Background: The application of high PEEP remains to be a controversial issue when it comes to ICU patients underwent ventilation. There are studies supporting the usage of high PEEP in patients with ARDS, while for those without ARDS, the conclusion is of great ambiguity. We performed this systematic review and meta-analysis to compare the effects of high and low level of PEEP on ICU patients without ARDS.Methods: We searched public databases (including PubMed, EMBASE, Cochrane Library and Clinicaltrial.gov) to find eligible randomized controlled trials (RCTs). The primary outcomes included in this meta-analysis were in-hospital mortality, 28-day mortality and the duration of ventilation, ICU stay, and hospital stay. We used the Cochrane risk of bias assessment tool to evaluate risk of bias. Trial Sequential Analysis (TSA) was conducted. Results: We included 2307 patients from 24 trials using high and low PEEP. Although no significant difference was found between high and low PEEP applications in in-hospital mortality (risk ratio[RR] 0.98, 95% confidence interval[CI] [0.81, 1.19], P=0.87), 28-day mortality (RR 0.68, 95% CI [0.33, 1.40], P=0.30) and the duration of ventilation (mean difference[MD] -0.30, 95% CI [-0.64, 0.04], P=0.09), ICU stay (MD -0.38, 95% CI [-1.03, 0.27], P=0.25), and hospital stay (MD -0.56, 95% CI [-1.44, 0.32], P=0.22), high PEEP indeed increased the level of PaO2/FIO2 (MD 32.39, 95% CI [13.06, 51.72], P=0.001), and therefore decreased the incidences of ARDS (RR 0.57, 95% CI [0.37, 0.89], P=0.01) and hypoxaemia (RR 0.60, 95% CI [0.41, 0.88], P=0.009). In addition, although total results did not reveal the advantage of high PEEP on other secondary outcomes regarding atelectasis, barotrauma, ventilator associated pneumonia (VAP), hypotension, mean arterial pressure (MAP), SaO2 and lactate, subgroup analysis seemed to obtain different results. The TSA results suggested more RCTs were needed. Conclusion: Ventilation with high PEEP in ICU patients without ARDS may improve the level of oxygenation (PaO2/FIO2) resulting in low incidences of ARDS and hypoxaemia. Nevertheless, other clinical outcomes including in-hospital and 28-day mortality, duration of ventilation, ICU stay and hospital stay, pulmonary complications, hemodynamics and post-operative fluid balance did not show any significant difference.