Department of Biophysics, Faculty of Medicine, Karadeniz Technical University, Trabzon, Turkey.
Department of Biophysics, Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey.
J Biol Phys. 2022 Sep;48(3):355-368. doi: 10.1007/s10867-022-09612-0. Epub 2022 Aug 10.
There is an increasing interest in the biological and therapeutic effects of fisetin, a natural phenolic compound. Fisetin has affinity on some neuronal targets and may have the potential to modulate neuronal activity. In this study the effects of acute application of fisetin on synchronized events were evaluated electro-physiologically. Besides, interaction of fisetin with closely related channels were investigated in silico. Acute horizontal hippocampal slices were obtained from 32- to 36-day-old C57BL/6 mice. Extracellular field potentials were recorded from CA3 region of the hippocampus. Bath application of 4 aminopyridine (4AP, 100 µM) initiated ictal- and interictal-like synchronized epileptiform discharges in the brain slices. Fifty micromolar fisetin was applied to the recording chamber during the epileptiform activity. The duration and frequencies of both ictal-like and interictal-like activities were calculated from the electrophysiological records. Molecular docking was performed to reveal interaction of fisetin on GABA-A, NMDA, AMPA receptors, and HCN2 channel, which are neuronal structures directly involved in recorded activity. Although fisetin does not affect basal neuronal activity in brain slice, it reduced the duration of ictal-like discharges significantly. Molecular docking results indicated that fisetin has no effect on GABA-A, NMDA, and AMPA receptors. However, fisetin binds to the (5JON) HCN2 channel strongly with the binding energy of -7.66 kcal/mol. Reduction on the duration of 4AP-induced ictal-like discharges can be explained as HCN channels can cause an inhibitory effect via enhancing M-type K + channels which increase K outward currents.
人们对天然酚类化合物漆黄素的生物学和治疗作用越来越感兴趣。漆黄素对一些神经元靶标具有亲和力,可能具有调节神经元活性的潜力。在这项研究中,我们用电生理方法评估了漆黄素对同步事件的急性应用的影响。此外,我们还在计算机上研究了漆黄素与密切相关通道的相互作用。从 32 至 36 天大的 C57BL/6 小鼠中获得急性水平海马切片。从海马 CA3 区记录细胞外场电位。100µM 4-氨基吡啶(4AP)的浴液应用会在脑切片中引发癫痫样和癫痫样样同步癫痫样放电。在癫痫样活动期间,将 50µM 漆黄素应用于记录室。从电生理记录中计算出癫痫样和癫痫样样活动的持续时间和频率。分子对接用于揭示漆黄素与 GABA-A、NMDA、AMPA 受体和 HCN2 通道的相互作用,这些神经元结构直接参与记录的活动。尽管漆黄素不会影响脑切片中的基础神经元活性,但它显著降低了癫痫样放电的持续时间。分子对接结果表明,漆黄素对 GABA-A、NMDA 和 AMPA 受体没有影响。然而,漆黄素与(5JON)HCN2 通道强烈结合,结合能为-7.66 kcal/mol。4AP 诱导的癫痫样放电持续时间的减少可以解释为 HCN 通道通过增强 M 型 K+通道而产生抑制作用,从而增加外向 K 电流。