Autophagy is a ubiquitous and fundamental catabolic vital process for maintaining cellular homeostasis, achieved by degrading and recycling cytoplasmic components, particularly under conditions of nutrient deprivation or metabolic stress. This mechanism is also integral to the selective clearance of misfolded or aggregated proteins, the removal of dysfunctional organelles (such as mitochondria and the endoplasmic reticulum), and the intracellular degradation of pathogens, including those associated with peroxisomes. In this study, we screened and identified sesamin, a bioactive compound isolated from Hypericum hookerianum extracts, as a novel autophagy activator. Our results demonstrated that sesamin effectively induces autophagy and activates the lysosome biogenesis pathway.