摘要
目的 :建立共同稳定表达大鼠 μ阿片受体及人咪唑啉 1受体的哺乳动物细胞表达系统。 方法 :采用脂质体介导的方法 ,将编码大鼠 μ阿片受体和人咪唑啉 1受体的基因导入CHO细胞中 ,以放射配体 受体结合试验分析受体的表达水平 ,用cAMP积累试验分析表达受体的功能。结果 :在共同稳定表达大鼠 μ阿片受体及人咪唑啉 1受体的CHO细胞中 ,表达的 μ阿片受体对3 H 二丙诺啡 (diprenorphine)的Kd 和Bmax值分别为 (0 .2 4± 0 .0 2 )nmol/L和 (1.83±0 .13)pmol/mg蛋白 ;表达的咪唑啉 1受体对3 H 可乐定的Kd 和Bmax值分别为 (3.72± 0 .2 5 )nmol/L和 (4 3.5 9± 6 .83)fmol/10 6细胞。表达的 μ阿片受体抑制福司科林 (forskolin)刺激下cAMP的积累 ,表达的咪唑啉 1受体抑制吗啡慢性处理、纳洛酮催促引起的cAMP超射。结论 :首次建立了共同稳定表达 μ阿片受体和咪唑啉 1受体的细胞模型。
Objective: To stably co-express rat μ opioid receptor and human imidazoline-1 receptor in mammalian cells. Methods: The cDNAs encoding rat μ opioid receptor and human imidazoline-1 receptor were stably transfected into CHO cells using liposome. Radioligand binding assay and cAMP accumulation assay were used to determine densities and functions of the expressed receptors. Results: Rat μ opioid receptor (rMOR) and human imidazoline-1 receptor (hI 1R) were co-expressed in the CHO cells (CHO-μ/I 1). The K d and B max values were (0.24±0.02)nmol/L and (1.83±0.13) pmol/mg protein for rMOR in 3H-diprenorphine binding assay, and (3.72±0.25) nmol/L and (43.59±6.83) fmol/106 cells for hI 1R in 3H-clonidine binding assay, respectively. The expressed MOR inhibited forskolin-stimulated cAMP accumulation, and the expressed I 1R inhibited naloxone-precipitated cAMP overshooting in chronic morphine-treated cells. Conclusion: The model system of co-expressing MOR and I 1R in CHO cells is first established.
出处
《军事医学科学院院刊》
CSCD
北大核心
2004年第4期329-332,共4页
Bulletin of the Academy of Military Medical Sciences
基金
国家重点基础研究发展计划 ("973"计划 )资助项目 ( 2 0 0 3CB5 15 40 0 )
