한국과학기술연구원 Korea Institute Of Science and Technology
등록번호
TRKO200200048754
DB 구축일자
2013-04-18
초록▼
Allergy 및 염증반응에 실제 hyaluronidase가 직접적인 관련을 갖는다는 mechanism의 연구가 아직은 덜 되어 있으므로, 우선은 anti-allergic drug 및 anti-inflammatory drug들과 hyaluronidase의 inhibition과의 상관성을 Morgan-Elson법을 변형하여 연구하였다. 이 방법은 in vitro enzyme assay이므로 우선 enzyme의 최적조건의 설정을 살펴, 가장 좋은 optimum condition을 잡아 본 실험에 응용하였다.또 hyaluronidase
Allergy 및 염증반응에 실제 hyaluronidase가 직접적인 관련을 갖는다는 mechanism의 연구가 아직은 덜 되어 있으므로, 우선은 anti-allergic drug 및 anti-inflammatory drug들과 hyaluronidase의 inhibition과의 상관성을 Morgan-Elson법을 변형하여 연구하였다. 이 방법은 in vitro enzyme assay이므로 우선 enzyme의 최적조건의 설정을 살펴, 가장 좋은 optimum condition을 잡아 본 실험에 응용하였다.또 hyaluronidase activation에 필요한 activator를 찾고자 여러금속 1가, 2가, 3가 ion 및 biogenic amine들의 hyaluronidase에 대한 영향을 살펴보았다. 기존에 알려져있는 anti-allergic과 anti-inflammatory drug, antihistamine등 여러종류의 약물의 영향을 살펴 본 결과 anti-allergic drug 및 non-steroidal anti-inflammatory drug들이 hyaluronidase enzyme activity에 dose-dependent하게 inhibition하는 것을 알 수 있었다.실제 이러한 hyaluronidase의 inhibition이 mast cell로 부터의 histamine release에는 어떠한 영향 및 상관성을 나타내는가를 살펴보고자 비만세포(mast cell)로 부터의 histamine release를 Gas Chromatography-Mass Spectrometry의 Negative Chemical Ionization법으로 분석하였다.
Abstract▼
Allergy(hypersensitivity reaction) can be divided into several types such as I (anaphylactic hypersensitivity), II (antibody-dependent cytotoxic hypersensitivity), III (immune-complex mediated hypersensitivity) and IV (cell-mediated delayed type hypersensitivity) according to the initial mechanisms,
Allergy(hypersensitivity reaction) can be divided into several types such as I (anaphylactic hypersensitivity), II (antibody-dependent cytotoxic hypersensitivity), III (immune-complex mediated hypersensitivity) and IV (cell-mediated delayed type hypersensitivity) according to the initial mechanisms, and the treatments used for therapy should be quite different. Among these types of allergy, the type I allergy, which produces detrimental symptoms such as asthma, hay fever and allergic rhinitis, is the most common in man. The pathological mechanism of type I allergy has been explained in terms of the histamine-releasing reaction and the degranulation of mast cells. The degranulation of the mast cells takes place following an immunological stimulus in which the antigen IgE antibody reaction on the membrane of the mast cells predominates and is induced by the influx of the calcium ions into the cells through the calcium channels, and this activates certain enzyme reactions. The mechanism of the degranulation triggered by extracellular calcium ions have not been fully explained yet, and the intracellular series of enzyme reactions has been clarified only to a limited extent.The biochemical mechanism of inflammation is very complicated. However, recent studies suggest that the anti-inflammatory action is attributable to the induction of a group of phospholipase A₂ inhibitory proteins, called lipocortins. Lipocortins are thought to regulate the biosynthesis of potent mediators of inflammation such as prostaglandins, tromboxanes and leukotrienes, by inhibiting the release of their common precursor, arachidonic acid via cyclooxygenase and lipoxygenase.Hyaluronidase is one of the mucopolysaccharide-splitting enzymes, and is related to the permeability of the vascular system and to inflammation. Some enzymes are known to be present in mast cells, and to be probably associated with the release of histamine from the mast cells. In this respect, hyaluronidase usually exists in an inactive form and is activated in vivo by metal ions including calcium ions. Hyaluronic acid, the substrate of this enzyme is also present in substantial quantities in mast cells.So, we considered that hyaluronidase might be a target enzyme of the calcium ions in the degranulation of mast cells, and thus we attempt to investigate the effects of several drugs on the activity of hyaluronidase to apply this approach to simple screening for anti-allergic and anti-inflammatory agents with correlate to hyaluronidase inhibition.Hyaluronidase was markedly inhibited by disodium cromoglycate, a well-known anti-allergic drug and non-steroidal anti-inflammatory drugs(NSAID) such as indomethacin, aspirin, mefenamic acid and ibuprofen etc. No remarkable inhibition was observed in dexamethasone, hydrocortisone, chlorpromazine, mepacrine and diphenhydramine. Since hyaluronidase enzyme assay has revealed a good correlation for the anti-allergic agents and NSAID, it is suggested that the hyaluronidase assay will be a very useful tool for the first screening of anti-allergic agents and NSAID.We are now under investigating the correlationship between hyaluronidase activity and histamine release from rat peritoneal mast cells which is analyzed by negative chemical ionization gas chromatography-mass spectrometry.
목차 Contents
제1장 서 론...23
제2장 실험재료 및 방법...27
제1절 실험재료...27
1. Hyaluronidase 활성 저해법...27
2. Histamine 유리 시험법...28
제2절 실험방법...28
1. Hyaluronidase 활성 저해법...28
2. Histamine 유리시험법...31
2.1 rat 복강내 비만세포의 제조...31
2.2 GC/MS-Negative Chemical Ionization에 의한 Histamine의 분석...32
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