【摘要】 目的 分析烧伤亚急性期单核细胞表面抗原递呈因子CD86和HLA-DR的表达变化，并探讨单核细胞功能障碍触发烧伤后持续炎症的分子机制。方法 将小鼠分为假烧伤组和烧伤组，并建立30%体表面烧伤小鼠模型，取烧伤第8天的小鼠脾脏，分离单核细胞后，进行转录组学分析；构建Notch1慢病毒转染小鼠模型，制作假烧伤+空白对照组(sham-NC)、烧伤+空白对照组(burn-NC)、假烧伤+Notch1 sh组(sham-sh)、烧伤+Notch1 sh(burn-sh)模型。采用qRT-PCR、Western blot检测单核细胞表面抗原CD86和HLA-DR的表达；通过免疫荧光分析脾脏单核细胞CD86和HLA-DR的表达；ELISA检测血清中IL-10、CRP的含量。结果 烧伤第8天，与假烧伤组对比，烧伤组单核细胞表面抗原CD86和HLA-DR表达显著下降，血清IL-10、CRP含量增加，且IL-10、CRP的含量分别与CD86、HLA-DR的表达呈明显的负相关性。转录组学分析显示，烧伤组脾脏单核细胞中有258种基因显著上调，360种基因显著下调；其中差异基因主要富集于Notch信号通路；Western blot结果显示Notch1表达显著上调。以慢病毒抑制小鼠体内Notch1信号后，与burn-NC对比，burn-sh组单核细胞表面CD86和HLA-DR表达增加，血清IL-10、CRP含量下降。结论 Notch1信号介导烧伤亚急性期单核细胞功能抑制，抑制体内Notch1信号改善烧伤后单核细胞抗原递呈能力，并改善炎症因子分泌。更多还原

【Abstract】 Objective To analyze the expression of monocyte surface antigen-presenting factors CD86⁺ and HLA-DR in the subacute phase of burn injury, and the molecular mechanism of monocytes contributing to persistent inflammation.Methods The mice were divided into the sham group and the burn group, and a mouse model of 30% body surface burn was established.Spleens were collected from mice on the 8th day after burn injury, and monocytes were isolated for transcriptomic analysis.A mouse model of Notch1 lentiviral transfection was constructed, and models of sham burn + blank control（sham-NC）,burn + blank control（burn-NC）,sham burn + Notch1 shRNA（sham-sh）,and burn + Notch1 shRNA（burn-sh）were established.The qRT-PCR and Western blot were used to detect the expressions of monocyte surface antigens CD86⁺ and HLA-DR.Immunofluorescence was performed to analyze the expression of CD86⁺ and HLA-DR in splenic monocytes.ELISA was used to analyze serum levels of IL-10 and CRP.Results On the 8th day after burn injury, compared with the sham burn group, the expressions of monocyte surface antigens CD86⁺ and HLA-DR were decreased, while serum levels of IL-10 and CRP were increased in the burn group.Contents of IL-10 and CRP showed significant negative correlations with CD86⁺ and HLA-DR,respectively.Transcriptomic analysis revealed 258 significantly upregulated genes and 360 significantly downregulated genes in splenic monocytes from the burn group.The differentially expressed genes primarily enriched in the Notch signaling pathway.Western blot results showed a significant upregulation of Notch1 expression.After lentiviral inhibition of Notch1 signaling in vivo,compared with the burn-NC group, expressions of CD86⁺ and HLA-DR on monocyte surfaces were increased, and serum levels of IL-10 and CRP were decreased in the burn-sh group.Conclusion The functional inhibition of splenic monocytes is mediated by Notch1 signaling pathway in the subacute phase of burn injury.Inhibition of Notch1 signaling in vivo improves the antigen-presenting capacity of monocytes after burn injury and inhibits the secretion of inflammatory factors.更多还原