In this study, three representative pears ('Yali' Pear, 'Huangguan' Pear, and 'Xuehua' Pear) peel/flesh polyphenol extracts were characterized by their antioxidant activity, polyphenol composition, and in vitro cholesterol/cholates binding capacity. 'Yali' Pear polyphenol extracts were selected to further investigate the mechanism of in vitro cholesterol/cholates lowering capacity. Lagergren adsorption kinetic and Freundlich isotherm models confirmed the occurrence of this combination. Turbidity, average particle size, transmission electron microscopy, and zeta potential combined confirmed the existence of some interaction between polyphenols and cholesterol/cholates. Cholesterol/cholates quenched the exogenous fluorescence of polyphenols by static mechanism. The thermodynamic interaction results revealed that the interaction between polyphenols and cholesterol is a spontaneous process, primarily driven by hydrogen bonding and hydrophobic interactions. Overall, this study aimed to investigate the confirmation of the binding removal properties of pear polyphenols on cholesterol/cholates to mitigate the adverse health effects of a high-fat diet.