Protective effect of furofuranone against cerebral ischemic stroke via activation of PI3k/Akt/GSK 3β signaling pathway

Purpose: To study the protective effects of furofuranone on oxygen and glucose-deprived damage to brain microvascular endothelial cells ( BMECs) in vitro, and in vivo in cerebral ischemic stroke rat model. Methods: BMECs were isolated from the Sprague Dawley rats and deprived of oxygen and glucose. The effect of 10, 20, 30, 40, 50 and 100 mu M furofuranone on the oxygen/glucose-deprived BMECs was studied using Transwell chamber method. A rat cerebral ischemic stroke model was established using middle cerebral arterial occlusion method. Caspase-3 and other proteins, inflammatory cytokines, and other parameters of the brain tissue were evaluated by enzyme-linked assay (ELISA), polymerase chain reaction (PCR) and Western blot as appropriate. Further studies on the brain tissues was carried out by immunochemical analysis and hematoxylin and eosin staining. Results: Furofuranone decreased caspase 3 levels in a dose-based manner in rat BMECs, and significantly reduced the release of lactate dehydrogenase (LDH) in ischemic stroke rat model (p < 0.05). It also led to marked increases in the levels of p PI3k, p Akt and p GSK3 beta in cerebral ischemic stroke rats. Growth-associated protein-43 (GAP-43) and microtubule-associated protein 2 (MAP-2) levels increased in the cerebral ischemic stroke rat brain tissues, in addition to marked increase in ionized calcium-binding adaptor protein (IBA-1) and glial fibrillary acidic protein (GFAP) (p < 0.05). Furofuranone treatment reduced the population of microtubule-associated protein light chain 3 (MAP1LC3A) and Beclin 1-positive cells, and significantly downregulated L selectin, leptin, monocyte chemotactic protein-1 (MCP- 1) and tumor necrosis factor (TNF)-alpha ( p < 0.05). The release of tissue inhibitor of metalloproteinases 1 (TIMP-1) was enhanced in the cerebral ischemic stroke rats by furofuranone treatment. Conclusion: Furofuranone treatment prevents cerebral ischemic stroke-induced damage in rats via phosphorylation of PI3k, Akt and GSK3 beta proteins, and reduction of inflammatory cytokine levels. Therefore, furofuranone may be useful as chemotherapeutic agent for cerebral ischemic stroke.