Newfangled Combination Azilsartan and Ceftriaxone Exhibited Potential Effects in In-Vitro and In-Vivo Models of Cerebral Ischemia: A Comprehensive Pharmacological Investigation.

Abstract

Background: Excitotoxicity is an early event of cerebral ischemia, which is mainly caused by neuroinflammation, oxidative stress, and dysfunction of Excitatory Amino Acid Transporter-2 (EAAT-2). Generally, tissue plasminogen activators (tPA's) and anticoagulant therapies are being used as first-line treatment options for cerebral ischemia, but they only restore the cerebral blood flow (CBF) and fail to attenuate the detrimental events associated with ischemic insult in neurons, resulting in neurodegeneration. Based on the earlier studies, we designed a novel combination for targeting neuroinflammatory and excitotoxicity cascades in cerebral ischemia.

Methods: Angiotensin receptor blocker (ARB) Azilsartan (Azi) and a third-generation cephalosporin Ceftriaxone (Cef) were evaluated in in vitro oxygen glucose-deprived (OGD) primary astrocytes and N2a neuronal co-cultures model of cerebral ischemia. Further, the above combination was also investigated in the middle cerebral artery occlusion (MCAo) rat model of cerebral ischemia. Then, neuro-biochemical estimations and molecular techniques like flow cytometry, ELISA and gene expression studies were performed to elucidate the possible mechanism.

Results: The novel combination ameliorated the neurodegeneration by downregulating the ROS, apoptosis, oxidative stress, and excitotoxicity cascades and also enhanced the level of antioxidant enzymes. Moreover, EAAT-2 gene expression was remarkably increased with the treatment with a novel combination of Azi and Cef than with the individual treatment. The above combination significantly reversed the behavioural dysfunction in ischemic rats, which evidences the beneficial effects.

Conclusion: The repurposing of anti-hypertensive, Azi, and antibiotic Cef combination demonstrated an excellent neuroprotective potential, mediating through ameliorating neuroinflammation, excitotoxicity and oxidative stress in in vitro OGD-induced astrocyte-neuron co-culture as well as cerebral ischemic rat model.