Astrocytes and microglia play an orchestrated role following spinal cord injury;however,the molecular mechanisms through which microglia regulate astrocytes after spinal cord injury are not yet fully understood.Herein...Astrocytes and microglia play an orchestrated role following spinal cord injury;however,the molecular mechanisms through which microglia regulate astrocytes after spinal cord injury are not yet fully understood.Herein,microglia were pharmacologically depleted and the effects on the astrocytic response were examined.We further explored the potential mechanisms involving the signal transducers and activators of transcription 3(STAT3)pathway.For in vivo experiments,we constructed a contusion spinal cord injury model in C57BL/6 mice.To deplete microglia,all mice were treated with colony-stimulating factor 1 receptor inhibitor PLX3397,starting 2 weeks prior to surgery until they were sacrificed.Cell proliferation was examined by 5-ethynyl-2-deoxyuridine(EdU)and three pivotal inflammatory cytokines were detected by a specific Bio-Plex Pro^(TM) Reagent Kit.Locomotor function,neuroinflammation,astrocyte activation and phosphorylated STAT3(pSTAT3,a maker of activation of STAT3 signaling)levels were determined.For in vitro experiments,a microglia and astrocyte coculture system was established,and the small molecule STA21,which blocks STAT3 activation,was applied to investigate whether STAT3 signaling is involved in mediating astrocyte proliferation induced by microglia.PLX3397 administration disrupted glial scar formation,increased inflammatory spillover,induced diffuse tissue damage and impaired functional recovery after spinal cord injury.Microglial depletion markedly reduced EdU+proliferating cells,especially proliferating astrocytes at 7 days after spinal cord injury.RNA sequencing analysis showed that the JAK/STAT3 pathway was downregulated in mice treated with PLX3397.Double immunofluorescence staining confirmed that PLX3397 significantly decreased STAT3 expression in astrocytes.Importantly,in vitro coculture of astrocytes and microglia showed that microglia-induced astrocyte proliferation was abolished by STA21 administration.These findings suggest that microglial depletion impaired astrocyte proliferation and astro展开更多
目的:体外培养人脂肪干细胞(adi pose-deri ved st em cel l s,ADSCs)并行EdU标记,并通过测试标记率及标记后对细胞增殖分化的影响确定优化的标记时间及浓度组合。方法:使用分别采5μM,10μM,20μM,50μM的浓度及12h,24h进行标记标记AD...目的:体外培养人脂肪干细胞(adi pose-deri ved st em cel l s,ADSCs)并行EdU标记,并通过测试标记率及标记后对细胞增殖分化的影响确定优化的标记时间及浓度组合。方法:使用分别采5μM,10μM,20μM,50μM的浓度及12h,24h进行标记标记ADSC,并以流式细胞仪精确测定标记率。挑选出各12h及24h时优化标记浓度,并进行标记后测定细胞活性,增殖及诱导分化实验。结果:P3代细胞约90%以上表达表面标记CD90+,CD105+,CD44+。基本不表达细胞表面标记CD45+,CD35+。通过各试验,得出最适浓度组合10μM,12h,对进一步研究脂肪干细胞在辅助脂肪移植中的作用具有指导意义。结论:EdU是标记方法简单有效,体外最佳标记浓度组合是10μM,12h。展开更多
Community education is an essential carrier of continuing education and plays a positive role in promoting continuing education of migrant workers. On the one hand,it can raise employment quality and labor skills of m...Community education is an essential carrier of continuing education and plays a positive role in promoting continuing education of migrant workers. On the one hand,it can raise employment quality and labor skills of migrant workers; on the other hand,it manifests function of serving society of community education. Besides,it is also an important measure for building learning society and lifelong learning system.From the perspective of interactive development,it discusses interactive relationship between community education and migrant workers' continuing education,analyzes their interactive mechanism,and comes up with recommendations for developing community education and migrant workers' continuing education.展开更多
基金supported by the Natural Science Foundation of Guangdong Province,No.2020A1515010090(to ZLZ)the Science and Technology Project Foundation of Guangzhou City,No.202002030004(to HZ).
文摘Astrocytes and microglia play an orchestrated role following spinal cord injury;however,the molecular mechanisms through which microglia regulate astrocytes after spinal cord injury are not yet fully understood.Herein,microglia were pharmacologically depleted and the effects on the astrocytic response were examined.We further explored the potential mechanisms involving the signal transducers and activators of transcription 3(STAT3)pathway.For in vivo experiments,we constructed a contusion spinal cord injury model in C57BL/6 mice.To deplete microglia,all mice were treated with colony-stimulating factor 1 receptor inhibitor PLX3397,starting 2 weeks prior to surgery until they were sacrificed.Cell proliferation was examined by 5-ethynyl-2-deoxyuridine(EdU)and three pivotal inflammatory cytokines were detected by a specific Bio-Plex Pro^(TM) Reagent Kit.Locomotor function,neuroinflammation,astrocyte activation and phosphorylated STAT3(pSTAT3,a maker of activation of STAT3 signaling)levels were determined.For in vitro experiments,a microglia and astrocyte coculture system was established,and the small molecule STA21,which blocks STAT3 activation,was applied to investigate whether STAT3 signaling is involved in mediating astrocyte proliferation induced by microglia.PLX3397 administration disrupted glial scar formation,increased inflammatory spillover,induced diffuse tissue damage and impaired functional recovery after spinal cord injury.Microglial depletion markedly reduced EdU+proliferating cells,especially proliferating astrocytes at 7 days after spinal cord injury.RNA sequencing analysis showed that the JAK/STAT3 pathway was downregulated in mice treated with PLX3397.Double immunofluorescence staining confirmed that PLX3397 significantly decreased STAT3 expression in astrocytes.Importantly,in vitro coculture of astrocytes and microglia showed that microglia-induced astrocyte proliferation was abolished by STA21 administration.These findings suggest that microglial depletion impaired astrocyte proliferation and astro
文摘目的:体外培养人脂肪干细胞(adi pose-deri ved st em cel l s,ADSCs)并行EdU标记,并通过测试标记率及标记后对细胞增殖分化的影响确定优化的标记时间及浓度组合。方法:使用分别采5μM,10μM,20μM,50μM的浓度及12h,24h进行标记标记ADSC,并以流式细胞仪精确测定标记率。挑选出各12h及24h时优化标记浓度,并进行标记后测定细胞活性,增殖及诱导分化实验。结果:P3代细胞约90%以上表达表面标记CD90+,CD105+,CD44+。基本不表达细胞表面标记CD45+,CD35+。通过各试验,得出最适浓度组合10μM,12h,对进一步研究脂肪干细胞在辅助脂肪移植中的作用具有指导意义。结论:EdU是标记方法简单有效,体外最佳标记浓度组合是10μM,12h。
基金Supported by Project for Twelfth Five-Year Plan of National Education Information Technology Research in 2013 "Exploration and Practice of Constructing Network Course System for the Open University"(136241485)Scientific Research Planning Project of the Chinese Society of Vocational and Technical Education in 2012-2013 "Study on Interactive Development of Community Education and Migrant Workers'Continuing Education from the Perspective of Human Capital"(574714)
文摘Community education is an essential carrier of continuing education and plays a positive role in promoting continuing education of migrant workers. On the one hand,it can raise employment quality and labor skills of migrant workers; on the other hand,it manifests function of serving society of community education. Besides,it is also an important measure for building learning society and lifelong learning system.From the perspective of interactive development,it discusses interactive relationship between community education and migrant workers' continuing education,analyzes their interactive mechanism,and comes up with recommendations for developing community education and migrant workers' continuing education.
