Azilsartan and Ceftriaxone, A Novel Combination Ameliorates Excitotoxicity Mediated Neuroinflammation in In-Vitro, and In-Vivo Models of Cerebral Ischemia

Abstract

Excitotoxicity is an early event of cerebral ischemia, oxidative stress and cytokine storm plays a critical role in dysfunction of Excitatory Amino Acid Transporter-2 (EAAT-2) thereby neuroinflammatory cascade will be activated. Generally, tissue Plasminogen Activators (tPA’s) and anticoagulant therapies are being used as first line treatment options but due to its narrow therapeutic window and anticipatory drawbacks of its administration tPA’s are not much popular in clinical practice. Although, the existing treatment options would not interfere in neuroinflammatory mechanisms rather than restoring the cerebral arterial circulation, in fact EAAT-2 dysfunction during ischemic condition attributes a significant role in neuronal loss. Based on the earlier studies we designed a novel combination for target neuroinflammatory pathways in cerebral ischemia. Angiotensin receptor blocker (ARB) Azilsartan (Azi) and a third generation Cephalosporin Ceftriaxone (Cef) were repurposed to treat the cerebral ischemia inin vitro Oxygen Glucose deprive (OGD) primary astrocytes and N2a neuronal co-culture and in vivo middle cerebral artery occlusion (MCAo) rat model. Novel combination was administered after ischemic reperfusion injury and then we evaluated several parameters such as reactive oxygen species (ROS), apoptosis, oxidative stress markers, cytokine estimation, and RT-PCR. The novel combination ameliorated the neurodegeneration by down regulating the ROS, apoptosis, oxidative stress, glutamate concentration and also enhanced the level of antioxidant enzymes like Superoxide dismutase (SOD), Catalase (CAT) and Reduced Glutathione (GSH). Moreover, EAAT-2 gene expression was remarkably increased with the treatment of novel combination of Azi and Cef than the individual treatment