AIM: To evaluate the sedative effect of remimazolam tosilate and its impact on iatrogenic stress and immune function biomarkers, in order to explore its feasibility and potential advantages as a novel sedative for mechanically ventilated Intensive Care Unit (ICU) patients. METHODS: In a retrospective analysis, 136 ICU patients who were admitted to the Second Affiliated Hospital of Jiaxing University between March and December 2022 and received mechanical ventilation were enrolled and divided into two groups based on the sedation protocol during intubation: conventional propofol group (Control group,n= 79) and remimazolam tosilate group (Remi group,n= 57). The sedation success rate and adverse reaction rate were recorded. The iatrogenic stress markers including epinephrine (E), norepinephrine (NE), and cortisol (Cor) and the T lymphocyte subpopulation including CD3+T%, CD4+T%, CD8+T% were determined using enzyme-linked immunosorbent assay and flow cytometry assay before anesthesia induction (T0), at one week (T4), and at one month after anesthesia (T5). RESULTS: The sedation success rate was significantly higher in the Remi group (80% [50%, 88%]) compared to the Control group (72% [43%, 85%]) (p< 0.05). The Remi group exhibited significantly lower Cor levels at T4 and lower E, NE, and Cor levels at T5 compared to the Control group (p< 0.05). The Remi group exhibited significantly higher CD4+T% levels at T4 and lower CD8+T% levels at T5 time points compared to the Control group (p< 0.05). No significant difference was found in the overall incidence of adverse events between the two groups (p> 0.05). CONCLUSIONS: Compared with propofol, remimazolam tosilate may offer potential benefits in mitigating iatrogenic stress and preserving immune homeostasis, while ensuring adequate sedation during ICU mechanical ventilation. It could be a promising sedative agent for use in the ICU setting.