Ursolic acid along with Caprylic acid Ameliorates Pentylenetetrazole Induced Seizures Like Behavior in Adult Zebrafish

Abstract

Abstract Background: Epilepsy is a neurological disorder characterized by recurrent unprovoked seizures. Despite the existence of more than 20 anti-epileptic drugs there is still a need for new treatments which could not only treat symptoms but also cure epileptogenesis. PTZ is a Chemoconvulsant that impair GABAergic and glutamatergic neurotransmission, promoting excitotoxicity and seizures. Therefore PTZ exposure has been considered a suitable protocol to assess seizure-like behaviours. UA is a triterpenoid compound having anti-inflammatory, anti-oxidant and neuro protective action. CA is one of the component of ketogenic diet and it supresses the inflammation. Objective: The objective of the present study was to investigate the effect of combination therapy of UA and CA on seizures, neuronal damage and inflammation induced by PTZ in adult zebrafish.Materials and method: Zebrafishes were pretreated with Diazepam (1.25 mg/kg), UA(50 and 150 mg/kg), CA (60 mg/kg) and combination of UA (50 mg/kg) and CA (60 mg/kg) followed by PTZ (170 mg/kg) i.p administration. Various neurobehavioral, biochemical parameters like lipid peroxidase (LPO), catalase, superoxide dismutase, glutathione-s-transferase (GSH) and acetylcholinesterase (AChEs), molecular parameters such as TNF-α, IL-10, Nrf-2 and IL-1β and mitochondrial parameters like complex I, II, IV and MTT assay, histopathological study were performed to determine the effect of therapy.Result: UA of both doses and CA decreased mean seizure score, mean seizure time . Importantly, combination of UA 50 mg/kg and CA 60 mg/kg attenuated seizure-like behavioral scores, decreased mean seizure time, mean seizure score and reduced the frequency of clonic-like seizures (score 4). Combination of UA 50 mg/kg and CA 60 mg/kg also prevented oxidative stress in PTZ-challenged fish by decreasing lipid peroxidation, acetyl cholinesterase activity and increasing catalase, glutathione-s-transferase and superoxide dismutase 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 activity as well as increase MTT assay. Furthermore, morphology of neuronal cell was prevented in combination of UA 50 mg/kg and CA 60 mg/kg when seen in histopathology. Similarly as observed in DZP group, combination of UA 50 mg/kg and CA 60 mg/kg affect the overall swimming activity of fish, suggesting different mechanisms of action. Collectively, we show that combination of UA 50 mg/kg and CA 60 mg/kg attenuates PTZ-induced seizure-like behaviours, brain oxidative stress, mitochondrial and morphological damage of neuronal cell in zebrafish, suggesting the involvement of antioxidant mechanisms in neuroprotection.Conclusion: The present study shows that combination of UA 50 mg/kg and CA 60 mg/kg ameliorates the seizures completely and have neuroprotective action via their anti-inflammatory, anti-oxidant properties.