Antibiotic-resistant bacteria can escape from the killing of host immune and settle in the host to form persistent infections. In this study, we investigated the environmental adaptation mechanism of resistant Staphyloccus aureus (S. aureus) to host environment by Data-independent acquisition based quantitative proteomics and functional validation. The detection of growth curve and MIC indicated that ciprofloxacin-resistant S. aureus (Cip-R) showed survival advantage over sensitive strain. Cip-R also exhibited stronger adhesion and invasion ability than sensitive bacteria. Cip-R stimulation resulted in the production of stronger inflammatory factors of the host cells. Proteomics study combined with biochemical validations showed that Cip-R obtains adaptability to host via up-regulation of TCA cycle and down-regulation of ribosome metabolism and protein folding to maintain energy to support their survival. Thus, this study will help us to further explain the growth strategy of resistant bacteria to adapt to the host environment, and provide important information for the development of new antibacterial drugs.