Abstract: Introduction: Pain is an unpleasant feeling and it is called as the fifth vital sign. Fistula cannulation in hemodialysis patients is a very stressful and painful process. Nurses can help relieve pain with the use of nonpharmacological pain management therapies, independently. This study conducted aimed to investigate and determine the effect of Valsalva maneuver on pain intensity during cannulation of arteriovenous fistula (AVF) in patients undergoing hemodialysis. Materials and Methods: This study was conducted on 35 hemodialysis patients who were selected by convenience sampling in two Amin Medical Center and Hazrat-e Zahra-e Marziye Hospital in Isfahan. Data collection was performed using an interview questionnaire developed by the researcher, Abbey pain scale, and numerical pain rating. Data analysis was done using the descriptive and analytic tests in SPSS Software Version 20. Results: Paired t-test results showed that the score average of objective and subjective pains was reduced significantly after intervention compared to before that (P< 0.001). Wilcoxon test showed that the objective pain is reduced after intervention (P< 0.001). Wilcoxon test showed that the subjective intensity due to the needle insertion had a significant reduction after intervention (P = 0.001). Conclusion: Performing the Valsalva maneuver before the AVF cannulation reduces pain in patients undergoing hemodialysis.