Abstract: Due to the strong aggregation effect of fluorescent dyes in aqueous media, the fluorescence quantum yield and stimulated emission efficiency are significantly reduced.To avoid this problem, organic solvents are usually used as the working medium. However, as hazardous chemicals, the introduction of organic solvents brings certain costs and special scene restriction to the application of fluorescent dyes. This article uses sulfobutyl-β-cyclodextrin to construct a water-soluble host system and selects Rhodamine B as the research object, forming a supramolecular covalent complex between the two molecules, analyzing the disassembly effect of Rhodamine B molecules in aqueous media. The experimental results showed that the introduction of sulfobutyl-β-cyclodextrin increased the stimulated emission efficiency of Rhodamine B solution from 11.5% to 20.6%. In addition, the article uses REST technology to construct and enhance sampling molecular dynamics simulations of supramolecular complex system, and analyzes the molecular dynamics mechanism of the regulation of Rhodamine B stimulated emission behavior in aqueous media by sulfobutyl-β-cyclodextrin. The research results of this article can provide important theoretical basis for the subsequent application of fluorescent dyes in aqueous media.