<p>This study investigates the specific mechanisms by which SIRT1/SIRT3 modulates the reduction of mitochondrial regeneration and the inflammatory response in cerebral ischemia-reperfusion(I/R) injury exacerbated by hyperglycemia. A streptozotocin-induced diabetic rat model was developed to simulate diabetes mellitus(DM), and embolization of the middle cerebral artery was performed to establish cerebral I/R injury. Brain damage was assessed via neurological function scores, TTC staining, HE staining and electron microscopy. SIRT1/SIRT3 mRNA levels and initial mitochondrial DNA (mtDNA) content were quantified using RT-qPCR. Proteins of SIRT1/SIRT3 and proteins associated with mitochondrial regeneration and the inflammatory response were validated by western blot analysis. The production of reactive oxygen species (ROS) and inflammatory factors was assayed by ELISA. The findings indicated a significant increase in neurological function scores across all groups experiencing diabetic cerebral I/R injury (<i>P</i> &lt; 0.05). Additionally, a notable increase was observed in cerebral infarction volume (<i>P</i> &lt; 0.05) and neuronal necrosis (<i>P</i> &lt; 0.05) and the structure and morphology of the cells were significantly altered. the levels of SIRT1/SIRT3 were found to be significantly reduced (<i>P</i> &lt; 0.05). Furthermore, the levels of PGC-1α, NRF1, and TFAM, which are linked to mitochondrial regeneration, were found to be significantly reduced (<i>P</i> &lt; 0.05). Additionally, ROS and their activities, as well as the inflammasome NLRP3 and inflammatory factors such as TNF-α, IL-6, and IL-1β, were significantly increased (<i>P</i> &lt; 0.05). These findings suggest that diabetes exacerbates I/R injury in the rat brain by further hindering mitochondrial regeneration, possibly through the SIRT1/SIRT3-PGC-1α-NRF1-TFAM pathway. Additionally, diabetes intensifies this injury by amplifying the inflammatory cascade response, which may be associated with the SIRT1/SIRT3-ROS-NLRP3 pathway. </p>