The invention discloses an oil layer protective solution, which is composed of water, a tackifier, a filtrate reducer, an anti-swelling agent, a surfactant, and a bactericide. The protective solution utilizes a synergetic effect of the chemical agents mentioned above so as to realize multiple protective functions on oil layer. The preparation method of the oil layer protective solution comprises the following steps: adding a surfactant and a bactericide into clean water, stirring so as to prepare a basic solution; mixing a tackifier, a filtrate reducer, and an anti-swelling agent according to a ratio so as to make into a uniform solid mixture, then adding the solid mixture into the basic solution with stirring, and stirring to dissolve the mixture so as to form the finished product. The protective solution is used during the process of workover operations such as well killing, well washing, sand washing, pump inspecting, electricity inspecting, electricity to pumping transferring, pumping to electricity transferring, blowing to pumping transferring, blowing to electricity transferring, pump upgrading, and pump depth deepening to protect the oil reservoir so as to prevent the oil reservoir from damages caused by workover fluid.