Aim and objective: To explore the disability, emotional distress and well-being of patients with lumbar spondylolisthesis. Background: Few studies have investigated the correlations between disability, emotional distress and well-being of patients with lumbar spondylolisthesis. Design: This study used a cross-sectional research design. Methods: Participants were 133 patients aged over 50 years who were experiencing lumbar spondylolisthesis. The research instruments included a demographic information questionnaire; the Numeric Rating Scale (NRS); the Charlson Comorbidity Index (CCI); the Chinese versions of the Oswestry Disability Index (ODI), State-Trait Anxiety Inventory-State (STAI-S) and Center for Epidemiological Study-Depression (CES-D); and the Psychological Well-being (PWB) Scale. Emotional distress was measured by the STAI-S and CES-D. Pearson's correlations coefficient, multiple linear regression and a mediating effect model were introduced to explore correlations between the variables and predictors of psychological well-being, and details of the methods are reported in coherence to STROBE criteria. Results: Eighty-six participants (64.6%) had moderate and severe anxiety, and 42 (31.6%) experienced depression. Participants reported medium to high levels of well-being; “satisfaction with interpersonal relationships” was rated the highest and “physical and mental health” the lowest. Disability, depression and anxiety had significant negative correlations with well-being. Depression and anxiety mediated the relationship between disability and well-being. Moreover, depression, family support, educational background and anxiety were predictors of well-being, accounting for 39.1% of the total variance. Conclusions: Disability and emotional distress among patients with lumbar spondylolisthesis had a negative impact on well-being. Anxiety and depression were closely correlated with and substantially influenced well-being. Relevance to clinical practice: Health professionals will enhance the understanding of important factors influencing well-being among patients with lumbar spondylolisthesis. This study suggests the conduct of depression and anxiety evaluations at outpatient clinics and prior to surgery, so that clinicians will be aware of the emotional distress status of patients with lumbar spondylolisthesis and, therefore, enhance their well-being.