The invention discloses a flumazenil liposome injection and a preparation method thereof. The liposome injection is prepared from flumazenil, cholesterol, phosphatidyl ethanolamine, soyasterol, tween 80, trehalose and polyvinylpyrrolidone in specific proportions by weight. The liposome injection disclosed by the invention has favorable preparation stability, the liposome can not be ruptured due to fusion, ice crystal and the like in the freezing process, and the liposome keeps the same favorable entrapment rate after long-term storage. The flumazenil liposome injection disclosed by the invention improves the solubility of flumazenil, improves the quality of the preparation product, reduces the toxic side effect, increases the retention time of the medicament in systemic circulation, improves the bioavailability of the medicament, obviously improves the curative effect, has a simple preparation method and is suitable for industrialized big production.