Meat digestion and intestinal flora fermentation characteristics are closely related to human dietary health. The present study investigated the effect of different cooking treatments, including boiling, roasting, microwaving, stir-frying, and deep-frying, on the oxidation of chicken protein as well as its structural and digestion characteristics. The results revealed that deep-fried and roasted chicken exhibited a relatively higher degree of protein oxidation, while that of boiled chicken was the lowest (p < 0.05). Both stir-frying and deep-frying led to a greater conversion of the α-helix structure of chicken protein into a β-sheet structure and resulted in lower protein gastrointestinal digestibility (p < 0.05), whereas roasted chicken exhibited moderate digestibility. Further, the impact of residual undigested chicken protein on the intestinal flora fermentation was assessed. During the fermentation process, roasted chicken generated the highest number of new intestinal flora species (49 species), exhibiting the highest Chao 1 index (356.20) and a relatively low Simpson index (0.88). Its relative abundance of Fusobacterium was the highest (33.33%), while the total production of six short-chain fatty acids was the lowest (50.76 mM). Although stir-fried and deep-fried chicken exhibited lower digestibility, their adverse impact on intestinal flora was not greater than that of roasted chicken. Therefore, roasting is the least recommended method for the daily cooking of chicken. The present work provides practical advice for choosing cooking methods for chicken in daily life, which is useful for human dietary health.