P(1)和P2受体拮抗剂对NG108-15细胞中β,γ-亚甲基ATP和CGS21680诱导的环磷酸腺苷形成的影响。

Effects of P(1) and P2 receptor antagonists on beta, gamma-methyleneATP- and CGS21680-induced cyclic AMP formation in NG108-15 cells.

作者信息

Ohkubo S, Kimura J, Nakanishi H, Matsuoka I

机构信息

Department of Pharmacology, School of Medicine, Fukushima Medical University, Hikarigaoka 1, Fukushima 960-1295, Japan.

出版信息

Br J Pharmacol. 2000 Jan;129(2):291-8. doi: 10.1038/sj.bjp.0703053.

DOI:10.1038/sj.bjp.0703053

PMID:10694235

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1571837/

Abstract

We have previously shown that ATP increased cyclic AMP in NG108-15 cells, which was inhibited by P(1) receptor antagonist methylxanthines. In the present study, we examined the effects of P(1) and P2 receptor antagonists on cyclic AMP formation induced by beta,gamma-methyleneATP (beta,gamma-MeATP) and CGS21680, an A(2A) adenosine receptor agonist, in NG108-15 cells. 2. beta,gamma-MeATP and CGS21680 increased intracellular cyclic AMP with EC(50) values of 8. 0+/-0.98 microM (n=4) and 42+/-7.5 nM (n=4), respectively. 3. Several P(1) receptor antagonists inhibited both beta,gamma-MeATP- and CGS21680-induced cyclic AMP increase with a similar rank order of potency; ZM241385>CGS15943>XAC>DPCPX. However, the pK(i) values of these antagonists for beta,gamma-MeATP were larger than those for CGS21680. 4. Alloxazine, a P(1) receptor antagonist, and several P2 receptor antagonists (PPADS, iPPADS, reactive blue-2) inhibited beta, gamma-MeATP-induced response, while these antagonists little affected CGS21680-induced one. Suramin was effective only for beta, gamma-MeATP-induced response at 1 mM. 5. 2-chloroadenosine (2CADO) and 2-chloroATP (2ClATP) increased cyclic AMP with similar potencies. The effects of these agonists were both inhibited by ZM241385, but only 2ClATP-induced response was inhibited by PPADS. 6. ATP- and beta, gamma-MeATP-induced responses were little affected by alpha, beta-methyleneADP, a 5'-nucleotidase inhibitor. 7. These results clearly demonstrate that ATP-stimulated cyclic AMP formation can be distinguished from the A(2A) receptor agonist-induced one by using the several P(1) and P2 receptor antagonists.

摘要

我们之前已经表明，ATP可增加NG108 - 15细胞中的环磷酸腺苷（cAMP），这一作用被P(1)受体拮抗剂甲基黄嘌呤所抑制。在本研究中，我们检测了P(1)和P2受体拮抗剂对β,γ - 亚甲基ATP（β,γ - MeATP）和CGS21680（一种A(2A)腺苷受体激动剂）在NG108 - 15细胞中诱导的cAMP形成的影响。2. β,γ - MeATP和CGS21680增加细胞内cAMP，其半数有效浓度（EC(50)）值分别为8.0±0.98微摩尔/升（n = 4）和42±7.5纳摩尔/升（n = 4）。3. 几种P(1)受体拮抗剂以相似的效价顺序抑制β,γ - MeATP和CGS21680诱导的cAMP增加；ZM241385 > CGS15943 > XAC > DPCPX。然而，这些拮抗剂对β,γ - MeATP的解离常数负对数（pK(i)）值大于对CGS21680的。4. 嘌呤受体拮抗剂咯嗪和几种P2受体拮抗剂（PPADS、异PPADS、活性蓝 - 2）抑制β,γ - MeATP诱导的反应，而这些拮抗剂对CGS21680诱导的反应影响很小。苏拉明仅在1毫摩尔时对β,γ - MeATP诱导的反应有效。5. 2 - 氯腺苷（2CADO）和2 - 氯ATP（2ClATP）以相似的效价增加cAMP。这些激动剂的作用均被ZM241385抑制，但只有2ClATP诱导的反应被PPADS抑制。6. ATP和β,γ - MeATP诱导的反应几乎不受5'-核苷酸酶抑制剂α,β - 亚甲基ADP的影响。7. 这些结果清楚地表明，通过使用几种P(1)和P2受体拮抗剂，可以区分ATP刺激的cAMP形成和A(2A)受体激动剂诱导的cAMP形成。