flavonoid found in onions, apples, tea, and red wine, and potentially has beneficial effects on disease prevention. We carried out this study to investigate the effect of quercetin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in human keratinocyte HaCaT cells and to further understand the mechanisms of its action. The anti-inflammatory activity of quercetin was investigated and quercetin significantly suppressed the NO production in LPS-stimulated RAW264.7 mouse macrophages. Post treatment of quercetin decreased UV irradiation-induced phosphorylation of JNK, p38 MAPK, and ERK by 91%, 21%, and 17%, respectively. MMP-1 is mainly responsible for the degradation of dermal collagen during the aging process of human skin and quercetin suppressed the UVB-induced MMP-1 by 94%. Binding studies revealed that quercetin binds to p38 with high binding affinity ($1.85{\times}10^6M^{-1}$). The binding model showed that the 4'-hydroxy groups of the B-ring of quercetin participated in hydrogen bonding interactions with the side chains of Lys53, Glu71, and Asp168 and the 5-hydroxy group of the A-ring formed a hydrogen bond with the backbone amide of Met109. The major finding of this study shows that quercetin inhibits phosphorylation of JNK, p38 MAPK, and ERK pathway leading to the prevention of MMP-1 expression in human keratinocyte HaCaT cells. Therefore, our findings suggested the potentials of quercetin as a skin anti-photoaging agent.

Case Study on Network of Manpower-training related to Long-term care insurance system - Focus on Education management about Long-term care-giver of Yong Do Gu in Busan city - (장기요양보험제도에 따른 인력양성의 네트워크 사례연구 - 부산시 영도구 요양보호사 교육운영 사례 -)

  • Nam, Hee Eun;Lim, Chang Ho;Ryu, Hwang Gun;Bae, Sung Kwon;Kim, Sang Hee;Kim, Sun Hee;Lee, Jae Hee;Kim, Hwang Eun
    • The Korean Journal of Health Service Management
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    • v.2 no.1
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    • pp.125-136
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    • 2008
  • Government came to enforce long-term care insurance system in preparation for the rapid aging society. Whether this system is successful or not depends on the professionalism of long-term care-givers who are professional population in charge of care service. Currently in the early stage of enforcement, such problems as a race cutting fee resulted from numerical increase of educational facilities, insolvent operation, degradation of education level resulted from unprofessional instructor, are pointed out. As a mean of manpower-training on long-term care insurance system, this study is to research public-private-university network model of the Academy of Continuing Education attached to Ko Sin University which is the case of Yong Do Gu Busan city. Networking between the vision and development strategy of Yong Do Gu on continuing education city, education system on community manpower-training supported by Ko Sin University, and service field of welfare for the elderly can not only contribute to the professionalism of long-term care-givers but also play an ideal role in manpower-utilization and arrangement of community. Through this networking, high quality of education level and circumstance, using the existing infra, manpower-training and utilization for continuing education of Yong Do Gu can be accomplished. Additionally, the connection with facilities related with welfare for the elderly can contribute to professionalism and accountability of manpower-networking.

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