Immiscible liquid-liquid interfaces have widespread applications in drug delivery, oil-water separation, oil recovery, emulsion and foam stability, etc. Here, a dense viscoelastic interface film is constructed via the self-adsorption of amphiphilic molybdenum disulfide nanosheets at the oil-water interface, which has significant implications for enhanced oil recovery. The microscopic dispersion characteristics of amphiphilic nanosheets in water phase are characterized using Cryo-EM, and the adsorption regularities of amphiphilic nanosheets in horizontal and vertical directions of oil-water interface are also revealed. Besides the mechanical response and rheological properties of interface adsorption film are systematically studied. Experimental results demonstrate that the amphiphilic nanosheet can self-curl to form a single amphiphilic nanosheet with a size of 30 nm in the water phase as the amphiphilic nanosheets concentration is lower than 50 mg/L. When the amphiphilic nanosheets concentration reaches 100 mg/L, obvious water-phase aggregation behaviors happen. The amphiphilic nanosheets can reduce the oil-water interfacial tension from 18.47 mN/m to 0.27 mN/m by forming multilayer adsorption at the oil-water interface. The adsorption ratio of amphiphilic nanosheets from bulk phase to oil-water interface can reach 97.50 %. In addition, the oil-water interface adsorption film exhibits excellent elastic properties and higher strength because the amphiphilic nanosheets can form a planar network structure at the oil-water interface which can be also directly observed in microfluidic pores. The storage modulus and interface shear viscosity of interface adsorption film are 1.65 Pa and 1.38 mN·s/m, respectively. In theory, the required desorption energy of a single amphiphilic nanosheet from oil-water interface to bulk phase is deduced to be 587 KBT. This work is expected to provide amphiphilic molybdenum disulfide nanosheet as an effective candidate for enhanced oil recovery.