Paudel Yam Nath, Othman Iekhsan, Shaikh Mohd Farooq
Neuropharmacology Research Strength, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Bandar Sunway, Malaysia.
Liquid Chromatography-Mass Spectrometry Platform, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Bandar Sunway, Malaysia.
Front Pharmacol. 2021 Mar 1;11:613009. doi: 10.3389/fphar.2020.613009. eCollection 2020.
Epilepsy is a chronic brain disease afflicting around 70 million global population and is characterized by persisting predisposition to generate epileptic seizures. The precise understanding of the etiopathology of seizure generation is still elusive, however, brain inflammation is considered as a major contributor to epileptogenesis. HMGB1 protein being an initiator and crucial contributor of inflammation is known to contribute significantly to seizure generation via activating its principal receptors namely RAGE and TLR4 reflecting a potential therapeutic target. Herein, we evaluated an anti-seizure and memory ameliorating potential of an anti-HMGB1 monoclonal antibody (mAb) (1, 2.5 and 5 mg/kg, I.P.) in a second hit Pentylenetetrazol (PTZ) (80 mg/kg, I.P.) induced seizure model earlier stimulated with Pilocarpine (400 mg/kg, I.P.) in adult zebrafish. Pre-treatment with anti-HMGB1 mAb dose-dependently lowered the second hit PTZ-induced seizure but does not alter the disease progression. Moreover, anti-HMGB1 mAb also attenuated the second hit Pentylenetetrazol induced memory impairment in adult zebrafish as evidenced by an increased inflection ration at 3 and 24 h trail in T-maze test. Besides, decreased level of GABA and an upregulated Glutamate level was observed in the second hit PTZ induced group, which was modulated by pre-treatment with anti-HMGB1 mAb. Inflammatory responses occurred during the progression of seizures as evidenced by upregulated mRNA expression of HMGB1, TLR4, NF-κB, and TNF-α, in a second hit PTZ group, which was in-turn downregulated upon pre-treatment with anti-HMGB1 mAb reflecting its anti-inflammatory potential. Anti-HMGB1 mAb modulates second hit PTZ induced changes in mRNA expression of CREB-1 and NPY. Our findings indicates anti-HMGB1 mAb attenuates second hit PTZ-induced seizures, ameliorates related memory impairment, and downregulates the seizure induced upregulation of inflammatory markers to possibly protect the zebrafish from the incidence of further seizures through via modulation of neuroinflammatory pathway.
癫痫是一种慢性脑部疾病,全球约有7000万人受其折磨,其特征是持续存在产生癫痫发作的倾向。然而,对癫痫发作产生的病因病理学的确切理解仍然难以捉摸,不过,脑部炎症被认为是癫痫发生的主要因素。HMGB1蛋白作为炎症的引发剂和关键促成因素,已知通过激活其主要受体即RAGE和TLR4对癫痫发作产生有显著贡献,这反映了一个潜在的治疗靶点。在此,我们评估了抗HMGB1单克隆抗体(mAb)(1、2.5和5mg/kg,腹腔注射)在成年斑马鱼中对二次戊四氮(PTZ)(80mg/kg,腹腔注射)诱导的癫痫模型的抗癫痫和改善记忆的潜力,该模型先前用毛果芸香碱(400mg/kg,腹腔注射)刺激过。用抗HMGB1 mAb预处理可剂量依赖性地降低二次PTZ诱导的癫痫发作,但不改变疾病进展。此外,抗HMGB1 mAb还减轻了成年斑马鱼中二次戊四氮诱导的记忆损伤,这在T迷宫试验中3小时和24小时试验时的转折率增加得到证明。此外,在二次PTZ诱导组中观察到GABA水平降低和谷氨酸水平上调,而用抗HMGB1 mAb预处理可对其进行调节。在癫痫发作进展过程中发生了炎症反应,二次PTZ组中HMGB1、TLR4、NF-κB和TNF-α的mRNA表达上调证明了这一点,而用抗HMGB1 mAb预处理后这些表达又反过来下调,反映了其抗炎潜力。抗HMGB1 mAb调节二次PTZ诱导的CREB-1和NPY mRNA表达变化。我们的研究结果表明,抗HMGB1 mAb可减轻二次PTZ诱导的癫痫发作,改善相关记忆损伤,并下调癫痫发作诱导的炎症标志物上调,可能通过调节神经炎症途径保护斑马鱼免受进一步癫痫发作的影响。