The invention discloses a python-based application multi-thread operation optimization method. The method comprises the following steps that firstly, a threading module of python is imported; secondly, a thread inherited to the threading module is created, a .run() function of the thread is rewritten, and a long-running operation in an original main program is packaged in the thread again; thirdly, a thread pool used for storing the running thread is created at a main entrance of a program, and the data structure of the thread pool is a list; fourthly, the content is deleted in the place wherethe long-running operation is originally called, the packaged thread is called, and a .start ()function is used for starting a specified thread; fifthly, a wx.CallAfter module is called in the specified thread to return data of a server to the main program. According to the method, multiple program operations can be carried out at the same time, and the program can be effectively prevented from entering a fake halt state.