To sum up, the results uncover that the LPS‐TLR4 signalling pathway mediates itch hypersensitivity in mice by regulating the activity of intracellular PI3K and affecting the function of TRPV4 channel, indicating that TLR4‐TRPV4 signalling could be a novel target for the treatment of infection‐related pruritus. Here, TLR4 is linked to Pruritus.