An immortalized myocyte cell line, HL-1, expresses a functional delta -opioid receptor.
An immortalized myocyte cell line, HL-1, expresses a functional delta -opioid receptor.
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永生化肌细胞系 HL-1 表达功能性 δ -阿片受体。
DOI：
10.1006/jmcc.2000.1241
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发表时间：
2000
影响因子：
5
通讯作者：
Bolling,SF
中科院分区：
文献类型：
--
作者：
Neilan,CL;Kenyon,E;Kovach,MA;Bowden,K;Claycomb,WC;Traynor,JR;Bolling,SF
本研究的特点阿片受体在永生化心肌细胞系，HL-1。mu（μ）、delta（δ）和kappa（κ）受体的选择性配体对[3 H]布马佐辛的置换显示，只有δ -选择性配体可以完全置换特异性[3 H]布马佐辛结合，表明这些细胞中仅存在δ -受体。用δ -拮抗剂纳曲吲哚进行饱和结合研究，得到[~ 3 H]纳曲吲哚的Bmax为32 fmol/mg蛋白，KD值为0.46nM。从竞争结合试验中获得的HL-1细胞膜中各种δ配体与受体的结合亲和力与使用神经母细胞瘤×神经胶质瘤细胞系NG 108 -15的膜获得的亲和力相似。最后，发现各种δ -激动剂刺激[35 S]GTP γ S的结合，证实心脏δ -受体与G-蛋白偶联。DADLE（D-Ala-D-Leu-enkephalin）在该试验中被发现是最有效的，刺激[35 S]GTP γ S的结合至高于基础水平的27%。上述结果表明，HL-1细胞系含有功能偶联的δ -阿片受体，因此提供了一个体外模型，通过该模型研究阿片对心脏阿片受体的直接作用。
The present study characterizes opioid receptors in an immortalized myocyte cell line, HL-1. Displacement of [3H]bremazocine by selective ligands for the mu (μ), delta (δ), and kappa (κ) receptors revealed that only the δ -selective ligands could fully displace specific [3H]bremazocine binding, indicating the presence of only the δ -receptor in these cells. Saturation binding studies with the δ -antagonist naltrindole afforded a Bmaxof 32 fmols/mg protein and a KDvalue for [3H]naltrindole of 0.46 n M. The binding affinities of variousδ ligands for the receptor in HL-1 cell membranes obtained from competition binding assays were similar to those obtained using membranes from a neuroblastoma×glioma cell line, NG108-15. Finally, various δ -agonists were found to stimulate the binding of [35S]GTP γ S, confirming coupling of the cardiac δ -receptor to G-protein. DADLE (D-Ala-D-Leu-enkephalin) was found to be the most efficacious in this assay, stimulating the binding of [35S]GTP γ S to 27% above basal level. The above results indicate that the HL-1 cell line contains a functionally coupled δ -opioid receptor and therefore provides an in vitro model by which to study the direct effects of opioids on cardiac opioid receptors.