Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we fo...Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we found that exosomal miR-486-3p is involved in the pathophysiology of SAH.Targeted delivery of miR-486-3p without blood-brain barrier(BBB)restriction to alleviate SAH is a promising neuroinflammation approach.Methods In this study,we modified exosomes(Exo)to form an RVG-miR 486-3p Exo(Exo/miR)to achieve targeted delivery of miR-486-3p to the brain.Neurological scores,brain water content,BBB damage,flow cytometry and FJC staining were used to determine the effect of miR-486-3p on SAH.Western blot analysis,ELISA and RT-qPCR were used to measure relevant protein and mRNA levels.Immunofluorescence staining and laser confocal detection were used to measure the expression of mitochondria,lysosomes and autophagosomes,and transmission electron microscopy was used to observe the level of mitophagy in the brain tissue of mice after SAH.Results Tail vein injection of Exo/miR improved targeting of miR-486-3p to the brains of SAH mice.The injection reduced levels of neuroinflammation-related factors by changing the phenotype switching of microglia,inhibiting the expression of sirtuin 2(SIRT2)and enhancing mitophagy.miR-486-3p treatment alleviated neurobehavioral disorders,brain oedema,BBB damage and neurodegeneration.Further research found that the mechanism was achieved by regulating the acetylation level of peroxisome proliferator-activated receptorγcoactivator l alpha(PGC-1α)after SIRT2 enters the nucleus.Conclusion Exo/miR treatment attenuates neuroinflammation after SAH by inhibiting SIRT2 expression and stimulating mitophagy,suggesting potential clinical applications.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.82271363(LY Wu),82301485(XX Chen),82201456(Y Zhou),82130037(CH.Hang))Natural Science Foundation of Jiangsu Province(No.2022JJ30060(Y Zhou))+1 种基金Anhui Provincial Department of Education Excellent Scientific Research and Innovation Team Project in Colleges and Universities(2022AH010073(NS Lai))Postgraduate Research&Practice Innovation Program of Jiangsu Province(SJCX24_0010(B Sheng)).
文摘Background Neuroinflammation participates in the pathogenesis of subarachnoid haemorrhage(SAH);however,no effective treatments exist.MicroRNAs regulate several aspects of neuronal dysfunction.In a previous study,we found that exosomal miR-486-3p is involved in the pathophysiology of SAH.Targeted delivery of miR-486-3p without blood-brain barrier(BBB)restriction to alleviate SAH is a promising neuroinflammation approach.Methods In this study,we modified exosomes(Exo)to form an RVG-miR 486-3p Exo(Exo/miR)to achieve targeted delivery of miR-486-3p to the brain.Neurological scores,brain water content,BBB damage,flow cytometry and FJC staining were used to determine the effect of miR-486-3p on SAH.Western blot analysis,ELISA and RT-qPCR were used to measure relevant protein and mRNA levels.Immunofluorescence staining and laser confocal detection were used to measure the expression of mitochondria,lysosomes and autophagosomes,and transmission electron microscopy was used to observe the level of mitophagy in the brain tissue of mice after SAH.Results Tail vein injection of Exo/miR improved targeting of miR-486-3p to the brains of SAH mice.The injection reduced levels of neuroinflammation-related factors by changing the phenotype switching of microglia,inhibiting the expression of sirtuin 2(SIRT2)and enhancing mitophagy.miR-486-3p treatment alleviated neurobehavioral disorders,brain oedema,BBB damage and neurodegeneration.Further research found that the mechanism was achieved by regulating the acetylation level of peroxisome proliferator-activated receptorγcoactivator l alpha(PGC-1α)after SIRT2 enters the nucleus.Conclusion Exo/miR treatment attenuates neuroinflammation after SAH by inhibiting SIRT2 expression and stimulating mitophagy,suggesting potential clinical applications.
