The effects of hydrogen sulfide (H2S) on chilling injury (CI), reactive oxygen species (ROS) metabolism, sugar metabolism, pentose phosphate pathway (PPP), and membrane lipid metabolism in loquat fruit throughout the refrigerated period were investigated in this study. The findings indicated that H2S application restrained the increase in internal browning (IB), malondialdehyde (MDA) content, and electrolyte leakage, while sustaining higher total phenolic and total flavonoid levels, and lower soluble quinone content in loquat fruit. Besides, H2S promoted antioxidant accumulation and increased antioxidant enzyme activities by the regulation of ROS metabolism, along with increasing fructose and glucose levels and reducing power by activating sugar metabolism and PPP. Furthermore, H2S treatment retarded the degradation of phospholipids and fatty acids in loquat fruit by modulating membrane lipid metabolism relevant enzyme activities. These findings indicated that H2S application mitigated CI in loquat fruit by alleviating oxidative stress and maintaining cell membrane structural integrity.