Department of Biophysics, Institute of Health Sciences, Marmara University, Istanbul, Turkey.
Department of Biochemistry, Institute of Health Sciences, Marmara University, Istanbul, Turkey.
Toxicol Mech Methods. 2023 Feb;33(2):151-160. doi: 10.1080/15376516.2022.2105182. Epub 2022 Jul 31.
Zebrafish () is becoming an increasingly important model in epilepsy research. Pentylenetetrazole (PTZ) is a convulsant agent that induces epileptic seizure-like state in zebrafish and zebrafish embryos and is most commonly used in antiepileptic drug discovery research to evaluate seizure mechanisms. Classical antiepileptic drugs, such as valproic acid (VPA) reduce PTZ-induced epileptiform activities. Opioid system has been suggested to play a role in epileptogenesis. The aim of our study is to determine the effects of morphine in PTZ-induced epilepsy model in zebrafish embryos by evaluating locomotor activity and parameters related to oxidant-antioxidant status, inflammation, and cholinergic system as well as markers of neuronal activity , , and opioid receptors. Zebrafish embryos at 72 hpf were exposed to PTZ (20 mM), VPA (1 mM), and Morphine (MOR) (100 µM). MOR and VPA pretreated groups were treated with either MOR (MOR + PTZ) or VPA (VPA + PTZ) for 20 min before PTZ expoure. Locomotor activity was quantified as total distance moved (mm), average speed (mm/sec) and exploration rate (%) and analyzed using ToxTrac tracking programme. Oxidant-antioxidant system parameters, acetylcholinesterase activity, and sialic acid leves were evaluated using spectrophotometric methods. The expression of , , , and were evaluated by RT-PCR. MOR pretreatment ameliorated PTZ-induced locomotor pattern as evidenced by improved average speed, exploration rate and distance traveled. We report the restoration of inflammatory and oxidant-antioxidant system parameters, , , and opioid receptor as the possible mechanisms involved in the ameliorative effect of MOR against PTZ-induced epileptogenic process in zebrafish embryos.
斑马鱼（）在癫痫研究中正成为一种越来越重要的模型。戊四氮（PTZ）是一种致惊厥剂，可在斑马鱼和斑马鱼胚胎中诱导类似癫痫发作的状态，最常用于抗癫痫药物发现研究中，以评估癫痫发作机制。经典的抗癫痫药物，如丙戊酸（VPA），可降低 PTZ 诱导的癫痫样活动。阿片样物质系统被认为在癫痫发生中起作用。我们的研究目的是通过评估运动活动以及与氧化应激状态、炎症和胆碱能系统相关的参数，以及神经元活动和阿片受体的标志物，来确定吗啡在斑马鱼胚胎 PTZ 诱导的癫痫模型中的作用。在 72 hpf 时，将斑马鱼胚胎暴露于 PTZ（20 mM）、VPA（1 mM）和吗啡（MOR）（100 μM）中。MOR 和 VPA 预处理组在暴露于 PTZ 之前用 MOR（MOR + PTZ）或 VPA（VPA + PTZ）处理 20 分钟。使用 ToxTrac 跟踪程序定量运动活动，作为总移动距离（mm）、平均速度（mm/sec）和探索率（%）进行分析。使用分光光度法评估氧化应激系统参数、乙酰胆碱酯酶活性和唾液酸水平。通过 RT-PCR 评估 、 、 、和 的表达。MOR 预处理改善了 PTZ 诱导的运动模式，表现为平均速度、探索率和移动距离的提高。我们报告了炎症和氧化应激系统参数、、、和阿片受体的恢复，这可能是 MOR 对抗斑马鱼胚胎 PTZ 诱导的致痫过程的改善作用的机制。
