扩张型心肌病大鼠心脏左室内皮素受体及其亚型分析

中国临床药理学与治疗学 ›› 2001, Vol. 6 ›› Issue (4): 322-325.

扩张型心肌病大鼠心脏左室内皮素受体及其亚型分析

方五旺, 阎月英, 段宝祥

南京医科大学附属第一医院心血管内科, 南京 210006

收稿日期:2001-07-10 修回日期:2001-08-06 出版日期:2001-08-26 发布日期:2020-11-30
作者简介:方五旺, 男, 医学博士。

Analysis of endothelin receptor and its subtypes in the left ventricle of rats with dilated cardiomyopathy

FANG Wu-Wang, YAN Yue-Ying, DUAN Bao-Xiang

Depatment of Cardiology, Affiliated First Hospital, Nanjing Medical University, Nanjing 210006

Received:2001-07-10 Revised:2001-08-06 Online:2001-08-26 Published:2020-11-30

摘要/Abstract

摘要： 目的观察扩张型心肌病大鼠左室ETR 及其亚型的变化。方法用125 I-ET-1 建立内皮素受体及其亚型的放射配基分析法, 进行饱和结合试验, 对扩张型心肌病大鼠左室心肌ETR 及其亚型的含量进行分析。结果 (1) 放射配基分析法检测ETR 的基本实验条件为:37 ℃ 孵育60 min, 膜蛋白投放量以20～40 μg 为佳;(2) ET-1 及非选择性拮抗剂bosentan、选择性拮抗剂BQ123、BQ788 等均能竞争抑制125 I-ET-1 与内皮素受体的结合, 而去甲肾上腺素则不能抑制;(3) 正常大鼠左室内皮素受体数量为(92.21±34.34) nmol·kg^-1protein, K d 值为(0.04±0.01) nmol, 扩张型心肌病大鼠左心室内皮素受体数量明显减低, K d 值则无明显变化;内皮素受体亚型A 和B 的比例不论在正常还是在心肌病大鼠均接近2:1, 无统计学差异。结论通过放射配基分析法用125 I-ET-1 可以特异地检测内皮素受体及其亚型;扩张型心肌病大鼠左室内皮素受体较正常SD 大鼠明显降低, 但A 和B 两种受体亚型的比例均无明显变化。

关键词: 内皮素受体, 扩张型心肌病, 放射配基分析

Abstract: Aim To observe the changes of endothelin receptors and their subtypes of left ventricules in normal SD rats and dilated cardiomyopathy rats.Methods To establish the best condi tions of the binding experiment, different protein concent rations, incubation temperature and incubating time were tested with 1 25I-ET-1 ligand respectively.With the selected condi tions, saturation binding experiment s were performed to determine the amount of endothelin receptor and its subtypes in normal SD rats and in dilated cardiomyopathy ones.Results (1) The opt imal incubating temperature was 37 ℃.Under this condition, the binding amount of 1 25 I-ET-1 increased rapidly in 0～30 minutes, and reached to saturation point at 60 minutes, and there was alinear correlation between 1 25I-ET-1 binding amount and cell membrane protein concentration. (2) Endo thelin-1, bosentan, BQ123, BQ788 etc.could competit ively suppress the bound of 1 25 I-ET-1 to endothelin receptors.(3) The amount of endothelin receptor in left vent ricle of dilated cardiomy opathy rats was (92.21±34.34) nmol·kg^-1 protein, which was significantly low than that in normal SD ones.There was no change on the ratio of endothelin receptor subty pes A and B.Conclusion 125I-ET-1 can be used to determine the amount of endo thelin receptor and its subty pes in varied tissues specifically.The amount of endothelin receptor in left ventricle of dilated cardiomyopathy rats is down regulated, but the ratio of endothelin receptor subtype A vs B remains to be 2:1.

Key words: endo thelinre, dilated cardiomyopathy, radioligand binding assay

中图分类号:

R542.2

方五旺, 阎月英, 段宝祥. 扩张型心肌病大鼠心脏左室内皮素受体及其亚型分析[J]. 中国临床药理学与治疗学, 2001, 6(4): 322-325.

FANG Wu-Wang, YAN Yue-Ying, DUAN Bao-Xiang. Analysis of endothelin receptor and its subtypes in the left ventricle of rats with dilated cardiomyopathy[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2001, 6(4): 322-325.

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