Objective To investigate the mechanism of troxerutin (TRO) alleviating renal injury in rats with adriamycin (ADM)-induced nephrotic syndrome (NS) through autophagy mediated by AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway. Methods ADM-induced NS rat model was established. Sixty SPF SD rats were randomly grouped into Control group, Model group, low-dose TRO group (TRO-L group, 50 mg/kg), high-dose TRO group (TRO-H group, 100 mg/kg), and high-dose TRO+AMPK inhibitor Compound C group (TRO-H+CC group, 100 mg/kg TRO+20 mg/kg CC), 12 animals in each group. The 24-hour urine of the rats was collected before and after modeling to detect the urinary protein content. The contents of serum albumin (ALB), total protein (TP), triacylglycerol (TG), cholesterol (TC), blood urea nitrogen (BUN) and serum creatinine (Scr) in rat serum were detected by automatic biochemical analyzer. HE staining was applied to observe the histopathological changes of the rat kidneys. ELISA method was applied to detect the content of SOD and MDA in kidney tissue of rats. TUNEL staining was applied to observe apoptosis in rat kidney tissue. RT-qPCR method was applied to detect the mRNA expression levels of AMPK and mTOR in rat kidney tissue; Western Blot was applied to detect the expression of AMPK/mTOR pathway-related proteins in rat kidney tissue. Results Compared with the control group, the 24-hour urine protein content, TG, TC, BUN and Scr levels, renal tissue apoptosis rate, mTOR mRNA expression and p-mTOR/mTOR were obviously increased, the serum ALB, TP levels, renal SOD level, AMPK mRNA expression and p-AMPK/AMPK were obviously decreased in the Model group (P<0.05), renal tissue pathological damage was severe.Compared with the Model group, the TRO-L and TRO-H groups had reduced renal pathological damage and decreased cell apoptosis, and the trends of related indicators were opposite to the above (P<0.05). Compound C attenuated the protective effect of TRO on kidney function in NS rats (P<0.05). Conclusion TRO alleviates ADM-induced kidney injury in NS rats by activating the autophagy response mediated by the AMPK/mTOR pathway