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Molecular-Pharmacological Analysis of Periodontal Disease : Interaction between LPS and Kinins on Microcirculation of Oral Region

牙周病的分子药理学分析：脂多糖和激肽对口腔微循环的相互作用

基本信息

批准号：
08672142
负责人：
TODOKI Kazuo
金额：
$ 1.22万
依托单位：
Kanagawa Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672142/
关键词：
Optical biosensor Hypoxanthine Ischemia Chemiluminescence Hydrogen peroxide Luminol Oxygen free radicals Plasuma フリーラジカル

项目摘要

The purpose of this study is twofold : 1) to design and develop an optical biosensor that can be used without renewing reagents in the probe for detection of biologically important analytes, and 2) to apply the hypoxanthine affinity optical biosensor to measuring the purine metabolism in an ischemia-reperfusion models. We developed the chemiluminometric bienzyme sensors on the basis of the horseradish peroxidase catalyzed H_2O_2/luminol reaction ; horseradish peroxidase was coimmobilized with xanthine oxidase. Optical biosensor was developed for the batch analysis system of hypoxanthine in the concentration range between 10^<-8> and 10^<-4> M.Optical biosensor probe with photomultiplier tube (PMP) with an integrated preamplifier was investigated to detect the chemiluminescence (which is enhanced by horseradish peroxidase). Horseradish peroxidase and xanthine oxidase were immobilized entrapping sodium alginate gel. The probe based superoxide anion radicals or H_2O_2 sensor achieve detection limits 10^<-8> M.The concentration of hypoxanthine in rabbit serum was also determined in peripheral blood samples (obtained from auricular vein). Increase in hypoxanthine levels was observed in serum obtained immediately after 90 min of auricular artery occlusion. Although there is conflicting evidence as to whether oxygen-derived free radicals play a major role in reperfusion injury, the relatively sustained nature of enhanced oxygen-derived free radical generation during reperfusion has potential clinical implications. With this optical biosensor, it should be possible to measure oxygen-derived free radical generation in a variety of conditions and with multiple interventions. Such studies may further characterize the role of oxygen free radicals in ischemiaミreperfusion injury.

本研究的目的有两个：1）设计和开发一种光学生物传感器，它可以用于检测生物学上重要的分析物，而无需更新探针中的试剂; 2）将次黄嘌呤亲和光学生物传感器应用于缺血-再灌注模型中嘌呤代谢的测量。基于辣根过氧化物酶催化的H_2O_2/鲁米诺反应，将辣根过氧化物酶与黄嘌呤氧化酶共固定，研制了荧光双酶传感器。建立了次黄嘌呤浓度在10 <-8>~ 10 μ <-4>M范围内的光学生物传感器，并将其用于辣根过氧化物酶（HRP）化学发光的检测。采用海藻酸钠凝胶包埋法固定辣根过氧化物酶和黄嘌呤氧化酶。超氧阴离子自由基探针或H_2O_2传感器的检测限均达到10 μ <-8>M。在耳动脉闭塞90分钟后立即获得的血清中观察到次黄嘌呤水平的增加。尽管氧自由基是否在再灌注损伤中起主要作用存在相互矛盾的证据，但再灌注期间氧自由基生成增强的相对持续性质具有潜在的临床意义。有了这种光学生物传感器，它应该是可能的，以测量氧衍生的自由基产生在各种条件下，并与多种干预。这些研究可进一步阐明氧自由基在缺血再灌注损伤中的作用。