• Title/Summary/Keyword: Collagenase-1

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Retrieval of Porcine Ovarian Follicles by Different Methods

  • Choi, Moon Hwan;Gong, Seung Pyo;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.353-357
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    • 2008
  • A series of studies has been conducted to establish a base infrastructure for an ovarian follicle culture system in the porcine and this study was designed to develop an effective retrieval protocol of preantral follicles. Five different methods using collagenase type I (A) or IV (B, C1, C2 and C3), which employed different treatment durations and/or conditions, were employed and sliced ovarian tissue of prepubertal gilts was provided for the retrieval. A significant increase in total number of follicles retrieved was detected when collagenase IV (methods B or C) was used. In total, more ovarian follicles were retrieved by method B undertaking agitation and method C2 without the agitation than method C1 and C3, while the number of preantral follicles collected was the largest in method B. Neither incubation in 5% $CO_2$ in air atmosphere instead of the agitation nor increased duration of enzymatic treatment up to 120 minutes improved the efficiency of follicle retrieval. There were no differences in the number of follicles retrieved from intact ovaries and from used ovaries for oocyte collection. These results demonstrate the collagenase IV treatment with agitation is effective for retrieving porcine preantral follicles from the ovaries.

Enzymatic Assessment of $2-Hydroxyethyl-{\beta}-undecenate$ Purified from Cumin (Cuminum cymium L.) Seed for Anti-periodontitis (Cumin(Cuminum cymium L.) seed로부터 정제한 $2-hydroxyethyl-{\beta}-undecenate$의 항치주염 효과의 효소학적 평가)

  • Ryu, Il-Hwan;Kang, Eun-Ju;Lee, Kap-Sang;Park, Chung-Soon
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.669-675
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    • 2007
  • The present study was conducted to explore the anti-inflammatory action of $2-hydroxyethyl-{\beta}-undecenate$ (HPS) purified from Cumin (Cuminum cymium L.) seed against periodontitis. From the study in which human leukocyte was employed to detect the inhibiting effects of 5-lipokygenase and cyclooxygenase, enzymes generating carriers of infection like $LTB_4$ and PGs, as well as of collagenase and elastase, organ-destroying enzymes, following conclusions could be drawn: HPS was found to inhibit leukotrien $B_4$ biosynthesis by stimulating more than 97% of human polymorphonuclear leukocyte (PMNL) with addition of $5\;{\times}\;10^{-2}\;M$ when $IC_{50}$ was set at $2\;{\times}\;10^{-4}\;M$. Ninety-two percent of enzyme activation turned out to be inhibited when $5\;{\times}\;10^{-2}\;M$ was added in a test to prove inhibiting effects of HPS against activation of PMNL 5-lipoxygenase from homogeneous humans and purified 5-lipoxygenase on the market. Besides, $IC_{50}$ for enzyme activation was valued at $2.5\;{\times}\;10^{-4}\;M$, while the value of $IC_{50}$ for purified 5-lipoxygenase was $2.3\;{\times}\;10^{-4}\;M$. The $IC_{50}$ values of COX-activated leukocyte and purified collagenase were $5.1\;{\times}\;10^{-4}\;M$ and $2.3\;{\times}\;10^{-4}\;M$, respectively. Moreover, the value of $IC_{50}$ for activation of leukocyte collagenase was $2\;{\times}\;10^{-3}\;M$, whereas that for purified collagenase was $5\;{\times}\;10^{-2}\;M$. In case of leukocyte elastase, addition of $5\;{\times}\;10^{-2}\;M$ inhibited its activation by 66%. In case of purified one, however, activation of enzyme was inhibited by 25% with addition of $5\;{\times}\;10^{-2}\;M$. Furthermore, the $IC_{50}$ value for activation of leukocyte elastase was revealed to be $7.5\;{\times}\;10^{-3}\;M$. From the virulence test with human gingiva cell, it was shown that, on the second day of cultivation, 47.83% of the cell had been activated when HPS was added by $5\;{\times}\;10^{-2}\;M$. Even the addition of HPS by $1\;{\times}\;10^{-2}\;M$ featured 68.53% of cell activation, suggesting relatively strong toxicity of the substance against gingiva cell.

Cellular Changes of Phenotype and Collagenase-1 Expression in Healing Corneal Stromal cells

  • Jung, Jae-Chang
    • Animal cells and systems
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    • v.7 no.3
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    • pp.271-277
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    • 2003
  • Regulation of endoplasmic reticulum(ER) chaperone, ERp29, in traumatized rat spinal cord was investigated. Compared to the control, ERp29 expression was down-regulated at the lesion site 1 d after spinal cord injury. However, ERp29 expression gradually increased from 3 d after the injury and peaked remarkably after 7 d. Two ER chaperones (GRP94 and BiP) showed constantly strong expression levels 1 d after spinal cord injury while the expression levels of the other two (calnexin and PDI) were unchanged. In the case of ERp72, its expression level was increased 1 d after the injury and gradually decreased thereafter. This study suggests that ERp29 expression in the spinal cord after traumatic injury might be associated with the posttraumatic neural survival, playing a role as a molecular chaperone.

Antioxidant and Anti-wrinkling Effects of Extracts from Nelumbo nucifera leaves (하엽(荷葉) 추출물이 항산화 효능 및 피부노화에 미치는 영향)

  • Park, Chan-Ik;Park, Geun-Hye
    • The Korea Journal of Herbology
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    • v.31 no.4
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    • pp.53-60
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    • 2016
  • Objective : The purpose of this study was to investigate anti-aging and antioxidant effects of extracts of Nelumbo nucifera leaves (NN-L) using ethanol on skin .Methods : Each part of leaves(NN-L), flowers(NN-F) and stem(NN-S) was extracted with 70% ethanol. We performed radical scavenging assay(DPPH, ABTS+, Superoxide anion radical), elastase inhibition assay, collagenase inhibition assay. NN-L extracts were tested for cell viability(MTT assay), MMP-1 inhibition and MMP-1 protein expression on CCD-986sk cells (human fibroblast line).Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. We measured its free radical scavenging activity, elastase inhibitory activity and expression of MMP-1 (matrix metalloprotease-1) in human fibroblast cells. Among the parts of Nelumbo nucifera, NN-L showed the highest antioxidant activities and in radical scavenging. DPPH, ABTS+ and Superoxide anion radical scavenging activity of NN-L at concentration of 1,000 μg/mL were 91.43%, 99.31% and 73.7% respectively. In vitro elastase and collagenase inhibition effects of NN-L at concentration of 1,000 μg/mL was 42.8% and 55.3% respectively. The ethanol extract of NN-L showed cell viability of 95.4% in 50 μg/mL concentration. In addition, The results from Western blot assay showed that NN-L decreased the expression of MMP-1 protein in a dose-dependent manner (by up to 35.0% at 50 μM).Conclusion : The findings suggest that the NN-L great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

Study of Cosmeceutical Activities of Hovenia dulcis var. koreana Nakai Extracts (헛개나무 추출물의 화장품 생리활성에 관한 연구)

  • Kim, Sea-Hyun;Jun, Dong-Ha;Jang, Min-Jung;Lee, Jin-Tae;Lee, Chang-Eon;Han, Jin-Gyu;Kim, Jin-Chul;Lee, Do-Hyung
    • Journal of Korean Society of Forest Science
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    • v.99 no.6
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    • pp.836-842
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    • 2010
  • Hovenia dulcis var. koreana Nakai has been reported to liver function improvement effect as functional materials for food and medicine. On these facts, biological activity and safety test were conducted to evaluate biological activities of the fruit petiole and root extracts of H. dulcis as a potential cosmeceutical ingredient. Cosmeceutica activities of different extracts were examined by l.l-diphenyl-2-picrylhydrazyl (DPPH) radical generation, the ABTS+ cation decolorization, tyrosinase activity, collagenase activity and elastase activity compared with the properties of the commercial antioxidant butylated hydroxytoluene (BHT) and L-ascorbic acid (AA). The antioxidant activities HDFW, HDFE, HDRW and HDRE were 83.6%, 39.6%, 85.9% and 74.5% in DPPH assay, 99.5%, 13.7%, 96.4% and 88.6% in ABTS assay. Tyrosinase inhibitiory activities HDFW were 56.0% at 1,000 ppm. Measured the inhibition effect of the H. dulcis about collagenase and elastase where break the peptide bonds in collagen and enzyme from the class of proteases where exists in the dermis. The H. dulcis was inhibition the two kind enzymesm, collagenase activities being on a high scale inhibition, was same concentration. Uses the anti oxidation effect and a anti-wrinkle effect of this resultant H. dulcis and with the functional cosmetics use is thought with the fact that will be possible.

Epidermis Proliferative Effect of the Panax ginseng Ginsenoside $Rb_2$

  • Choi, Seong-Won
    • Archives of Pharmacal Research
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    • v.25 no.1
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    • pp.71-76
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    • 2002
  • Ginseng has been used as a traditional medicine with various therapeutic effects. However, it is still unknown which component of this plant is effective at promoting wound healing. Recently, ginsenoside $Rb_2$ has been reported to improve wound healing. In this study, to investigate the reported wound healing effect of the ginsenoside $Rb_2$, cell morphology and protein factors involved in epidermal formation were evaluated by immunshemical and immunoblotting analysis. $Rb_2$ stimulated epidermal cell proliferation, and the cell showed a 1.5-fold increase in thymidine uptake compared to the control (p<0.05, n=3). Futheremore $Rb_2$, was found to stimulate epidermis formation in a dose-dependent manner in raft culture, and to dose dependently enhance the expressions of protein factors related to cell proliferation, namely, epidermal growth factor and its receptor, fibronectin and its receptor, keratin 5/14, and collagenase 1 (p<0.05, n=3~9). It is believed that ginsenoside $Rb_2$, enhances epidermal cell proliferation by upregulating the expressions of these proliferation-related factors.

Anti-Wrinkle Effects of Enzymatic Oyster Hydrolysate and Its Fractions on Human Fibroblasts (굴 효소 가수분해물과 분자량 분획물의 섬유아세포에 대한 주름 개선 효과)

  • Kim, Hyeon-Ah;Park, Si-Hyang;Lee, Su-Seon;Choi, Yeung Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1645-1652
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    • 2015
  • We investigated the anti-wrinkle efficacy of hydrolysate from oyster protein by Protamex and Neutrase for the purpose of finding materials to assist skin health originating from marine organisms. There were about 7.9% free amino acids in the oyster hydrolysate, and contents of urea, taurine, alanine, and glycine were high. Oyster hydrolysate also showed collagenase inhibitory activity and was not toxic to CCD986sk human fibroblast cells. Yield of the fractions according to the molecular weight of oyster hydrolysate was 40% for less than 1,000 Da and 60.4% for less than 5,000 Da, respectively. Antioxidative effect, procollagen production, and matrix metalloproteinase-1 inhibitory activity were highest in 1,000~3,000 Da fractions. We observed that oyster hydrolysate and its less than 5,000 Da fraction are potential functional compounds for skin health and for improving wrinkles.

Experimental study on the Anti-inflammatory and wound healing effect of Hoichunyanggyuksan and Yongseksa. (回春凉膈散과 龍石散이 抗炎作用에 미치는 影響)

  • Kang, Shung-won;Roh, Seok-seon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.12 no.1
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    • pp.47-78
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    • 1999
  • Hoichunyonggyuksan(HCYGS) and Yongseksan(YSS) are important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\Beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as fo1lows: 1. HCYGS and YSS were not showed the proliferation difference of human fibroblast and monocyte in all concentrations to be experimented and in result, it was concluded that they have no cytotoxicity. 2. YSS inhibited the formation of superoxide to $48\% at the concentration of $0.001\%$ in the mouse monocyte. 3. HCYGS inhibited the formation of superoxide to $13\%$ at the concentration of $0.001\%$ as compared with control in the human monocyte. 4. HCYGS and YSS inhibited the formation of superoxide to $25\%\;and\;35\%$ respectively at the concentration of $0.0001\%\;and\;HCYGS\;showed\;38\%$ inhihitory rate on the formation of superoxide at concentration of $0.001\%$ in the human neutrophil. 5. The concentration of inhibiting the production of prostaglandins($PGE_2$) to $11\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS. 6. The concentration of inhibiting the production of interleukins($IL-l{\Beta}$) to $43\%\;and\;39\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS and YSS. 7. HCYGS and YSS didn't influence on collagen synthesis and total protein in fibroblasts. 8. HCYGS and YSS inhibited the collagenase activity to $22\%\;and\;19\%\;at\;0.1\%,\;45\%\;and\;56\%\;at\;0.2\%,\;57\%\;and\;52\%$ at $0.5\%$ respectively, but YSS exerted the inhibitory effect on collagenase activity to $27\%$ at the lower concentration of $0.01\%$. 9. HCYGS and YSS didn't show the faster recovary than control group hut exerted the consistant effect on the anti-inflammations and the formation of granulation tissue.

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