Diabetic kidney disease (DKD) is one of the leading causes of end-stage renal disease. Verbascoside is a widespread phenylethanoid glycoside with potent anti-inflammatory, antioxidant and hypoglycemic properties. And this study aimed to explore the renoprotective effect of verbascoside against DKD with the underlying mechanism. After administration of verbascoside for four consecutive weeks, fast blood glucose, albumin-creatinine ratio and podocyte damage in diabetic mice were alleviated, especially at the dose of 150mg/kg/d. Moreover, inflammatory response, cell apoptosis and autophagy were improved dose-dependently in the kidneys of diabetic mice and high glucose-stimulated podocytes. Mechanistically, verbascoside reversed the elevated NR4A1 expression and suppressed LKB1 to inhibit AMPKα phosphorylation. Silencing NR4A1 could inhibit the LKB1 and phospho-AMPKα expression, and it could relieve stress response in injured podocytes. Collectively, our results indicated that verbascoside alleviated podocyte injury of DKD via regulating the NR4A1-LKB1-AMPK signaling.