The application discloses a simulation smoke exposure experimental box which comprises a box body and a sealing cover, wherein a smoke inlet is arranged at the bottom part of the box body, a smoke outlet is arranged on the side wall of the top part, an annular groove is arranged at the top part, and liquid is contained in the annular groove; and the sealing cover is covered on the annular groove, an annular vertical wall is arranged thereon, the annular vertical wall is embedded in the annular groove, and the bottom part of the annular vertical wall is immersed in the liquid, thereby not only sealing the box body, but also forming a gas channel between the sealing cover and the annular groove for leading gas in the box body to escape when the pressure in the box body is larger, and further keeping the pressure in the box body constant. Therefore, the simulation smoke exposure experimental box provided by the embodiment of the application can control the gas pressure in the exposure experimental box when carrying out an exposure experiment, avoid an experimental animal in the box from producing barotraumas, lead the smoke inhalation to become the sole wound-causing factor, avoid the bias of the barotraumas and improve the accuracy and the scientificity of exposure experimental data.