Hyperglycemia is considered to induce neuronal apoptosis via activating microglia inflammatory responses, thus involving in the development and progression of diabetic encephalopathy and neurodegenerative disorders. Increasing evidences suggest that androgen exerts neuroprotective functions including antiapoptosis, anti-inflammation and antioxidative stress. In this study, we investigate the anti-inflammatory role of dihydrotestosterone (DHT) in high glucose (HG)-induced neuroinflammatory response in BV-2 microglia. Our results revealed that DHT significantly inhibited HG-induced production of nitric oxide and prostaglandin E2 through suppressing the expression of corresponding regulatory enzymes - inducible NO synthase and cyclooxygenase-2. Also, DHT inhibited HG-induced expression of TNF-α and IL-1β. Moreover, DHT suppressed the toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway. Furthermore, when SH-SY5Y neurons were cultured in HG-treated BV-2 microglial supernatant, DHT pretreatment significantly increased neuronal survival, indicating the neuroprotective role of DHT. Collectively, these results suggest that DHT could protect SH-SY5Y neurons from HG-mediated BV-2 microglia inflammatory damage through inhibiting TLR4/NF-κB signaling, suggesting that maintenance of androgen level in brain might have potential benefit in neurodegenerative diseases, especially in diabetes patients combined with cognitive disorders.