摘要
目的 建立大鼠胃壁细胞胃泌素受体放射配基结合实验方法。方法 以12 5I [Leu15] gastrin17 I为标记配基 ,以大鼠胃壁细胞悬液为受体制备 ,反应体系总体积为 2 0 0 μl,37℃恒温水浴振荡孵育 30min ,以 49型玻璃纤维滤膜分离结合和游离配基。结果 胃泌素受体结合符合简单单位点结合模型 ,Scatchard作图求出胃泌素受体结合参数Bmax=4 46 0 4× 10 -12 mol·L-1,Kd=1 2 49× 10 -10 mol·L-1。单点法求出特异结合位点数 70 3 2sites·cell-1。细胞浓度在0 2 5× 10 9～ 2× 10 9cells·L-1范围内 ,与标记配基特异性结合容量成正比。同一样品特异结合测定方法变异系数为7 0 4%。竞争抑制实验证明12 5I Gastin与大鼠胃壁细胞结合特异性良好。
AIM To establish radioligand binding assay of gastrin receptor in rat gastric parietal cell. METHODS Using 125 I labeled [Leu 15 ] gastrin 17 I as the radioligand, rat gastric parietal cells as the receptor preparations. RESULTS In the range of the study, a Scatchard plot of the binding was made by Ligand program with an equilibrium K d of approximately 1 249×10 -10 mol·L -1 and a maximum binding capacity of 4 4604×10 -12 mol·L -1 . The amount of gastrin binding was strongly associated with the cell concentration of rat gastric parietal cell preparations ranged from 0 25×10 9 to 2×10 9 cells·L -1 . The variant coefficent of the binding assay was 7 04% within the same sample. The competitive inhibiting analyses showed that 125 I gastrin was bound to gastric parietal cell preparations with a high specificity. CONCLUSION The radioligand binding assay of gastrin receptor in rat gastric parietal cell preparations meets the criteria for establishing receptor binding assay.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2001年第6期645-647,共3页
Chinese Pharmacological Bulletin
基金
广东省自然科学基金项目
No 980 6 40
