Ji Yabin, Chen Yiman, Tan Xixi, Huang Xiaowen, Gao Qiang, Ma Yinzhong, Yang Shilun, Yin Meifang, Yu Min, Fang Cheng, Wang Yu, Shi Zhu, Chang Junlei
Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
CNS Neurosci Ther. 2024 Apr;30(4):e14483. doi: 10.1111/cns.14483. Epub 2023 Oct 3.
Reperfusion therapy after ischemic stroke often causes brain microvascular injury. However, the underlying mechanisms are unclear.
Transcriptomic and proteomic analyses were performed on human cerebral microvascular endothelial cells following oxygen-glucose deprivation (OGD) or OGD plus recovery (OGD/R) to identify molecules and signaling pathways dysregulated by reperfusion. Major findings were further validated in a mouse model of cerebral ischemia and reperfusion.
Transcriptomic analysis identified 390 differentially expressed genes (DEGs) between the OGD/R and OGD group. Pathway analysis indicated that these genes were mostly associated with inflammation, including the TNF signaling pathway, TGF-β signaling pathway, cytokine-cytokine receptor interaction, NOD-like receptor signaling pathway, and NF-κB signaling pathway. Proteomic analysis identified 201 differentially expressed proteins (DEPs), which were primarily associated with extracellular matrix destruction and remodeling, impairment of endothelial transport function, and inflammatory responses. Six genes (DUSP1, JUNB, NFKBIA, NR4A1, SERPINE1, and THBS1) were upregulated by OGD/R at both the mRNA and protein levels. In mice with cerebral ischemia and reperfusion, brain TNF signaling pathway was activated by reperfusion, and inhibiting TNF-α with adalimumab significantly attenuated reperfusion-induced brain endothelial inflammation. In addition, the protein level of THBS1 was substantially upregulated upon reperfusion in brain endothelial cells and the peri-endothelial area in mice receiving cerebral ischemia.
Our study reveals the key molecular signatures of brain endothelial reperfusion injury and provides potential therapeutic targets for the treatment of brain microvascular injury after reperfusion therapy in ischemic stroke.
缺血性中风后的再灌注治疗常导致脑微血管损伤。然而，其潜在机制尚不清楚。
对氧糖剥夺（OGD）或OGD加恢复（OGD/R）后的人脑微血管内皮细胞进行转录组学和蛋白质组学分析，以鉴定因再灌注而失调的分子和信号通路。主要发现进一步在脑缺血再灌注小鼠模型中得到验证。
转录组学分析确定了OGD/R组和OGD组之间390个差异表达基因（DEG）。通路分析表明，这些基因大多与炎症相关，包括TNF信号通路、TGF-β信号通路、细胞因子-细胞因子受体相互作用、NOD样受体信号通路和NF-κB信号通路。蛋白质组学分析确定了201个差异表达蛋白（DEP），这些蛋白主要与细胞外基质破坏和重塑、内皮转运功能受损以及炎症反应有关。六个基因（DUSP1、JUNB、NFKBIA、NR4A1、SERPINE1和THBS1）在mRNA和蛋白质水平上均被OGD/R上调。在脑缺血再灌注小鼠中，再灌注激活了脑TNF信号通路，用阿达木单抗抑制TNF-α可显著减轻再灌注诱导的脑内皮炎症。此外，在接受脑缺血的小鼠的脑内皮细胞和内皮周围区域，再灌注后THBS1的蛋白水平显著上调。
我们的研究揭示了脑内皮再灌注损伤的关键分子特征，并为缺血性中风再灌注治疗后脑微血管损伤的治疗提供了潜在的治疗靶点。
