BackgroundTobacco use is recognized as a major cause of cardiovascular disease, which is associated with endothelial dysfunction. Endothelial function is evaluated using flow-mediated dilation (FMD), which is a noninvasive method. This meta-analysis aimed to investigate the association between smoking exposure and endothelial function evaluated using FMD values.MethodsWe searched the PubMed, Embase, Web of Science, and Cochrane Library databases for cohort studies of smokers or passive smokers that used FMD to assess endothelial function. The primary outcome of the study was the change in the rate of FMD. The risk of bias was evaluated using the Cochrane Collaboration tool and Newcastle–Ottawa Scale. Further, the weighted mean difference was used to analyze the continuous data.ResultsOverall, 14 of 1426 articles were included in this study. The results of these articles indicated that smoking is a major cause of endothelial dysfunction and altered FMD; a pooled effect size of − 3.15 was obtained with a 95% confidence interval of (− 3.84, − 2.46). Notably, pregnancy status, Asian ethnicity, or health status did not affect heterogeneity.ConclusionsWe found that smoking has a significant negative impact on FMD, and measures such as medication or education for smoking cessation may improve endothelial function and reduce the risk of cardiovascular disease.Trial RegistrationThe meta-analysis was registered with PROSPERO on April 5th, 2023 (CRD42023414654).