HAMI 3379, a CysLT2R antagonist, dose- and time-dependently attenuates brain injury and inhibits microglial inflammation after focal cerebral ischemia in rats

Abstract

Cysteinyl leukotrienes (CysLTs) induce inflammatory responses by activating their receptors, CysLT1R and CysLT2R. We have reported that CysLT2R is involved in neuronal injury, astrocytosis, and microgliosis, and that intracerebroventricular (i.c.v.) injection of the selective CysLT2R antagonist HAMI 3379 protects against acute brain injury after focal cerebral ischemia in rats. In the present study, we clarified features of the protective effect of intraperitoneally-injected HAMI 3379 in rats. We found that HAMI 3379 attenuated the acute brain injury 24 h after middle cerebral artery occlusion (MCAO) with effective doses of 0.1-0.4 mg/kg and a therapeutic window of ∼1h. It attenuated the neurological deficits, and reduced infarct volume, brain edema, and neuronal loss and degeneration 24 and 72h after MCAO. RNA interference with i.c.v. injection of CysLT2R short hairpin RNA (shRNA) attenuated the acute injury as well. Also, HAMI 3379 inhibited release of the cytokines IL-1β, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) into the serum and cerebrospinal fluid 24h after MCAO. Moreover, HAMI 3379 ameliorated the microglial activation and neutrophil accumulation in the ischemic regions, but did not affect astrocyte proliferation 72h after MCAO. In comparison, the CysLT1R antagonist pranlukast did not affect microglial activation and IFN-γ release, but inhibited astrocyte proliferation and reduced serum IL-4. Thus, we conclude that HAMI 3379 has a protective effect on acute and subacute ischemic brain injury, and attenuates microglia-related inflammation. CysLT2R antagonist(s) alone or in combination with CysLT1R antagonists may be a novel class of therapeutic agents in the treatment of ischemic stroke.

Keywords: HAMI 3379; antagonist; cerebral ischemia; cysteinyl leukotriene receptor 2 (CysLT(2)R); inflammation; microglia.