<p>This study aimed to explore the associations between various lipid parameters and the risk of gallstone disease (GSD). A multicenter cross-sectional study was conducted to explore the associations between nine lipid parameters and the risk of GSD. Multivariable logistic regression models, adjusted for covariates, were employed to evaluate these associations. Stratified analyses by age and sex were performed to assess population-specific effects. Data from multiple centers were integrated using meta-analysis to generate pooled odds ratios with 95% confidence intervals. Trial Sequential Analysis (TSA) was implemented to determine the sufficiency of the sample size and the robustness of the associations under cumulative evidence conditions. This multicenter study included 580,935 participants, with a GSD prevalence of 7.43%. After adjusting for covariables, the combined multi-center results showed that total cholesterol, high-density and low-density lipoprotein cholesterol, and non-HDL cholesterol were negatively associated with GSD risk. In contrast, the Castelli risk index I and II, atherogenic coefficient, and atherogenic index of plasma were positively associated with the risk of GSD. Subgroup analyses and sensitivity analyses yielded consistent results. Stratified analysis revealed distinct lipid-gallstone associations between people with cholecystectomy and gallbladder stones. TSA revealed that the required amount of information has been achieved, and the results are conclusive. This study suggests significant associations between lipid metabolism and the risk of GSD. There are differences in the relationship between blood lipid indexes and cholecystectomy and gallstones. These findings not only enhance our understanding of the relationship between lipid metabolism and GSD but also provide a novel perspective for risk assessment and prevention strategies.</p>