OBJECTIVE The present study evaluated the nephroprotective effects of anhuienoside C (AC) against diabetic nephropathy (DN) in rats. METHODS Diabetic nephropathy was induced by administration of a high-fat diet (HFD) for 8 weeks and intraperitoneal administration of streptozotocin (STZ; 30 mg/kg) at the end of the fourth week of this protocol. Effects of AC on blood glucose levels, renal function markers, and mediators of inflammation in the serum of DN rats were assessed. RESULTS Anhuienoside C treatment reduced the blood glucose levels and attenuated the increased levels of renal injury markers in DN rats. Anhuienoside C also increased podocyte counts; alleviated the changes in podocin, desmin, and nephrin protein levels; and ameliorated the altered pathophysiology in the kidney tissues induced by DN. Compared with the DN group, the levels of inflammatory markers and mediators of oxidative stress were reduced in the serum and kidney tissues of the AC-treated groups. Moreover, treatment with AC ameliorates the altered expression of podocin, nephrin, and desmin proteins in the renal tissue of HFD/STZ-induced kidney-injured rats. CONCLUSIONS In conclusion, AC protected against podocyte injury by regulating nuclear factor kappa-light-chain-enhancer of activated B cells/protein kinase B pathway in a rat model of DN.