Abstract Painful diabetic peripheral neuropathy (PDPN) is a multifactorial disabling complication of diabetes, yet the pathogenesis involving mitochondrial dysfunction in immune cells remains unclear. This study aimed to identify mitochondria-related genes (MRGs) in PDPN and explore their biological relevance. Based on single-cell transcriptomic analysis, we identified three key MRGs (Hmgcs2, Nudt5, and Cpt1c) shared across two independent datasets and validated them in rat PDPN models.