鼠降钙素基因相关肽放射免疫分析方法的建立及应用

Establishment of radioimmunoassay method for calcitonin gene-related peptide in rats

目的建立检测鼠降钙素基因相关肽(calcitonin gene related peptide-rat,CGRP-rat)的放射免疫分析方法。方法采用重组鼠CGRP-TG皮内多点免疫新西兰兔,获得高效价抗血清,125I标记CGRP,建立放射免疫分析方法,并检测正常及脑缺血再灌注损伤大鼠、小鼠血浆中CGRP浓度。结果标准曲线范围0.1-24.3ng/ml、批内和批间(CV)分别为2.5%及6.6%、灵敏度为0.1ng/ml、回收率104%-107%。本方法测定损伤大鼠、小鼠血浆中CGRP水平较正常对照组显著升高(P<0.01),并与相应正常值之间存在较好相关性(P<0.05)。结论所建立的CGRP-rat放射免疫分析方法可用于心脑血管疾病模型中CGRP含量的测定。

Objective To establish a radioimmunoassay（RIA） method for measurement of calcitonin gene-related peptide（CGRP） in rats.Methods A radioimmunoassay（RIA） method was established using recombinant rat CGRP-thyroglobulin（TG） and 125I-CGRP in New Zealand rabbits to obtain high titer anti serum.Plasma CGRP levels in rats and mice with cerebral ischemia/reperfusion（I/R） injury and controls were measured.Results The standard curve covered 0.1-24.3ng/ml.The intra-and inter-CVs were 2.5% and 6.6%,respectively.The sensitivity was 0.1ng/ml and the recovery rate was 104%-107%.The plasma CGRP levels were significantly higher in rats and mice with cerebral I/R injury than in controls（P〈0.01） and well correlated with their normal ranges（P〈0.05）.Conclusion The RIA method for measuring CGRP in rats we established can be used in measurement of CGRP levels in models of cardio-cerebral diseases.