Sun Zhao-Wei, Sun Zhao-Xin, Zhao Yun, Zhang Ling, Xie Fang, Wang Xue, Li Jin-Shan, Zhou Mao-Yang, Feng Hong, Qian Ling-Jia
Beijing Institute of Basic Medical Sciences, Beijing, 100850, China.
Tianjin Key Laboratory of Exercise Physiology and Sports Medicine, Institute of Sport, Exercise & Health, Tianjin University of Sport, Tianjin, 301617, China.
Fluids Barriers CNS. 2025 Apr 2;22(1):35. doi: 10.1186/s12987-025-00639-8.
Emerging evidence suggests that chronic stress compromises blood‒brain barrier (BBB) integrity by disrupting brain microvascular endothelial cells (BMECs), contributing to the development of cognitive impairments. Thus, targeting the BBB is expected to be a promising treatment strategy. The biological function of rutin has been investigated in neurological disorders; however, its regulatory role in stress-induced BBB damage and cognitive decline and the underlying mechanisms remain elusive.
In a chronic unpredictable mild stress (CUMS) mouse model, a fluorescent dye assay and behavioral tests, including a novel object recognition test and Morris water maze, were performed to evaluate the protective effects of rutin on BBB integrity and cognition. The effects of rutin on BMEC function were also investigated in hCMEC/D3 cells (a human brain microvascular endothelial cell line) in vitro. Furthermore, the molecular mechanisms by which rutin restores BBB endothelium dysfunction were explored via RNA-seq, quantitative real-time PCR, western blotting, immunofluorescence and chromatin immunoprecipitation. Finally, biotinylated tumor necrosis factor-α (TNF-α) was employed to test the influence of rutin on the ability of circulating TNF-α to cross the BBB.
We identified that rutin attenuated BBB hyperpermeability and cognitive impairment caused by the 8-week CUMS procedure. Moreover, rutin promoted the proliferation, migration and angiogenesis ability of BMECs, and the integrity of the cellular monolayer through positively regulating the expression of genes involved. Furthermore, rutin impeded histone deacetylase 1 (HDAC1) recruitment and stabilized H3K27ac to increase Claudin-5 protein levels. Ultimately, normalization of the hippocampal HDAC1‒Claudin-5 axis by rutin blocked the infiltration of circulating TNF-α into the brain parenchyma and alleviated neuroinflammation.
This work establishes a protective role of rutin in regulating BMEC function and BBB integrity, and reveals that rutin is a potential drug candidate for curing chronic stress-induced cognitive deficits.
新出现的证据表明，慢性应激通过破坏脑微血管内皮细胞（BMECs）损害血脑屏障（BBB）的完整性，从而导致认知障碍的发生。因此，针对血脑屏障有望成为一种有前景的治疗策略。芦丁的生物学功能已在神经疾病中得到研究；然而，其在应激诱导的血脑屏障损伤和认知衰退中的调节作用及其潜在机制仍不清楚。
在慢性不可预测轻度应激（CUMS）小鼠模型中，进行荧光染料检测和行为测试，包括新颖物体识别测试和莫里斯水迷宫，以评估芦丁对血脑屏障完整性和认知的保护作用。还在体外hCMEC/D3细胞（一种人脑微血管内皮细胞系）中研究了芦丁对BMEC功能的影响。此外，通过RNA测序、定量实时PCR、蛋白质印迹、免疫荧光和染色质免疫沉淀，探索了芦丁恢复血脑屏障内皮功能障碍的分子机制。最后，使用生物素化肿瘤坏死因子-α（TNF-α）来测试芦丁对循环TNF-α穿越血脑屏障能力的影响。
我们发现芦丁减轻了8周CUMS程序引起的血脑屏障高通透性和认知障碍。此外，芦丁通过正向调节相关基因的表达，促进了BMECs的增殖、迁移和血管生成能力以及细胞单层的完整性。此外，芦丁阻止组蛋白去乙酰化酶1（HDAC1）募集并稳定H3K27ac以增加Claudin-5蛋白水平。最终，芦丁使海马HDAC1-Claudin-5轴正常化，阻止循环TNF-α渗入脑实质并减轻神经炎症。
这项工作确立了芦丁在调节BMEC功能和血脑屏障完整性方面的保护作用，并揭示芦丁是治疗慢性应激诱导的认知缺陷的潜在候选药物。
