Gorodinsky A, Barg J, Belcheva M M, Levy R, McHale R J, Vogel Z, Coscia C J
Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, MO 63104-1079, USA.
J Neurochem. 1995 Oct;65(4):1481-6. doi: 10.1046/j.1471-4159.1995.65041481.x.
Previously, opioid peptide analogues, beta-endorphin, and synthetic opiates were found to inhibit DNA synthesis in 7-day fetal rat brain cell aggregates via kappa- and mu-opioid receptors. Here dynorphins and other endogenous opioid peptides were investigated for their effect on DNA synthesis in rat and guinea pig brain cell aggregates. At 1 microM, all dynorphins tested and beta-endorphin inhibited [3H]thymidine incorporation into DNA by 20-38% in 7-day rat brain cell aggregates. The putative epsilon-antagonist beta-endorphin (1-27) did not prevent the effect of beta-endorphin, suggesting that the epsilon-receptor is not involved in opioid inhibition of DNA synthesis. The kappa-selective antagonist norbinaltorphimine blocked dynorphin A or B inhibition of DNA synthesis, implicating a kappa-opioid receptor. In dose-dependency studies, dynorphin B was three orders of magnitude more potent than dynorphin A in the attenuation of thymidine incorporation, indicative of the mediation of its action by a discrete kappa-receptor subtype. The IC50 value of 0.1 nM estimated for dynorphin B is in the physiological range for dynorphins in developing brain. In guinea pig brain cell aggregates, the kappa-receptor agonists U50488, U69593, and dynorphin B reduced thymidine incorporation by 40%. When 21-day aggregates were treated with dynorphins, a 33-86% enhancement of thymidine incorporation was observed. Because both 7- and 21-day aggregates correspond to stages in development when glial cell proliferation is prevalent and glia preferentially express kappa-receptors in rat brain, these findings support the hypothesis that dynorphins modulate glial DNA synthesis during brain ontogeny.
此前发现,阿片肽类似物、β-内啡肽和合成阿片类药物可通过κ和μ阿片受体抑制7日龄胎鼠脑细胞聚集体中的DNA合成。在此研究了强啡肽和其他内源性阿片肽对大鼠和豚鼠脑细胞聚集体中DNA合成的影响。在1微摩尔浓度下,所有测试的强啡肽和β-内啡肽均可使7日龄大鼠脑细胞聚集体中[3H]胸腺嘧啶核苷掺入DNA的量减少20%至38%。推定的ε拮抗剂β-内啡肽(1-27)并不能阻止β-内啡肽的作用,这表明ε受体不参与阿片类药物对DNA合成的抑制作用。κ选择性拮抗剂诺宾纳托啡胺可阻断强啡肽A或B对DNA合成的抑制作用,提示存在κ阿片受体。在剂量依赖性研究中,强啡肽B在减弱胸腺嘧啶核苷掺入方面的效力比强啡肽A高三个数量级,表明其作用是由一种离散的κ受体亚型介导的。强啡肽B的IC50值估计为0.1纳摩尔,处于发育中脑内强啡肽的生理范围内。在豚鼠脑细胞聚集体中,κ受体激动剂U50488、U69593和强啡肽B使胸腺嘧啶核苷掺入减少40%。当用强啡肽处理21日龄的聚集体时,观察到胸腺嘧啶核苷掺入增加了33%至86%。由于7日龄和21日龄的聚集体都对应于神经胶质细胞增殖普遍且神经胶质细胞在大鼠脑中优先表达κ受体的发育阶段,这些发现支持了强啡肽在脑个体发育过程中调节神经胶质细胞DNA合成的假说。