• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1320-1324
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• 2006

The pancreatic and neutrophil elastases are associated with several illnesses including lung and vascular diseases, various cancers, and pancreatitis. The development of a potent and specific inhibitor to the elastases could lead to new therapies. In this study, an agar diffusion method was modified to include a substrate-dye conjugate (Elastin-Congo red) as a substrate of elastase and an indicator of elastase inhibitory activity. The Elastin-Congo red agar plates consisted of 0.1 % Elastin-Congo red and 2.5% agar. The elastase and elastase inhibitors were simultaneously loaded into wells, ultimately resulting in halo formations in which the halo diameter decreased as the concentration of elastase inhibitor increased. The concentration of elastase inhibitor in the samples, therefore, was inversely proportional to the halo diameters. This simplified method provided an excellent correlation with the standard microplate technique, which uses a chromogenic substrate. The concentration of elastase inhibitor obtained from the culture supernatant of a recombinant elastase inhibitor produced by the yeast Pichia pastoris was easily determined. This study has established a simple modified and inexpensive agar diffusion method that is potentially useful for the identification, quantification, and screening of new elastase inhibitors.

• 대한한의학회지
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• 제31권2호
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• pp.137-148
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• 2010

Objective: This study aimed to evaluate the protective effects of Gamipalmi-hwan (GPH) on elastase-induced lung cell injury. Materials and Methods: As an in vitro model of emphysema, the current study was performed to investigate potential activity of GPH in regulating injury responses of A549 human type II cell line mediated by elastase treatment. Results: GPH treatment increased the number of A549 cells which was reduced by elastase digestion. Elastin protein level, which was reduced by elastase treatment, was increased by GPH treatment. Labeling intensity with caspase 3 protein in elastase-treated cells was reduced by GPH treatment. Both Erk1/2 and Cdc2 protein levels, which were decreased by elastase treatment, were increased to a level similar to that of the normal cells. mRNA levels encoding IL-$1{\beta}$ and TNF-$\alpha$ were increased by elastase and then down-regulated by GPH. Conclusion: The present data suggest that A549 cells are subjected to inflammatory damage by elastase and can be recovered by GPH treatment. Further studies examining the protective activity of GPH in elastase-treated lung tissue would be useful for therapeutic strategies of emphysema treatment.

• 한국약용작물학회지
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• 제13권6호
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• pp.213-216
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• 2005

본 연구에서는 비교적 가격이 저렴하고 부작용이 없으면서 효능이 우수한 피부 노화 억제 제품을 개발하고자 먼저 다양한 종류의 약용식물을 대상으로 각종 유기용매 추출물을 제조한 후 이들의 elastase 저해 활성을 측정하여 우수 식물을 선발하고 이들로 부터 elastase 저해제 추출 최적조건을 검토하였다. 64종의 약용식물에 대한 다양한 추출물 가운데 복분자 메탄올 추출물의 elastase 저해 활성이 85%로 가장 높았고 이 저해물질은 메탄올로 $50^{\circ}C$에서 12시간 진탕 시켰을 때 가장 많이 추출 되었다.

• 대한약리학회지
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• 제25권1호
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• pp.109-113
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• 1989

사람 혈액속의 elastase와 관련된 질병에 대한 연구는 다양한 저해제의 개발을 동반해 왔으며, 최근 항생제도 그 관심대상이 되고 있다. 두 단계의 액체 크로마토그래피를 거쳐 얻은 고순도의 elastase에 12종의 항생제를 처리하였다. 세포벽합성 저해제로 알려져 있는 penicillin계와 cephalosporin계 항생제를 각각 3종씩 처리한 결과, methicillin과 cefamandole은 10mM 농도에서 elastase 활성을 50% 이상 저해하였지만, 나머지는 거의 10% 미만이었다. 단백질합성 저해제 중 oxytetracycline의 elastase에 대한 저해효과는 10mM 농도에서 95% 이상으로 매우 탁월하였으며 $(IC_{50}=0.3mM)$, gentamicin도 50% 이상 저해하였으나, 다른 aminoglycoside나 chloramphenicol은 역시 10%미만이었다. 실험해 본 항생제 가운데, oxytetracycline, cefamandole, methicillin, gentamicin 등은 elastase에 대한 강력한 저해제였으며, 그 작용기전은 항생제의 알려진 약리학적 기전과는 다른 차원의 모델임이 분명하였다.

• 대한화장품학회지
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• 제35권1호
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• pp.19-25
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• 2009

피부주름 개선에는 콜라겐(collagen)뿐만 아니라 탄력섬유인 엘라스틴(elastin) 등도 기여하는 것으로 보고되고 있다. 더 나아가 자외선에 의하여 사람의 피부에서 광노화 현상이 나타나며 자외선 조사 후 엘라스타제(elastase)의 활성이 증가하기 때문에 엘라스타제의 활성증가는 자외선에 의한 피부 탄성도의 감소 및 주름 생성의 주요원인으로 생각된다. 따라서 본 연구에서는 피부주름 생성에 영향을 미치는 엘라스타제의 활성을 측정하기 위한 모델을 마련하기 위하여 시판되는 두 가지 엘라스타제, 돼지 췌장 엘라스타제(porcine pancreatic elastase)와 사람 호중구 엘라스타제(human neutrophil elastase)를 사용하였으며 다음 세 가지는 정상 사람 섬유아세포(normal primary human fibroblasts), 쥐의 3T3 섬유아세포주(3T3 mouse fibroblasts), 사람의 CCD-25Sk 섬유아세포주(CCD-25Sk human fibroblasts)로부터 elastase를 준비하여 사용하였다. 준비된 5가지 효소의 농도에 기질의 농도 및 배양시간에 따르는 효소의 활성을 비교 평가하였다. 양성대조군으로 사용한 phosphoramidon이 normal human primary fibroblast elastase와 CCD-25Sk fibroblast elastase의 활성을 유의성 있게 억제한 반면에 porcine pancreatic elastase에는 별다른 영향을 미치지 못하였다. 따라서 주름 개선 후보물질 탐색을 위한 엘라스타제의 선정에는 신중한 고려가 필요할 것으로 사료된다.

• 한국미생물·생명공학회지
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• 제23권5호
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• pp.544-549
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• 1995

The alkaline elastase is an extracellular serine protease of the alkalophilic Bacillus strain Ya-B. To increase the gene copy number and the production level of the alkaline elastase Ya-B, we designed, on the B. subtilis chromosome, a gene amplification of the 10.6 kb repeating unit containing amyE, aleE (alkaline elastase Ya-B gene) and tmrB. The aleE was inserted between amyE and tmrB, and B. subtilis APT119 strain was transformed with this amyE-aleE-tmrB-junction region fragment. As a result, we succeeded in obtaining tunicamycin-resistant (Tm$^{r}$) transformants (Tf-1, Tf-2) in which the designed gene amplification of 10.6 kb occurred in chromosome. The transformants showed high productivity of $\alpha $-amylase and alkaline elastase Ya-B. The copy number of the repeating unit (amyE-aleE-tmrB) was estimated to be 25, but plasmid vector (pUC19) was not integrated. The amplified aleE of chromosome was more stable than that of plasmid in absence of antibiotics.

• Clinical and Experimental Pediatrics
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• 제46권9호
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• pp.903-908
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• 2003

목 적 : 가와사끼병은 어린 소아에서 주로 발생하는 급성 열성 발진성 질환으로 치명적인 관상동맥 합병증을 일으킬 수 있다. 이 질환의 급성기에 관찰되는 호중구 증가와 이 때 분비되어 혈관염 유발에 관여하는 elastase는 가와사끼병의 관상동맥 합병증의 병인에 있어 중요한 역할을 할 것으로 생각된다. 이에 이 질환의 아급성기 합병증인 관상동맥 증대와 초기 elastase 활성도와의 상관관계를 분석하여 관상동맥 합병증의 예측인자로서의 elastase 활성도를 알아보고자 한다. 방 법 : 2001년 11월부터 2002년 1월까지 연세의료원 소아과에 가와사끼병으로 입원하여 치료받은 환아 10명을 대상으로 입원 당일과 발병 4주일 후에 말초 혈액 혈장과 호중구에서 elastase의 활성도를 각각 측정하였고 혈액을 채취한 날 심장 초음파 검사를 같이 시행하여 좌, 우측 관상동맥 근위부의 최대내경을 측정하여 상관관계를 비교 분석하였다. 또한, 가와사끼병에서 elastase 활성도가 유의하게 높은 것을 증명하기 위해 대조군으로 다른 열성 질환 환아 15명의 말초혈액 혈장과 호중구 추출물의 elastase 활성도를 측정하여 비교하였다. 결 과 : 1) 가와사끼병 환아들의 혈장 내 elastase 활성도는 $6.19{\pm}0.74U/mL$로 열성 질환군의 $4.86{\pm}1.17U/mL$에 비해 유의하게 높았으나(P<0.05), 호중구 추출물의 elastase 활성도는 가와사끼병군과 열성 질환군 각각에서 $6.35{\pm}1.70A/{\mu}g$과 $7.78{\pm}4.14A/{\mu}g$으로 양군간 유의한 차이를 보이지 않았다. 2) 가와사끼병 환아에서의 초기 혈장 elastase 활성도와 아급 성기 관상동맥의 증대 정도에는 유의한 관계가 없었다. 결 론 : 가와사끼병에서 다른 열성 질환에 비해 혈장 elastase 활성도가 유의하게 증가되어 있으나 아급성기의 관상동맥 증대와 급성기 elastase의 값에는 유의한 상관관계가 없어 관상동맥 합병증을 예측하는 인자로서의 elastase 역할에 대해서는 더 많은 연구가 필요하다고 본다.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.82-91
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• 2009

Objectives : The purpose of this study is to investigate whitening effects and anti-wrinkle effects of a few 80% ethanol extracted herbal medicines. Methods : In the first study, a few 80% ethanol extracted herbal medicines were screened for their inhibitory activities against the tyrosinase. In the second study, a few 80% ethanol extracted herbal medicines were screened for their inhibitory activities against elastase. Results : 1. We showed 28%, 27% and 19% inhibitions of mushroom tyrosinase at 500 $\mu$g/ml concentration of ASR, AIF and ABR extracts and they were showed higher anti-tyrosinase activity than arbutin's. We also could observe that the decreased mushroom tyrosinase activities in RR, CML, LR, AGR and TH extracts. 2. RR, AF and ABR (final concentrstion 1 mg/ml) were appeared 60%, 98%, 83% of inhibitions of elastase activity, and they were showed higher anti-elastase activity than that of ursolic acid. We also could observe that the decreased elastase activities in AIF, AR, LR and CML extracts. Conclusions : These results suggest that ASR, AIF and ABR extracts contribute to the anti-melanin activities and represent potential sources of whitening agent, and RR, AF and ABR extracts contribute to the anti-elastase activities and represent potential sources of anti-wrinkle agent. These results suggest that some herbal medicines could be strong potential sources of inhibition about anti-aging and whitening effects for the skin.

• 대한본초학회지
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• 제26권1호
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• pp.59-63
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• 2011

Objectives : Elastic fibers are found in the skin, lungs, arteries, veins and other structures. The defects of elastic matrix aggravate hypertension which is associated with alteration in the great arteries, arteries, and arterioles. The elastase inhibitors were undergoing in clinical studies about emphysema and pulmonary hypertension. This study was designed to investigate the effect of Astragali Radix extracts (AR) on elastase activity and anti-oxidative effects. Methods : The elastase inhibitory activity and DPPH (1,1-diphenyl-2-picrylhydrazyl) and NO free radical scavenging activities of AR were measured. Results : The elastase activity was significantly inhibited by AR. The significant DPPH and NO free radical scavenging activities were observed in AR as well. Conclusion : AR showed the anti-elastase effects and anti-oxidative activities in vitro. These results suggest that AR may be a possible drug for the treatment of pulmonary emphysema and pulmonary hypertension.

• The Korean Journal of Physiology and Pharmacology
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• 제2권3호
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• pp.385-393
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• 1998

Human neutrophil elastase (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, was purified by Ultrogel AcA54 gel filtration and CM-Sephadex ion exchange chromatography. HNElastase was inhibited by phenylbutazone in a concentration dependent manner up to 0.4 mM, but as the concentration increased, the inhibitory effect gradually diminished. Binding of phenylbutazone to the human neutrophil elastase caused strong Raman shifts at 200, 440, and 1194 $cm^{-1}$. The peak at 1194 $cm^{-1}$ might be evidence of the presence $of\;-N=N-{\Phi}$ radical. The core area of the elastase, according to the visual molecular model of human neutrophil elastase, was structurally stable. A deeply situated active center was at the core area surrounded by hydrophobic amino acids. Directly neighboring the active site was one positively charged atom and two atoms carrying a negative charge, which enabled the enzyme and the drug to form a strong interaction. Phenylbutazone may form a binding, similar to a key & lock system to the atoms carrying opposite charges near the active site of the enzyme molecule. Furthermore, the hydrophobicity of the surrounding amino acid near the active site seemed to enhance the binding strength of phenylbutazone. Binding of phenylbutazone near the active site may cause masking of the active site, preventing the substrate from approaching the active site and inhibiting elastase activity.