SD大鼠二胺氧化酶活力测定方法的研究

Studies on Determination of Enzymatic Active for Diamine Oxidase in Sprague-dawley rats

目的建立SD大鼠二胺氧化酶活力测定方法。方法采用分光光度法,酶反应体系包括底物戊二胺,催化酶二胺氧化酶和辣根过氧化物酶,邻联二茴香胺和磷酸盐缓冲盐溶液,反应体系pH为7.2,反应温度为37℃,检测波长460nm。结果二胺氧化酶在2.5～40mg/ml浓度范围内均呈良好的线性关系,r=0.9983(n=5)。血清二胺氧化酶平均回收率为97.74%,RSD为1.51%;小肠二胺氧化酶的平均回收率为97.49%,RSD为1.01%。稀释实验测定值与稀释度呈线性相关,相关系数为0.9982。结论本方法简便、灵敏、稳定、可靠,作为特征酶可应用于肠缺血再灌注MODS疾病模型的研究。

Objective To establish a method for determination of enzymatic active for diamine oxidase in Sprague-dawley rats. Methods A Colorimetric method was used, the enzymatic reaction system consisted of Cadaverine dihydrochloride for substrate, Diamine oxidase and Horseradish Peroxidase for kinase , O-Dianisidine dihydrochloride and phosphate buffer solution （ adjust to a pH of 7.2） and the detection wavelength was 460nm. Results Diamine oxidase had a good linearity r = 0. 9983 in the range of 2. 5 - 40 mg , ml-1 ,its average recovery of samples was 97.74% （ RSD = 1.51%, n = 6） and 97. 49% （ RSD = 1.01%, n = 6）. The experimental correlation coefficient of dilution is found to be 0. 9982. Conclusion The method was simple, sensitive, lasting and reliable for simultaneous determination of diamine oxidase in serum and intestine, and was used for the studies on disease model of mutiple organ dyfunction syndrome induced by intestinal ischemia-reperfusion in rats as characteristic enzyme.