Objective To investigate the influence of hypericin (Hyp) on renal autophagy and the AMPK/ mTOR/ ULK1 pathway in nephrotic syndrome (NS) rats. Methods Thirty-two 6-week-old SD rats were grouped into normal (N), NS, Hyp (60 mg/ kg Hyp), and Hyp+CC (60 mg/ kg Hyp+0. 2 mg/ kg CC AMPK inhibitor) groups, with eight rats per group. The NS model was established by one-time injection of adriamycin (6.5 mg/ kg) through the tail vein, and the success rate of the model was 75. 0%. Rats in the Hyp group were given 60 mg/ kg Hyp intragastric administration; rats in the Hyp+CC group were given 60 mg/ kg Hyp intragastric administration and 0. 2 mg/ kg CC intraperitoneal injection; and rats in the N and NS groups were given the same amount of solvent once a day for 14 days. After administration, an automatic analyzer was applied to detect the levels of 24-h urine total protein (UTP), blood urea nitrogen (BUN), serum creatinine (Scr), and albumin (ALB). HE staining was used to observe the pathological morphology of the kidney tissue. The ultrastructure of the renal tissue was observed by transmission electron microscope. Western blot was applied to detect the expression of autophagy, podocyte, and AMPK/ mTOR/ ULK1 pathway proteins in the kidney. Immunofluorescence staining was applied to visualize the localization of autophagosomes and podocytes. Results Compared with the N group, the NS group had increased glomerular volume; atrophied or partially disappeared renal tubules; a thickened basement membrane; and obviously increased UTP, BUN and Scr levels, basement membrane thickness, foot process width, and p-AMPK/ AMPK ratio (P<0. 05), while the levels of ALB, LC3-II/ I, Beclin-1, Atg5, Atg7 and NPHS2; the relative fluorescence intensity of NPHS2 and Beclin-1; and p-AMPK/ AMPK and p-ULK1/ ULK1 ratios were obviously decreased (P<0. 05). Compared with the NS group, the Hyp treatment group had improved glomerular morphology and decreased UTP, BUN, and Scr levels, basement membrane thickness, foot process width, and p-AMPK/ AMPK (P<0. 05) ratio, but there was an increase in the protein levels of ALB, LC3-II/ I, Beclin-1, Atg5, Atg7, NPHS2; relative fluorescence intensity of NPHS2 and Beclin-1; and p-AMPK/ AMPK and p-ULK1/ ULK1 ratios (P<0. 05). Compared with the Hyp group, the Hyp+CC group’ s glomerular volume increased; renal tubules atrophied or partially disappeared; basement membrane thickened; and UTP, BUN, Scr levels, basement membrane thickness, foot process width, and p-AMPK/ AMPK ratio were obviously increased (P<0. 05), whereas the protein levels of ALB, LC3-II/ I, Beclin-1, Atg5, Atg7, and NPHS2; relative fluorescence intensity of NPHS2 and Beclin-1; and p-AMPK/ AMPK and p-ULK1/ ULK1 ratios were obviously decreased (P<0. 05). Conclusions Hyp may enhance the autophagic activity of renal cells and attenuate renal pathology, such as podocyte injury, in NS rats by activating the AMPK/ mTOR/ ULK1 pathway.