BACKGROUND: Stroke is the most common serious neurological disorder, and in high-income countries, it is the fourth-leading cause of death, long-term disability, and reduced quality of life among adults. Heart rate variability (HRV) can improve autonomic dysfunction, cognitive impairment, and psychological distress in other patient populations, but its effect in patients with acute ischemic stroke is still unclear. We conducted a protocol for systematic review and meta-analysis to evaluate the efficacy of HRV biofeedback in patients with acute ischemic stroke. METHODS: A computerized literature search will be performed in the following electronic databases from their inceptions to October 2022: PubMed, EMBASE, MEDLINE, Cochrane Central Register of Controlled Clinical Trials, China Knowledge Resource Integrated Database, Wanfang Data Information, and Weipu Database for Chinese Technical Periodicals. The risk of bias in the included articles is assessed according to the Risk of Bias Assessment Tool in Cochrane Handbook of Systematic Reviews. Data are analyzed with the Review Manager Version 5.3 software. RESULTS: This paper will provide high-quality synthesis to assess the efficacy of HRV biofeedback in patients with acute ischemic stroke. CONCLUSION: HRV biofeedback may be a promising intervention for improving autonomic function, cognitive impairment, and psychological distress in patients with acute ischemic stroke.