熊果酸和辛酸联合用药对成年斑马鱼戊四氮诱导的癫痫样行为的影响:临床前研究

Ursolic Acid and Caprylic Acid Cocktail Approach Against Pentylenetetrazole-induced Seizures-like Behavior in Adult Zebrafish: Preclinical Study.

作者信息

Sharma Deepali, Walia Simranjeet, Kaur Simranjit, Kapil Lakshay, Singh Charan, Singh Arti

机构信息

Department of Pharmacology, ISF College of Pharmacy, Moga, affiliated to I. K. Gujral Punjab Technical University, Kapurthala Jalandhar, 144603, India.

Department of Pharmaceutical Sciences, School of Sciences, HNB Garhwal University (A Central University), Chauras Campus, Distt. Tehri Garhwal, Uttarakhand, 246174, India.

出版信息

J Mol Neurosci. 2025 Apr 28;75(2):59. doi: 10.1007/s12031-024-02289-1.

Abstract

Epilepsy is a neurological disorder characterized by recurrent seizures. Pentylenetetrazole (PTZ) is a chemoconvulsant that impairs GABAergic and glutamatergic neurotransmission, promoting excitotoxicity and seizures. Ursolic acid (UA) and caprylic acid (CA) have anti-inflammatory, anti-oxidant, and neuroprotective properties. The objective of the present study was to investigate the combined effect of UA and CA on seizures, neuronal damage, and inflammation induced by PTZ in adult zebrafish. Adult zebrafish (~6-8 months old; 470-530 mg, n=20) were randomly assigned to 10 different groups namely- I) vehicle: 10% DMSO (20µl/kg, i.p), II) Diazepam 1.25 mg/kg per se (DZP, i.p), III) UA 150 mg/kg per se (i.p), IV) CA 60 mg/kg per se (i.p), V) PTZ 170 mg/kg (i.p), VI) DZP 1.25 + PTZ 170 (30 min after DZP 1.25), VII) UA 50 + PTZ 170 (30 min after UA 50), VIII) UA 150 + PTZ 170 (30 min after UA 150), IX) CA 60 + PTZ 170 (30 min after CA 60), and X) UA 50 and CA 60 + PTZ 170 (30 min after UA 50 and CA 60) administration followed by seizure scoring, neurobehavioral (Novel tank test and open field test), biochemical [lipid peroxidase (LPO), acetylcholinesterase (AChEs), superoxide dismutase catalase (SOD), and glutathione-s-transferase (GSH)], molecular (TNF-α, IL-10, Nrf-2 and IL-1β), mitochondrial (complex I, II, IV), cell viability assay (MTT assay) and histopathological analysis. UA of both doses and CA decreased mean seizure score, and mean seizure time. Importantly, combination of UA 50 mg/kg and CA 60 mg/kg attenuated seizure-like behavioural scores, decreased mean seizure time, mean seizure score and reduced the frequency of clonic-like seizures (score 4) in PTZ-treated zebrafish. A combination of UA 50 mg/kg and CA 60 mg/kg also prevented oxidative stress in PTZ-challenged fish by decreasing lipid peroxidation, AChEs activity and increasing catalase, GSH and SOD levels. Additionally, the combination therapy prevented inflammatory response by declining TNF-α and IL-1β levels and raising IL-10 and Nrf-2 levels. Moreover, combination of UA 50 mg/kg and CA 60 mg/kg significantly improved mitochondrial complex I, II and IV activities as well as increase mitochondrial viability in MTT assay. Furthermore, morphology of neuronal cell was prevented in combination of UA 50 mg/kg and CA 60 mg/kg as seen in H & E staining. The protective effect of UA 50 mg/kg and CA 60 mg/kg cocktail is comparable with standard drug Diazepam. Together, we demonstrate that UA 50 mg/kg and CA 60 mg/kg cocktail significantly attenuated PTZ-induced seizure-like behaviours, brain oxidative stress, mitochondrial dysfunction and morphological damage of neuronal cell in zebrafish, suggesting the involvement of its strong anti-inflammatory and antioxidant mechanisms in neuroprotection.

摘要

癫痫是一种以反复发作的癫痫发作为特征的神经系统疾病。戊四氮(PTZ)是一种化学惊厥剂,会损害γ-氨基丁酸能和谷氨酸能神经传递,促进兴奋性毒性和癫痫发作。熊果酸(UA)和辛酸(CA)具有抗炎、抗氧化和神经保护特性。本研究的目的是研究UA和CA联合对成年斑马鱼中PTZ诱导的癫痫发作、神经元损伤和炎症的影响。将成年斑马鱼(约6 - 8个月大;470 - 530毫克,n = 20)随机分为10个不同的组,即:I)溶剂对照组:10%二甲基亚砜(20微升/千克,腹腔注射),II)地西泮1.25毫克/千克单独给药(DZP,腹腔注射),III)UA 150毫克/千克单独给药(腹腔注射),IV)CA 60毫克/千克单独给药(腹腔注射),V)PTZ 170毫克/千克(腹腔注射),VI)DZP 1.25 + PTZ 170(DZP 1.25给药后30分钟),VII)UA 50 + PTZ 170(UA 50给药后30分钟),VIII)UA 150 + PTZ 170(UA 150给药后30分钟),IX)CA 60 + PTZ 170(CA 60给药后30分钟),以及X)UA 50和CA 60 + PTZ 170(UA 50和CA 60给药后30分钟)给药,随后进行癫痫评分、神经行为学(新鱼缸试验和旷场试验)、生化指标[脂质过氧化(LPO)、乙酰胆碱酯酶(AChEs)、超氧化物歧化酶过氧化氢酶(SOD)和谷胱甘肽 - S - 转移酶(GSH)]、分子指标(肿瘤坏死因子 - α、白细胞介素 - 10、核因子E2相关因子2和白细胞介素 - 1β)、线粒体指标(复合体I、II、IV)、细胞活力测定(MTT试验)和组织病理学分析。两种剂量的UA和CA均降低了平均癫痫评分和平均癫痫发作时间。重要的是,50毫克/千克的UA和60毫克/千克的CA联合使用可减轻PTZ处理的斑马鱼的癫痫样行为评分,降低平均癫痫发作时间、平均癫痫评分,并减少阵挛样癫痫发作(评分4)的频率。UA 50毫克/千克和CA 60毫克/千克联合使用还通过降低脂质过氧化、AChEs活性并提高过氧化氢酶、GSH和SOD水平,预防了PTZ刺激的鱼的氧化应激。此外,联合治疗通过降低肿瘤坏死因子 - α和白细胞介素 - 1β水平并提高白细胞介素 - 10和核因子E2相关因子2水平,预防了炎症反应。此外,UA 50毫克/千克和CA 60毫克/千克联合使用显著改善了线粒体复合体I、II和IV的活性,并在MTT试验中提高了线粒体活力。此外,如苏木精和伊红染色所示,UA 50毫克/千克和CA 60毫克/千克联合使用可防止神经元细胞形态改变。50毫克/千克的UA和60毫克/千克的CA联合使用的保护作用与标准药物地西泮相当。总之,我们证明UA 50毫克/千克和CA 60毫克/千克联合使用可显著减轻PTZ诱导的斑马鱼癫痫样行为、脑氧化应激、线粒体功能障碍和神经元细胞形态损伤,表明其强大的抗炎和抗氧化机制参与了神经保护作用。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索