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

  • 摘要: 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。

     

    Abstract: 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.

     

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