Sepsis is a life-threatening disorder that develops in response to an infection. Kidneys are one of the most common organs affected by sepsis. This study explored the influence of Sestrin2 regulating NLRP3 inflammasome activation on sepsis-induced renal injury. A sepsis mouse model was established by intraperitoneal injection of LPS. Adenoviral transduction was used to upregulate Sestrin2 expression. H&E staining, TUNEL staining, DHE staining, ELISA, RT-PCR, and western blotting were applied to the detections of pathological changes, ROS, oxidative stress markers, Sestrin2 expression, apoptosis-related proteins, and NLRP3 inflammasome-related factors in renal tissues. A HK-2 cell model of LPS-induced injury was constructed, and the release of IL-6 and TNF-α and expression of NLRP3 were detected using ELISA and immunofluorescence, respectively. Sestrin2 overexpression suppressed inflammation and reduced renal injury and apoptosis in LPS-treated mice. Moreover, overexpressed Sestrin2 resulted in inhibited levels of ROS, NLRP3, caspase-1, and IL-1β in renal tissues, as well as reduced TNF-α and IL-6 secretion and NLRP3 expression in LPS-stimulated HK-2 cells. Collectively, Sestrin2 can inhibit inflammation and reduce sepsis-induced renal injury, which may be associated with NLRP3 inflammasome inactivation and oxidative stress reduction.