Abstract: OBJECTIVE To investigate the protective effect and mechanism of hyperoside(Hyp) on brain tissue of cerebral ischemia-reperfusion rats. METHODS Rat models of middle cerebral artery occlusion/reperfusion(MCAO/R) were established by bolt-line method and divided into sham operation group, model group and Hyp low, medium and high dose group(30, 60, 120 mg·kg^-1). The rats were evaluated by Zea Longa for neurological function and the brain water content was measured. The volume of cerebral infarction in rats was determined by TTC staining, inflammatory factors were detected by ELISA, pathological morphology of neurons in the hippocampus CA1 was observed by HE staining, apoptosis of brain cells was observed by TUNEL staining, and the expressions of TLR4, COX-2 and apoptosis-related proteins were detected by Western blotting. RESULTS Hyp intervention could significantly improve the Zea Longa score of MCAO/R rats(P<0.05), reduce brain water content and cerebral infarction volume, significantly reduce the expression of inflammatory factors(TNF-α, IL-1β, IL-6, ICAM-1, VCAM-1)(P<0.05 or P<0.01), effectively improve the pathological blood changes and apoptosis of neurons in the hippocampus CA1 region, significantly inhibit the expression of TLR4, COX-2, NF-kB, caspase-3, caspase-9 and Bax protein (P<0.05 or P<0.01), and upregulated the expression of Bcl-2 protein(P<0.05). CONCLUSION The protective effect of Hyp on cerebral ischemia-reperfusion injury is related to anti-inflammatory, anti-apoptotic and inhibition of TLR4/COX-2 signaling pathway.