两种方法测量抗菌肽s-thanatin大鼠体内血药浓度的结果比较  

Comparison of the results of the two methods to determine the concentration of antimicrobial peptide s-thanatin in rat plasma

作　　者：王升兰[1] 李小芳[1] 王西勇[2] 邓学鹏[1] 吴国球[3] 沈子龙[1] 奚涛[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009 [2]东南大学医学院,江苏南京210009 [3]东南大学附属中大医院临床检验中心,江苏南京210009

出　　处：《东南大学学报（医学版）》2011年第3期427-431,共5页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30970809);江苏省自然科学基金资助项目(BK2009279)

摘　　要：目的:分别用间接竞争ELISA法和放射性同位素标记法测定不同时间点抗菌肽s-thanatin(Ts)在动物体内的血药浓度变化,比较两种方法测量结果的差异并分析原因。方法:制备Ts的多克隆抗体,分离纯化多克隆抗体并建立间接竞争ELISA方法的标准曲线。用125I对Ts进行标记,制作同位素标记法的标准曲线。18只SD大鼠随机分为3组,分别按20、10和5 mg.kg-1的剂量尾静脉注射给药(放射性同位素标记法中注射Ts为125I-Ts),于给药后不同时间点眼眶取血,分别用两种方法测定动物体内血药浓度,计算药物半衰期。最后,比较两种方法测量结果的差异,并对其进行分析。结果:间接竞争ELISA法测得Ts在动物体内的半衰期为1.5 h左右,而用放射性同位素标记法测得的半衰期为6 h左右。放射性同位素法测得的生物利用度大约为间接竞争ELISA法的20倍。结论:同位素示踪法测得的药物浓度为动物体内药物代谢物的总量,因此需要借助其它分析仪器才能进行准确的定量试验。Objective： To determine the concentration of antimicrobial peptide s-thanatin（Ts） in rat plasma by competitive indirect ELISA and isotope tracer method,respectively.To compare the differences of the results and find the reasons of the differences.Methods： Prepared the polyclonal antibody of Ts and established the standard curve of competitive indirect ELISA with the purified antibodies.Marked Ts with125I and established the standard curve of isotope tracer method.Eighteen SD rats were divided into 3 groups（n=6 per group）,3 groups rats were given intravenous doses of 5,10 and 20 mg·kg-1 Ts（125I-Ts in isotope tracer method）,respectively.Blood was collected from the retro-orbital plexus in different time after intravenous injection.Determined the concentration of Ts in rats plasma by the two methods and calculated the half-life.Finally compared the differences of the results and found the reasons of the differences.Results： The half-life of Ts in rat plasma was about 1.5 h determined by the competitive indirect ELISA and was about 6 h by isotope tracer method.The AUC which was determined by isotope tracer method was about 20 times as much as that determined by competitive indirect ELISA.Conclusion： The concentration of Ts determined by isotope tracer method is the total metabolites of the drug in rat plasma,so the quantitative test must be done together with other analytical method.

关 键 词：ELISA法 放射性同位素标记 标准曲线 血药浓度 结果比较 

分 类 号：R-332[医药卫生] R965