BACKGROUND: Acute exercise may induce a reduction in pain sensitivity, termed exercise-induced hypoalgesia (EIH). The 45-degree Roman Chair (low back extension) is a dynamic resistance exercise that may be beneficial for low back pain as it allows for direct overload of the spinal flexors and extensors. However, the ability of this exercise to reliably produce EIH has not been examined. PURPOSE: The purposes of this study were to determine the reliability of heat pain threshold (HPT) and pressure pain threshold (PPT) measures, examine local and systemic EIH effects after a 45-degree Roman chair exercise, and determine if EIH was produced reliably between sessions. STUDY DESIGN: Observational Cohort, Crossover. METHODS: Thirty-five healthy participants (mean(SD) age: 21(1.39) years, 16 male) completed two identical sessions at least two days apart. HPT and PPT at the upper trapezius (UT) and low back (LB) were assessed pre/post quiet rest to examine reliability of these measures. HPT and PPT were repeated pre/post one set of the Roman chair exercise until failure to examine reliability of EIH. Intraclass Correlation Coefficients (ICC) examined reliability, and a three-way repeated measures ANOVA compared changes in HPT and PPT among quiet rest and exercise conditions. RESULTS: HPT and PPT demonstrated good (HPT: ICC >0.741) to excellent reliability (PPT: ICC >0.810). Significant EIH as assessed by PPT was demonstrated at both sites (p UT<0.025, LB<0.001) with larger effect sizes at the LB (ηp² >0.413). When assessed with HPT, significant EIH was observed over the LB (p<0.002) but not the UT (p=0.059) EIH can be reliably induced across sessions over the LB (ICC HPT=0.903, PPT=0.815)( HPT=0.903, PPT=0.814) and UT (ICC HPT=0.867, PPT=0.729)( HPT=0.877 and PPT=0.744). CONCLUSION: The 45-degree Roman Chair may reliably induce significant hypoalgesia over the LB assessed with HPT and PPT in healthy participants. LEVEL OF EVIDENCE: 3.