Objective: To investigate the renal protective effect of Xuezhikang on rats with diabetic kidney disease (DKD) through adenosine 5‘-monophosphate-activated protein kinase(AMPK)/endothelial nitric oxide synthase (eNOS). Method SD rats were randomly divided into the control group, the model group (establishing a DKD model), the Xuezhikang group (DKD modeling and 1200mg/kg/d Xuezhikang gavage), the inhibitor group (DKD modeling and 0.2 mg/kg/d AMPK inhibitor Compound intraperitoneal injection), the Xuezhikang+inhibitor group(DKD modeling and 1200mg/kg/d Xuezhikang gavage, 0.2 mg/kg/d AMPK inhibitor Compound C intraperitoneal injection). After 4 weeks of intervention, 24-hour urine albumin, serum creatinine (Scr), blood urea nitrogen (BUN), and renal pathological changes, the expression levels of phosphorylated AMPK (p-AMPK) and eNOS, as well as the levels of tumor necrosis factor -α (TNF-α), interleukin-6 (IL-6), superoxide dismutase (SOD), and malondialdehyde (MDA) in the kidneys were detected. Result: Compared with the model group, the renal pathological changes were alleviated, the levels of 24-hour urinary albumin, Scr, BUN, TNF-α, IL-6, and MDA in the kidneys decreased, while the expression levels of p-AMPK, eNOS and the level of SOD in the kidneys increased in the Xuezhikang group(P<0.05); the renal pathological changes were aggravated, the levels of 24-hour urinary albumin, Scr, BUN, TNF-α, IL-6 and MDA in the kidneys increased, while the expression levels of p-AMPK, eNOS and the level of SOD in the kidneys decreased in the inhibitor group(P<0.05). Compared with the Xuezhikang group, the renal pathological changes were aggravated, the levels of 24-hour urinary albumin, Scr, BUN, TNF-α, IL-6 and MDA in the kidneys increased, while the expression levels of p-AMPK, eNOS and the level of SOD in the kidneys decreased in the Xuezhikang + inhibitor group(P<0.05). Conclusion: Xuezhikang alleviates renal injury, inflammatory response and oxidative stress response in DKD rats by activating AMPK/eNOS.