Objective:To investigate the therapeutic effects and mechanisms of human mesenchymal stem cell-derived exosomes(hMSCs-Exo)carrying the NGF gene in treating ischemic stroke in rats,aiming to provide new insights and tr...Objective:To investigate the therapeutic effects and mechanisms of human mesenchymal stem cell-derived exosomes(hMSCs-Exo)carrying the NGF gene in treating ischemic stroke in rats,aiming to provide new insights and treatment methods for ischemic stroke therapy.Methods:After successful construction of the cerebral ischemia model in 40 male SPF-grade SD rats aged 6-8 weeks,the model rats were randomly divided into 4 groups:Sham group,PBS group,hMSCs-Exo group,and NGF-hMSCs-Exo group,with 10 rats in each group.The rat MCAO model was prepared using the classic filament method,and NGF-hMSCs-Exo were injected via the tail vein into the MCAO model rats.The expression of the NGF gene in brain ischemic tissues,neuronal regeneration,and rat neurological function recovery were observed using TTC staining,memory function evaluation,Western blot,qRT-PCR,and other methods.Results:Compared with the Sham group,neurological deficits were significant in the PBS group(P<0.01).Compared with the PBS group,neurological scores improved in the hMSCs-Exo group and NGF-hMSCs-Exo group(P<0.05).Compared with the hMSCs-Exo group,the improvement in neurological deficits was more significant in the NGF-hMSCs-Exo group(P<0.05).The infarct area after NGF-hMSCs-Exo intervention was significantly reduced(P<0.05)compared with the Sham group.Compared with the PBS group,relative expression levels of NGF mRNA and protein decreased,while Caspase-3 mRNA and protein expression significantly increased in the PBS group(P<0.01).Compared with the PBS group and hMSCs-Exo group,there were differences in NGF and Caspase-3 mRNA and protein expression in the NGF-hMSCs-Exo group rat brain tissues(P<0.05).Conclusion:Treatment with human mesenchymal stem cell-derived exosomes carrying the NGF gene improves cognitive function and exerts protective effects on SD rats while inhibiting apoptotic levels in cells.展开更多
Objective Our previous studies established that microRNA(miR)-451 from human umbilical cord mesenchymal stem cell-derived exosomes(hUC-MSC-Exos)alleviates acute lung injury(ALI).This study aims to elucidate the mechan...Objective Our previous studies established that microRNA(miR)-451 from human umbilical cord mesenchymal stem cell-derived exosomes(hUC-MSC-Exos)alleviates acute lung injury(ALI).This study aims to elucidate the mechanisms by which miR-451 in hUC-MSC-Exos reduces ALI by modulating macrophage autophagy.Methods Exosomes were isolated from hUC-MSCs.Severe burn-induced ALI rat models were treated with hUC-MSC-Exos carrying the miR-451 inhibitor.Hematoxylin-eosin staining evaluated inflammatory injury.Enzyme-linked immunosorbnent assay measured lipopolysaccharide(LPS),tumor necrosis factor-α,and interleukin-1βlevels.qRT-PCR detected miR-451 and tuberous sclerosis complex 1(TSC1)expressions.The regulatory role of miR-451 on TSC1 was determined using a dual-luciferase reporter system.Western blotting determined TSC1 and proteins related to the mammalian target of rapamycin(mTOR)pathway and autophagy.Immunofluorescence analysis was conducted to examine exosomes phagocytosis in alveolar macrophages and autophagy level.Results hUC-MSC-Exos with miR-451 inhibitor reduced burn-induced ALI and promoted macrophage autophagy.MiR-451 could be transferred from hUC-MSCs to alveolar macrophages via exosomes and directly targeted TSC1.Inhibiting miR-451 in hUC-MSC-Exos elevated TSC1 expression and inactivated the mTOR pathway in alveolar macrophages.Silencing TSC1 activated mTOR signaling and inhibited autophagy,while TSC1 knockdown reversed the autophagy from the miR-451 inhibitor-induced.Conclusion miR-451 from hUC-MSC exosomes improves ALI by suppressing alveolar macrophage autophagy through modulation of the TSC1/mTOR pathway,providing a potential therapeutic strategy for ALI.展开更多
基金Clinical Medicine Science and Technology Development Fund of Jiangsu University 2021(Natural Science Category)(Grant No.JLY2021004)。
文摘Objective:To investigate the therapeutic effects and mechanisms of human mesenchymal stem cell-derived exosomes(hMSCs-Exo)carrying the NGF gene in treating ischemic stroke in rats,aiming to provide new insights and treatment methods for ischemic stroke therapy.Methods:After successful construction of the cerebral ischemia model in 40 male SPF-grade SD rats aged 6-8 weeks,the model rats were randomly divided into 4 groups:Sham group,PBS group,hMSCs-Exo group,and NGF-hMSCs-Exo group,with 10 rats in each group.The rat MCAO model was prepared using the classic filament method,and NGF-hMSCs-Exo were injected via the tail vein into the MCAO model rats.The expression of the NGF gene in brain ischemic tissues,neuronal regeneration,and rat neurological function recovery were observed using TTC staining,memory function evaluation,Western blot,qRT-PCR,and other methods.Results:Compared with the Sham group,neurological deficits were significant in the PBS group(P<0.01).Compared with the PBS group,neurological scores improved in the hMSCs-Exo group and NGF-hMSCs-Exo group(P<0.05).Compared with the hMSCs-Exo group,the improvement in neurological deficits was more significant in the NGF-hMSCs-Exo group(P<0.05).The infarct area after NGF-hMSCs-Exo intervention was significantly reduced(P<0.05)compared with the Sham group.Compared with the PBS group,relative expression levels of NGF mRNA and protein decreased,while Caspase-3 mRNA and protein expression significantly increased in the PBS group(P<0.01).Compared with the PBS group and hMSCs-Exo group,there were differences in NGF and Caspase-3 mRNA and protein expression in the NGF-hMSCs-Exo group rat brain tissues(P<0.05).Conclusion:Treatment with human mesenchymal stem cell-derived exosomes carrying the NGF gene improves cognitive function and exerts protective effects on SD rats while inhibiting apoptotic levels in cells.
基金supported by the tenth batch of"3221"industrial innovation and scientific research projects in Bengbu City(beng talent[2020]No.8)the 2021 Bengbu Medical College Science and Technology Project[Natural Science,Project Number:2021byzd217].
文摘Objective Our previous studies established that microRNA(miR)-451 from human umbilical cord mesenchymal stem cell-derived exosomes(hUC-MSC-Exos)alleviates acute lung injury(ALI).This study aims to elucidate the mechanisms by which miR-451 in hUC-MSC-Exos reduces ALI by modulating macrophage autophagy.Methods Exosomes were isolated from hUC-MSCs.Severe burn-induced ALI rat models were treated with hUC-MSC-Exos carrying the miR-451 inhibitor.Hematoxylin-eosin staining evaluated inflammatory injury.Enzyme-linked immunosorbnent assay measured lipopolysaccharide(LPS),tumor necrosis factor-α,and interleukin-1βlevels.qRT-PCR detected miR-451 and tuberous sclerosis complex 1(TSC1)expressions.The regulatory role of miR-451 on TSC1 was determined using a dual-luciferase reporter system.Western blotting determined TSC1 and proteins related to the mammalian target of rapamycin(mTOR)pathway and autophagy.Immunofluorescence analysis was conducted to examine exosomes phagocytosis in alveolar macrophages and autophagy level.Results hUC-MSC-Exos with miR-451 inhibitor reduced burn-induced ALI and promoted macrophage autophagy.MiR-451 could be transferred from hUC-MSCs to alveolar macrophages via exosomes and directly targeted TSC1.Inhibiting miR-451 in hUC-MSC-Exos elevated TSC1 expression and inactivated the mTOR pathway in alveolar macrophages.Silencing TSC1 activated mTOR signaling and inhibited autophagy,while TSC1 knockdown reversed the autophagy from the miR-451 inhibitor-induced.Conclusion miR-451 from hUC-MSC exosomes improves ALI by suppressing alveolar macrophage autophagy through modulation of the TSC1/mTOR pathway,providing a potential therapeutic strategy for ALI.
