Although most studies have reported that activation of transient receptor potential vanilloid 4 (TRPV4) contributes to bladder pain and overactive bladder with a cardinal symptom of acute or chronic cystitis, its involvement in the protective response against bacterial infection in various cultured cells including urothelial cells has also been reported. In the present study, we investigated the potential benefit of intravesical TRPV4 agonist for painful bladder hypersensitivity produced by a rat model of LPS-induced cystitis and whether its effects modulate the LPS signal for cytokine release and macrophage phenotype change. The increased bladder pain-related behaviors and voiding frequency caused by LPS were suppressed by concurrent injection into the rat bladder of a selective TRPV4 agonist, GSK1016790A. Moreover, the production and secretion of proinflammatory cytokines (e.g., CXCL1, CXCL10), are suppressed by the presence of GSK1016790A. Furthermore, TRPV4 activation switched the LPS-stimulated pro-inflammatory M1-type macrophage to the anti-inflammatory M2-type macrophage. These results suggest that that activation of TRPV4 in bladder regulates the proinflammatory response by LPS, and TRPV4 functions may be a promising future therapeutic target for refractory chronic cystitis.