Objective To systematically review the efficacy and safety of major ozonated autohemotherapy in the treatment of ischemic stroke.Methods PubMed, EMbase, The Cochrane Library, Web of Science, CBM, WanFang Data, VIP and CNKI were electronically searched to collect randomized controlled trials (RCTs) of the efficacy and safety of major ozonated autohemotherapy in the treatment of ischemic stroke from inception to July 1st, 2020. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies, then, meta-analysis was performed by using RevMan 5.3 software.Results A total of 25 RCTs involving 3 681 patients were included. The results of meta-analysis showed that the major ozonated autohemotherapy combined with conventional therapy in the treatment of patients with ischemic stroke in terms of total effective rate (RR=1.20, 95%CI 1.15 to 1.25, P<0.001), national institutes of health stroke scale (MD=−3.15, 95%CI −4.72 to −1.59, P<0.001), total cholesterol (MD=−1.00, 95%CI −1.48 to −0.53, P<0.001), triglyceride (MD=−0.74, 95%CI −1.04 to −0.43, P<0.001), low density lipoprotein cholesterol (MD=−0.65, 95%CI −1.22 to −0.09, P=0.02), and activity of daily living (MD=11.97, 95%CI 4.48 to 19.47, P=0.002) were superior than the conventional treatment group. There was no significant difference between the two groups in high density lipoprotein cholesterol (MD=0.25, 95%CI −0.46 to 0.96, P=0.49), and the incidence of adverse (OR=3.15, 95%CI 0.93 to 10.63, P=0.06).Conclusion The major ozonated autohemotherapy can significantly improve the prognosis of patients with ischemic stroke, and not increase the evidence of adverse. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify above conclusion.