• Journal of Periodontal and Implant Science
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• 제29권1호
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• pp.31-40
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• 1999

Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.

• 한국수산과학회지
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• 제44권5호
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• pp.456-463
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• 2011

We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

• 미생물학회지
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• 제40권2호
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• pp.111-114
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• 2004

단삼(Salvia miltiorrhiza) 추출물의 치주질환유발 세균에 대한 생육저지 활성 및 collagenase저해 활성을 측정하였다. 단삼 에탄올 추출물은 치주질환유발 세균에 대해 우수한 생육저지 활성을 나타내었다. 유기용매 분획물중, hexane에서 생육억제능이 가장 우수하였다. 치주질환유발 세균 C. curvus, C. rectus, E. corrodens, F. nucleatum, P. gingivalis, P. intermedia 및 W. succinogenes에 대한 단삼 추출물의 최소생육저해농도(MIC)는 200,50,50,250,150,250및 200 ${\mu}g$/ml로 각각 우수한 생육억제능을 나타내었다. 단삼 유기용매 분획물의 collagenase저해 활성은 동일 농도의 minocycline에 비해 우수한 저해 활성을 나타내었으며, chloroform분획물의 경우 88.2%의 저해 활성을 나타내었다.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.269-274
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• 2002

A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.

• 생명과학회지
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• 제16권2호
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• pp.245-252
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• 2006

피부 트러블을 가진 남,여 40명의 피부에서 분리한 collagenase를 생산하는 균주를 분리, 동정한 결과 Staphylococcus aureus로 판명되었으며 이를 S. aureus JJ-11이라 명명하였다. S. aurells JJ-11 균주의 collagenase의 최적 생산 조건은 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4% (w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$를 함유한 배지 (pH 7.0)에서 $37^{\circ}C$, 200 rpm으로 18시간 동안 배양하는 것이다. 분리 균주가 생산하는 collagenase를 정제하기 위해서 amberlite IRA-900과 sephacryl S-300 HR columns를 이용하였고, 6.66-folds로 정제되었다. S. allreus JJ-11 균주가 생산하는 collagenase를 정제한 결과 분자량은 약 62 kDa이었으며, pH 7.0과 $37^{\circ}C$에서 각각 최대의 활성을 가졌고, pH와 온도에 대한 안정성은 pH 4.0-8.0, $40^{\circ}C$까지 100%의 활성이 있었다. 금속이온에 대해서는 $Fe^{2+},\;Co^{2+},\;Ba^{2+}$ 존재 하에서는 5 mM 농도에서도 활성을 유지하였다. $Sr^{2+},\;Hg^{2+}$에서는 30% 이상이 저해를 받는 것으로 확인되었다. 또한 EDTA와 O-phenanthroline에 의해 65% 이상이 저해되는 일 반적인 collagenase의 특정인 metalloproteinase의 특정을 보였으며, 그리고, 여러 가지 기절에 대해 효소활성을 비교한 결과, insoluble collagen (type I)에 대해 효소 활성이 가장 높았다.

• 생명과학회지
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• 제6권4호
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• pp.241-249
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• 1996

Vivrio mimicus (ATCC 33568)의 최적 배양조건하에서 배양액으로부터 collagenase를 황산암모늄 염석과 DEAE-Sephadex A-50 이온코환크로마토그래피에 의해 분리. 정제하였다. SDS-PAGE 전기영동분석법 및 겔여과법으로 정제된 collagenase의 분자량은 42 kDa 이였다. 기질인 불용성 콜라겐(Type I)에 대한 collagenase의 최적 pH 및 온도는 각각 7.75 및 28$\circ$였다. 금속착물제와 serine protease 저해제는 collagenase의 활성을 저해하였지만 L-cysteine과 histidine은 효소의 활성을 저해하지 않았다. collagenase의 아미노산 조성은 glycine 및 alanine의 아미노산 잔기가 많이 함유되어 있었다. 불용성 (Type I) 콜라겐에 대한 collagenase의 속도상수인 K$_{m}$ 값 및 R$_{cat}$/K$_{m}$값은 각각 2.86 mg/ml 및 972.28 U/mg-protein 이었다.

• 동의생리병리학회지
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• 제25권3호
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• pp.528-532
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• 2011

This study was designed to investigate the collagen metabolism and tyrosinase activity of Meretrix extracts (ME). The effect of ME on type I procollagen production and collagenase activity in human normal fibroblasts HS68 after UVB (312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of ME was measured as well. Type I procollagen production was recovered by ME in UVB damaged HS68 cells. The increased collagenase activity after UVB damage was significantly recovered by ME. The tyrosinase activity and L-DOPA oxidation were significantly reduced as well. However, the effects on tyrosinase activity and L-DOPA oxidation were not powerful enough to be used as whitening agents. ME showed the anti-wrinkle effects and some whitening effects in vitro. These results suggest that ME may be a useful drug as an anti-wrinkle treatments.

• 대한본초학회지
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• 제30권6호
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• pp.1-6
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• 2015

Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

• 혜화의학회지
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• 제21권2호
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• pp.63-72
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• 2013

To develop a new anti-skin aging cosmetics or functional foods by using antioxidative activity and collagenase inhibitor, a potent collagenase or elastase inhibitor was screened from various extracts of medicinal plants and its optimal extraction condition was investigated. And antioxidative activity, antimicrobial activity and inhibitory of effect against collagenase activity were investigated. In the these results, we selected the Sanguisorba (Sanguisorba officinalis L.) that presents a potential biological activities. Sanguisorba which is very rich in triterpenoid saponin and tannins was recently reported its anti-oxidant activities and phytoestogenic activities in vivo test and many clinical studies. The experiments were carried out in vitro to determine anti-oxidant activities of Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, Elastase and collagenase activity assay. It show that the Sanguisorba extracts have the most significant anti-oxidant on free radical scavenging activity assay, and also inhibited significantly activities of elastase, collagenase. Further, Sanguisorba extracts are activated Type I collagen protein expression in CCD-986sk cells. These result suggest that the Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, elastase and collagenase activity assay, Type I collagen protein expression in CCD-986sk cells effected could be developed cosmetic ingredients for anti-aging.

• 대한화장품학회지
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• 제44권3호
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• pp.335-341
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• 2018

본 연구는 버드나무 속 식물의 화장품 소재로써의 활용 가능성을 평가하기 위해 수행하였다. 버드나무 속 식물 중 갯버들, 능수버들 및 키버들 70% 에탄올 추출물의 DPPH radical 소거능 분석을 통한 항산화 활성을 평가한 결과 모든 추출물이 대조군에 비해 DPPH radical 소거능이 유의적으로 증가하였다. 또한 3종의 버드나무 속 식물 추출물 처리는 RAW 264.7 세포의 nitric oxide (NO) 생성을 유의적으로 억제하여 항염 활성이 있는 것으로 확인되었으며, 모두 콜라게나제 저해 활성이 있는 것으로 확인되었다. 그 중 갯버들 추출물이 가장 높은 콜라게나제 저해활성을 나타내어 갯버들 추출물의 용매 분획물에 대한 추가적인 콜라게나제 저해활성을 분석한 결과, 물 및 부탄올 분획물이 가장 높은 콜라게나제 저해활성이 있는 것으로 확인되었다. 결과적으로 버드나무 속 식물 중 갯버들은 높은 콜라게나제 저해활성이 있는 것으로 확인되어 향후 효과적인 기능성 화장품 소재로 활용 가능 할 것으로 사료된다.