Mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) are implicated in cerebral ischemia reperfusion (I/R) injury process. In this study, after extraction and identification of human umbilical cord MSCs (HMCs)-derived EVs, I/R rat models were established and treated with HMC-EVs to measure pathological damage, apoptosis and inflammation in brain tissues. The differentially expressed microRNAs (miRs) in HMC-EVs and I/R rat tissues were screened. The downstream gene and pathways of miR-24 were analyzed. The gain- and loss-of function of miR-24 in HMC-EVs was performed in I/R rat models and hypoxia/reoxygenation (H/R) cell models. SH-SY5Y cells were subjected to hypoxia and biological behaviors were detected by MTT assay, colony formation experiment, EdU staining and Transwell assays, and cells were incubated with the inhibitors of downstream pathways. As expected, infarct size, brain tissue apoptosis and inflammation were decreased after HMC-EVs treatment. miR-24 overexpression in HMC-EVs reduced I/R injury, while miR-24 knockdown in HMC-EVs impaired the protective roles of HMC-EVs in I/R injury. HMC-EVs-carried miR-24 could target AQP4 to activate the P38 MAPK/ERK1/2/P13K/AKT pathway, and thus promoted the proliferation and migration of SH-SY5Y cells after H/R injury, which were reversed by LY294002 and PD98095. Taken together, HMC-EVs-carried miR-24 played protective roles in I/R injury, possibly by targeting AQP4 and activating the P38 MAPK/ERK1/2/P13K/AKT pathway. This study may offer novel perspective for I/R injury treatment.