Farrelly Lorna A, Savage Niall T P, O'Callaghan Cristina, Toulouse André, Yilmazer-Hanke Deniz M
Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland.
Regul Pept. 2013 Sep 10;186:123-30. doi: 10.1016/j.regpep.2013.08.005. Epub 2013 Aug 28.
Neuropeptide Y (NPY) is a peptide found in the brain and autonomic nervous system, which is associated with anxiety, depression, epilepsy, learning and memory, sleep, obesity and circadian rhythms. NPY has recently gained much attention as an endogenous antiepileptic and antidepressant agent, as drugs with antiepileptic and/or mood-stabilizing properties may exert their action by increasing NPY concentrations, which in turn can reduce anxiety and depression levels, dampen seizures or increase seizure threshold. We have used human neuroblastoma SH-SY5Y cells to investigate the effect of valproate (VPA) and amitriptyline (AMI) on NPY expression at therapeutic plasma concentrations of 0.6mM and 630nM, respectively. In addition, 12-O-tetradecanoylphorbol-13-acetate (TPA) known to differentiate SH-SY5Y cells into a neuronal phenotype and to increase NPY expression through activation of protein kinase C (PKC) was applied as a positive control (16nM). Cell viability after drug treatment was tested with a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. NPY expression was measured using immunofluorescence and quantitative RT-PCR (qRT-PCR). Results from immunocytochemistry have shown NPY levels to be significantly increased following a 72h but not 24h VPA treatment. A further increase in expression was observed with simultaneous VPA and TPA treatment, suggesting that the two agents may increase NPY expression through different mechanisms. The increase in NPY mRNA by VPA and TPA was confirmed with qRT-PCR after 72h. In contrast, AMI had no effect on NPY expression in SH-SY5Y cells. Together, the data point to an elevation of human NPY mRNA and peptide levels by therapeutic concentrations of VPA following chronic treatment. Thus, upregulation of NPY may have an impact in anti-cancer treatment of neuroblastomas with VPA, and antagonizing hypothalamic NPY effects may help to ameliorate VPA-induced weight gain and obesity without interfering with the desired central effects of VPA.
神经肽Y（NPY）是一种存在于大脑和自主神经系统中的肽，它与焦虑、抑郁、癫痫、学习与记忆、睡眠、肥胖以及昼夜节律相关。NPY作为一种内源性抗癫痫和抗抑郁剂最近备受关注，因为具有抗癫痫和/或情绪稳定特性的药物可能通过提高NPY浓度来发挥作用，这反过来又可以降低焦虑和抑郁水平、抑制癫痫发作或提高癫痫阈值。我们使用人神经母细胞瘤SH-SY5Y细胞来研究丙戊酸盐（VPA）和阿米替林（AMI）分别在治疗性血浆浓度0.6mM和630nM时对NPY表达的影响。此外，已知12-O-十四酰佛波醇-13-乙酸酯（TPA）可使SH-SY5Y细胞分化为神经元表型并通过激活蛋白激酶C（PKC）增加NPY表达，将其用作阳性对照（16nM）。药物处理后的细胞活力用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法进行检测。使用免疫荧光和定量逆转录聚合酶链反应（qRT-PCR）测量NPY表达。免疫细胞化学结果显示，VPA处理72小时后NPY水平显著升高，但24小时时未升高。同时用VPA和TPA处理观察到表达进一步增加，表明这两种药物可能通过不同机制增加NPY表达。72小时后用qRT-PCR证实了VPA和TPA使NPY mRNA增加。相比之下，AMI对SH-SY5Y细胞中的NPY表达没有影响。总之，数据表明慢性治疗后治疗浓度的VPA可提高人NPY mRNA和肽水平。因此，NPY的上调可能对用VPA治疗神经母细胞瘤的抗癌治疗有影响，并且拮抗下丘脑NPY的作用可能有助于改善VPA诱导的体重增加和肥胖，而不会干扰VPA所需的中枢作用。
