Cerebral cavernous malformation 3 relieves subarachnoid hemorrhage-induced neuroinflammation in rats through inhibiting NF-kB signaling pathway
Cerebral cavernous malformation 3 relieves subarachnoid hemorrhage-induced neuroinflammation in rats through inhibiting NF-kB signaling pathway
复制标题
脑海绵状血管瘤3通过抑制NF-kB信号通路减轻蛛网膜下腔出血所致大鼠神经炎症
DOI：
10.1016/j.brainresbull.2020.04.003
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发表时间：
2020-07-01
影响因子：
3.8
通讯作者：
Chen, Gang
中科院分区：
文献类型：
--
作者：
Peng, Wei;Wu, Xin;Chen, Gang
蛛网膜下腔出血（SAH）是一种严重的急性脑血管病，致残率和死亡率高。近年来，大量研究表明早期脑损伤（EBI）可能是SAH预后不良的重要原因，而小胶质细胞介导的神经炎症是EBI的重要病理过程。先前的研究表明，肿瘤坏死因子受体相关因子6（TRAF 6）参与SAH后小胶质细胞介导的神经炎症。此外，据报道，脑海绵状血管畸形3/哺乳动物不育20样激酶4（CCM 3/MST 4）直接磷酸化TRAF 6，以抑制其泛素化和限制炎症反应。然而，CCM 3/MST 4与SAH之间的关系尚未报道。本研究采用自体动脉血注入视交叉前池的方法建立了成年雄性大鼠蛛网膜下腔出血模型。此外，BV-2细胞，以及从用氧血红蛋白（OxyHb）处理24小时的大鼠的原代小胶质细胞培养物，被用作SAH的体外模型。采用免疫印迹、免疫荧光、Fluoro-JadeC染色、酶联免疫吸附试验（ELISA）和行为学测试等方法对实验动物进行了研究。脑组织中CCM 3/MST 4的表达无明显变化，而小胶质细胞中CCM 3的表达明显下降。我们还发现，在BV-2细胞中，CCM 3蛋白水平在OxyHb处理后下降，在处理后6 h达到最低点。相反，MST 4的蛋白水平没有显著变化。此外，我们概括了用OxyHb处理的原代小胶质细胞培养物的SAH体外模型中CCM 3表达的降低和CCM 3亚细胞定位的变化。CCM 3过表达可降低细胞变性、神经认知功能障碍、核内NF-κ B p65水平和炎症因子（TNF-α和IL-1 β）水平。这些结果表明CCM 3的过表达通过NF-κ B信号通路减轻脑损伤和神经损伤。
Subarachnoid hemorrhage (SAH) is a severe acute cerebrovascular disease with high rates of disability and death. In recent years, a large number of studies has shown that early brain injury (EBI) may be a crucial cause of the poor prognosis of SAH and that microglia-mediated neuroinflammation is an important pathological process in EBI. Previous studies have indicated that tumor necrosis factor receptor-associated factor 6 (TRAF6) is involved in microglia-mediated neuroinflammation after SAH. In addition, it has been reported that cerebral cavernous malformation 3/mammalian sterile20-like kinase 4 (CCM3/MST4) directly phosphorylates TRAF6 to inhibit its ubiquitination and to limit inflammatory responses. However, the association between CCM3/MST4 and SAH has not been reported. In our present study, we established a SAH model in adult male rats through injecting autologous arterial blood into the prechiasmatic cistern. Additionally, BV-2 cells, as well as primary microglial cultures from rats treated with oxygen hemoglobin (OxyHb) for 24 h, were used as in vitro models of SAH. Then, western blot, immunofluorescence, Fluoro-JadeC staining and Enzyme-linked immunosorbent assay (ELISA) and behavioral tests was applied in this study. We observed no significant change in the level of CCM3/MST4 in brain tissues, but a markedly decline of CCM3 in microglia of rats. We also found that the protein level of CCM3 was decreased in BV-2 cells after OxyHb treatment, reaching the lowest point at 6 h post-treatment. In contrast, there was no significant change in the protein level of MST4. Additionally, we recapitulated decreased expression of CCM3 and changes in subcellular localization of CCM3 in vitro model of SAH with primary microglial cultures treated with OxyHb. Overexpression of CCM3 decreased cellular degeneration, neurocognitive impairment, NF-kappa B p65 level in the nuclear, and inflammatory factors level (TNF-a and IL-1 beta). These results suggest that overexpression of CCM3 alleviated brain injury and neurological damage through the NF-kappa B signaling pathway.