This study integrated proteomic and phosphoproteomic analyses to systematically delineate the early-stage (0–6 hours) dynamic molecular landscape of lipopolysaccharide (LPS)-induced polarization of murine RAW264.7 macrophages toward the M1 phenotype. The results revealed that both differentially expressed proteins and phosphorylation sites exhibited a distinct two-stage change pattern during early polarization: proteins in Stage I (adaptive regulation stage) were primarily involved in functional groundwork preparation such as inflammatory priming and cell cycle regulation, whereas the persistently up-regulated proteins in Stage II (delayed response stage) were significantly enriched in pathways including NF-κB signaling and interferon response, driving core anti infection and pro-inflammatory functions. Notably, alterations in protein phosphorylation occurred earlier and more rapidly than changes in protein expression, indicating the leading role of post translational modifications in initiating the polarization process. Kinase prediction analysis further identified key kinase families such as CDK and AKT in regulating early phosphorylation events. These findings not only elucidate the dynamic regulatory network during the early phase of macrophage M1 polarization, but also provide potential molecular targets and a theoretical basis for precisely timed intervention in inflammation-related diseases such as autoimmune disorders, infectious diseases, and inflammatory bone diseases.