用组织切片放射配基结合分析法测定几种不同受体的方法学应用研究

An Applied Study of Radioligand Binding Assay on Some Different Receptors

本文主要介绍了组织片放射配基结合分析法（RBA），以及在测量大鼠心肌肾上腺素能受体（Beta－AR）、肾脏血管紧张素II受体（AII－AR）和左心室肌雄激素受体（AR）中的具体方法与应用。通过大量的预实验，摸索出：应用组织片方法，在测定不同组织受体的各自的实验条件与步骤，建立各项适宜的反应条件，包括反应液的配方、PH值；孵育的时间与温度；涮洗的时间和次数等。放射配基结合数据经计算机程序分析，并采用重复非线形回归，表明受体与配基间的结合是简单的单位点结合，即反映受体是单一亲和力的受体。此外，我们观察了耐力训练对性成熟前后大鼠心肌Beta－AR、肾脏AII－R以及左心室肌AR的影响。结果发现耐力训练可以使这些受体有下调趋势，反映了细胞储备增加，能量节省化，系生理适应过程。

We have studied different methods and applications of Radioligand Binding Assay (RBA), determining cardiac Beta - adrenergic receptors (Beta - AR), kidney angiotensin II receptors (AII - R) and left ventricular androgen receptors (AR) of rats. Preliminary experiments have been performed to set up the procedures in preparation of tissues. Also, we have set up the specific conditions in the binding assay applied to measure Beta -AR, AII - R and AR. Specific conditions include incubation temperature,the time to equilibrium of both total and nonspecific binding, buffer PH and composition, etc. Radioligand binding data were analyzed using a computer program that performs iterative nonlinear regression to fit the data to equations describing binding of ligand to a single class of binding sites.In addition, we have investigated effects of endurance training on cardiac Beta-AR, kidney AII - R and left ventricular AR of rats before ad after sexual maturity. Results show that 'Down - regulation' of these receptors is revealed after the endurance training. We suggest that 'Down - regulation' of these receptors may be the internal mechanism of 'Energy Econmization'and physiological adaptation.