Human amniotic mesenchymal stem cell-derived exosome inhibits ferroptosis through the suppression of JNK/MAPK-mediated EGR1 expression in spontaneous intracerebral hemorrhage

INTRODUCTION: Spontaneous intracerebral hemorrhage (ICH) represents an essential part of stroke, wherein ferroptosis is identified as a critical factor leading to the pathogenesis. Symptomatical treatments are mainly adopted which tend to leave severe neurological deficits. Recently, human amniotic mesenchymal stem cell (hAMSC) therapy has developed rapidly in the field of regenerative medicine, with its secreted exosomes (hAMSC-Exos) regarded as effective alternatives owing to the ability to penetrate the blood-brain barrier. Therefore, we aim to explore the therapeutic efficacy as well as molecular mechanism of hAMSC-Exos in ICH models. METHODS: Firstly, hAMSCs and their exosomes were extracted and characterized. Subsequently, the impact of hAMSC-Exos was assessed by evaluating the restoration of neurological function in ICH rat models induced by collagenase VII and in PC12 cells induced by hemin. Moreover, transcriptome sequencing of PC12 cell models identified the potential hub genes and pathways, which were subsequently verified through the western blot among the groups and further examined by the overexpression plasmid transfection and pathway activator treatment. RESULTS: hAMSC-Exos were successfully obtained and characterized. Afterwards, the in vivo and in vitro functional experiments demonstrated the effectiveness of hAMSC-Exos therapy in ICH models by reducing the level of ferroptosis, apoptosis and inflammatory factors and restoring the neurological function. Transcriptome sequencing identified the hub gene Egr1 and the key pathway MAPK, which were subsequently verified by the western blot. Specifically, hAMSC-Exos reduced the elevated EGR1 expression and p-JNK/JNK ratio in the ICH model. However, EGR1 overexpression plasmid transfection and anisomycin treatment reactivated ferroptosis in hAMSC-Exos-treated ICH models. Notably, the expression of EGR1 was increased following anisomycin administration. CONCLUSION: The hAMSC-Exos may attenuate post-ICH ferroptosis and restore neurological function via down-regulating EGR1 mediated by JNK/MAPK in the rat and PC12 cell model of ICH, which might provide new approaches and drug targets in the management of the disease.