Device and method for measuring two-phase relative permeability curve of unconventional oil reservoir are provided, wherein the device comprises: two-dimensional porous seepage microscopic model; injection components connected to inlet end of the two-dimensional porous seepage microscopic model; confining pressure components arranged outside the two-dimensional porous seepage microscopic model; a camera component arranged on one side of the two-dimensional porous seepage microscopic model; back pressure components connected to outlet end of the two-dimensional porous seepage microscopic model; and outlet pressure measuring and recovery components connected to outlet end of the two-dimensional porous seepage microscopic model. Two-phase relative permeability curve of unconventional oil reservoir can be measured accurately through the device.