Calcitriol Ameliorates Brain Injury in the Rat Model of Cerebral Ischemia-Reperfusion Through Nrf2/HO-1 Signalling Axis: An in Silico and in Vivo Study

Abstract

ObjectivesCalcitriol has been revealed to exert neuroprotective effects in ischemic stroke; however, its role and the underlying mechanisms in brain injury induced by ischemia are not well known. The purpose of this study was to determine the neuroprotective effects of calcitriol pretreatment and to assess the possible neuroprotective function of nuclear factor erythroid 2-related factor 2 (Nrf2)/ heme oxygenase-1 (HO-1) signalling pathway against brain ischemia/reperfusion (I/R) injury in the rat models which was followed by a bioinformatics approach. MethodsThe experimental I/R model induction was performed in male Wistar rats for 1 h followed by 23 h reperfusion. Calcitriol was administered intraperitoneally for 7 days prior to stroke. Following ischemia induction 24 h later, neurobehavioral deficits and infarction volume were examined. Oxidative stress was assessed by measurement of malondialdehyde (MDA), nitric oxide (NO) and total antioxidant capacity (TAC). The protein and mRNA expression of HO-1 and Nrf2 were determined by western blot and reverse transcription polymerase chain reaction (RT-PCR), respectively. A molecular docking approach was applied to identify the interaction value of Keap1 with calcitriol. ResultsOur data demonstrated that calcitriol significantly decreased infarction volume and ameliorated neurological deficits in brain I/R. MDA and NO levels were decreased and TAC level was elevated significantly after calcitriol pretreatment. Furthermore, calcitriol upregulated the expression of HO-1 and Nrf2 protein and mRNA in ischemic brain. Molecular modelling demonstrated that calcitriol could interact with the pocket of Keap1 by an appropriate binding energy. ConclusionsThe results indicate that calcitriol protects the brain against I/R injury. This effect may pass through inhibition of oxidative stress and Nrf2/HO-1 pathway activation and this may arise by interaction of Keap1 and calcitriol.