Under pathological conditions, microglia trigger neurodegeneration by secreting pro-inflammatory molecules, whereas they possess a negative feedback mechanism involving anti-inflammatory cytokines such as IL10. We have previously shown that activation of transient receptor potential vanilloid 4 (TRPV4), a Ca²⁺-permeable cation channels that sense hypo-osmolarity, mechanical stimulus and warm temperature, suppresses LPS-induced microglial activation. Here, we examined whether TRPV4 contributes anti-inflammatory responses using cultured murine microglia and found a selective TRPV4 agonist GSK1016790A enhanced LPS-induced IL10 production, which was fully suppressed by co-application of a selective TRPV4 antagonist GSK2193874 or TRPV4 gene deletion. Furthermore, neutralization of IL10 significantly reversed the inhibitory effects of GSK1016790A on LPS-induced production of pro-inflammatory cytokines. Expression pattern of activation markers implied that GSK1016790A shifted in microglial activation status to an immunoregulatory phenotype. Taken together, these results indicate that microglial TRPV4 plays an important role in promoting the production of LPS-induced IL10, which results in suppressing excessive inflammation in an autocrine or paracrine fashion.