Research Article| Volume 32, ISSUE 3, 106971, March 2023

Ro25-6981 alleviates neuronal damage and improves cognitive deficits by attenuating oxidative stress via the Nrf2/ARE pathway in ischemia/reperfusion rats


      • Ro25-6981 treatment improves spatial learning and memory in I/R rats.
      • Ro25-6981 treatment alleviates hippocampal neuronal damage in I/R-induced rats.
      • Ro25-6981 treatment ameliorates I/R-induced oxidative stress by activating the Nrf2-ARE signaling pathway.



      Oxidative stress plays a crucial role in the initiation and progression of cerebral ischemia‒reperfusion injury (CIRI). Therefore, ameliorating oxidative damage is considered to be a beneficial strategy for the treatment of CIRI. NMDAR NR2B subunit antagonists have been reported to be beneficial for synaptic plasticity, neuropathic pain, epilepsy, and cerebral ischemia. However, it remains unclear whether the NR2B subunit antagonist Ro25-6981 has any effect on CIRI.


      In this study, the Morris water maze test and passive avoidance test were used to detect spatial learning and memory. Neuronal loss was measured by Nissl staining. The expression of NSE was assayed by immunohistochemistry. The activities of MDA, 8-OHdG, SOD, GSH-Px, GST and CAT were detected by assay kits. Real-time PCR was used to detect the mRNA levels of hippocampal SOD, GSH-Px and HO-1. Western blotting was used to measure the activation of the Nrf2/ARE pathway by Ro25-6981.


      Ro25-6981 ameliorated cognitive deficits and neuronal damage induced by ischemia‒reperfusion (I/R). Neuronal injury was decreased and the expression of NSE was increased in the CA1 regions of the hippocampus of I/R rats after Ro25-6981 treatment. Moreover, Ro25-6981 alleviated the levels of MDA and 8-OHdG by elevating the activities of SOD, GSH-Px, GST and CAT. Meanwhile, the mRNA levels of SOD, GSH-Px and HO-1 were increased in I/R rats after Ro25-6981 treatment. Furthermore, Ro25-6981 promoted the translocation of Nrf2 to the nucleus, promoting the expression of the Nrf2 downstream genes HO-1 and NQO1.


      The present study indicated that the improvement in the antioxidant properties of Ro25-6981 is mediated by the Nrf2/ARE pathway. This is the first study to demonstrate a favorable effect of Ro25-6981 on cognitive impairment in a CIRI rat model, rendering this NR2B subunit antagonist a promising agent for the treatment or prevention of CIRI.



      CIRI (cerebral ischemia‒reperfusion injury), I/R (ischemia‒reperfusion), CNS (central nervous system), Nrf2 (nuclear factor E2 related factor 2), ARE (antioxidant response element), HO-1 (heme oxygenase-1), NQO1 (NAD(P)H: quinone oxidoreductase), SOD (superoxide dismutase), GSH-Px (glutathione peroxidase), GST (glutathione S-transferase), CAT (catalase), NMDA (N-methyl-D-aspartate), NSE (neuron-specific enolase), MDA (malondialdehyde), 8-OHdG (8-hydroxydeoxyguanosine)
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