Department of Neurosurgery, West China Medical School, West China Hospital, Sichuan University, Chengdu, China.
Department of Neurosurgery, Chengdu Second People's hospital, Chengdu, China.
Cell Prolif. 2021 Sep;54(9):e13108. doi: 10.1111/cpr.13108. Epub 2021 Aug 9.
Necroptosis is widespread in neurodegenerative diseases. Here, we examined necroptosis in the hippocampus and cortex after hydrocephalus and found that a necroptosis pathway inhibitor alleviates necroptosis and provides neuroprotective effects.
Hydrocephalus was induced in C57BL/6 mice by kaolin. Haematoxylin and eosin (HE), Nissl, PI and Fluoro-Jade B (FJB) staining were used for general observations. Phosphorylated receptor-interacting protein kinase 3 (p-RIP3) and phosphorylated mixed lineage kinase domain-like (p-MLKL) were measured by Western blotting and immunohistochemistry. Scanning electron microscopy (SEM) was used to observe ependymal cilia. Magnetic resonance imaging (MRI) and the Morris water maze (MWM) test were used to assess neurobehavioral changes. Immunofluorescence was used to detect microglial and astrocyte activation. Inflammatory cytokines were measured by Western blotting and RT-PCR.
Obvious pathological changes appeared in the hippocampus and cortex after hydrocephalus, and expression of the necroptosis markers p-RIP3, p-MLKL and inflammatory cytokines increased. Necrostatin-1 (Nec-1) and GSK872 reduced necrotic cell death, attenuated p-RIP3 and p-MLKL levels, slightly improved neurobehaviours and inhibited microglial and astrocyte activation and inflammation.
RIP1/RIP3/MLKL mediates necroptosis in the cortex and hippocampus in a hydrocephalus mouse model, and Nec-1 and GSK872 have some neuroprotective effects.
细胞坏死性凋亡广泛存在于神经退行性疾病中。在此，我们研究了脑积水后海马体和皮质中的细胞坏死性凋亡，并发现细胞坏死性凋亡途径抑制剂可减轻细胞坏死性凋亡并提供神经保护作用。
通过高岭土诱导 C57BL/6 小鼠发生脑积水。苏木精和伊红（HE）、尼氏染色、碘化丙啶（PI）和氟硼二吡咯（FJB）染色用于一般观察。通过 Western blot 和免疫组织化学检测磷酸化受体相互作用蛋白激酶 3（p-RIP3）和磷酸化混合谱系激酶结构域样蛋白（p-MLKL）。扫描电子显微镜（SEM）用于观察室管膜纤毛。磁共振成像（MRI）和 Morris 水迷宫（MWM）测试用于评估神经行为变化。免疫荧光用于检测小胶质细胞和星形胶质细胞的激活。通过 Western blot 和 RT-PCR 测量炎性细胞因子。
脑积水后海马体和皮质出现明显的病理变化，细胞坏死性凋亡标志物 p-RIP3、p-MLKL 和炎性细胞因子表达增加。Necrostatin-1（Nec-1）和 GSK872 减少坏死性细胞死亡，降低 p-RIP3 和 p-MLKL 水平，轻度改善神经行为，并抑制小胶质细胞和星形胶质细胞激活和炎症。
RIP1/RIP3/MLKL 在脑积水小鼠模型中介导皮质和海马体的细胞坏死性凋亡，Nec-1 和 GSK872 具有一定的神经保护作用。
