Inhibiting NLRP3 inflammasome signaling pathway promotes neurological recovery following hypoxic-ischemic brain damage by increasing p97-mediated surface GluA1-containing AMPA receptors.
Inhibiting NLRP3 inflammasome signaling pathway promotes neurological recovery following hypoxic-ischemic brain damage by increasing p97-mediated surface GluA1-containing AMPA receptors.
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抑制NLRP 3炎性体信号通路通过增加p97介导的表面含GluA1的AMPA受体促进缺氧缺血性脑损伤后的神经恢复。
DOI：
10.1186/s12967-023-04452-5
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发表时间：
2023-08-24
影响因子：
7.4
通讯作者：
Dong, Zhifang
中科院分区：
文献类型：
--
作者：
Chen, Yuxin;Li, Xiaohuan;Xiong, Qian;Du, Yehong;Luo, Man;Yi, Lilin;Pang, Yayan;Shi, Xiuyu;Wang, Yu Tian;Dong, Zhifang
核苷酸结合的寡聚区(NOD)样受体蛋白3(NLRP3)炎症小体被认为是缺血性卒中所致神经炎症和继发性脑损伤的关键介质。然而，NLRP3炎症体在新生儿缺氧缺血性脑病(HIE)中的作用及其机制尚不清楚。免疫印迹法检测α-amino-3-hydroxy-5-methyl-4-isoxazole-propionicacid受体(AMPAR)亚单位NLRP3、半胱氨酸天冬氨酸特异性蛋白酶-1(Caspase-1)和白介素1-β(IL-1β)及含ATP酶Valosin蛋白(Vcp/p97)的蛋白表达。免疫共沉淀法测定p97与AMPA谷氨酸受体1(GluA1)的相互作用。用氯化三苯基四氮唑(TTC)染色观察缺氧缺血性脑损伤(HIBD)的组织病理学变化。用聚合酶链式反应(PCR)和Western blotting对基因敲除小鼠进行基因分型。运动功能，包括肌力和协调性，通过握力和旋转棒测试进行评估。用Morris水迷宫(MWM)测量大鼠的空间认知功能。我们报道了缺氧缺糖大鼠原代培养神经元中NLRP3炎症体信号通路，如NLRP3、Caspase-1和IL-1β被激活。进一步研究表明，HIBD大鼠海马区突触后膜AMPAR GluA1亚单位蛋白水平显著降低，经caspase-1抑制剂AC-YVAD-CMK治疗后可恢复至正常水平。同样，体外研究表明，OGD降低了培养的原代神经元质膜上的GluA1蛋白水平，而AC-YVAD-CMK处理则恢复了这种下降。重要的是，我们发现OGD处理明显增强了p97和GluA1之间的相互作用，而AC-YVAD-CMK处理促进了p97从GluA1复合体中解离，从而促进了GluA1在原代培养神经元质膜上的定位。最后，我们报道了HIBD动物在运动功能、学习和记忆方面的缺陷，可以通过药物干预或caspase-1的基因消融来改善。抑制NLRP3炎症体信号通路通过增加p97介导的表面GluA1表达促进HIBD动物神经功能的恢复，从而为HIE的治疗提供新的视角。网上版载有补充材料，可在10.1186/s12967-023-04452-5查阅。
The nucleotide-binding oligomeric domain (NOD)-like receptor protein 3 (NLRP3) inflammasome is believed to be a key mediator of neuroinflammation and subsequent secondary brain injury induced by ischemic stroke. However, the role and underlying mechanism of the NLRP3 inflammasome in neonates with hypoxic-ischemic encephalopathy (HIE) are still unclear. The protein expressions of the NLRP3 inflammasome including NLRP3, cysteinyl aspartate specific proteinase-1 (caspase-1) and interleukin-1β (IL-1β), the α-amino-3-hydroxy-5-methyl-4-isoxazole-propionicacid receptor (AMPAR) subunit, and the ATPase valosin-containing protein (VCP/p97), were determined by Western blotting. The interaction between p97 and AMPA glutamate receptor 1 (GluA1) was determined by co-immunoprecipitation. The histopathological level of hypoxic-ischemic brain damage (HIBD) was determined by triphenyltetrazolium chloride (TTC) staining. Polymerase chain reaction (PCR) and Western blotting were used to confirm the genotype of the knockout mice. Motor functions, including myodynamia and coordination, were evaluated by using grasping and rotarod tests. Hippocampus-dependent spatial cognitive function was measured by using the Morris-water maze (MWM). We reported that the NLRP3 inflammasome signaling pathway, such as NLRP3, caspase-1 and IL-1β, was activated in rats with HIBD and oxygen-glucose deprivation (OGD)-treated cultured primary neurons. Further studies showed that the protein level of the AMPAR GluA1 subunit on the hippocampal postsynaptic membrane was significantly decreased in rats with HIBD, and it could be restored to control levels after treatment with the specific caspase-1 inhibitor AC-YVAD-CMK. Similarly, in vitro studies showed that OGD reduced GluA1 protein levels on the plasma membrane in cultured primary neurons, whereas AC-YVAD-CMK treatment restored this reduction. Importantly, we showed that OGD treatment obviously enhanced the interaction between p97 and GluA1, while AC-YVAD-CMK treatment promoted the dissociation of p97 from the GluA1 complex and consequently facilitated the localization of GluA1 on the plasma membrane of cultured primary neurons. Finally, we reported that the deficits in motor function, learning and memory in animals with HIBD, were ameliorated by pharmacological intervention or genetic ablation of caspase-1. Inhibiting the NLRP3 inflammasome signaling pathway promotes neurological recovery in animals with HIBD by increasing p97-mediated surface GluA1 expression, thereby providing new insight into HIE therapy. The online version contains supplementary material available at 10.1186/s12967-023-04452-5.
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DOI：
10.1074/jbc.m901203200
发表时间：
2009-05-22
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dixon RM;Mellor JR;Hanley JG
通讯作者：
Hanley JG
影响因子：
6.1
作者：
Gerace, E.;Masi, A.;Moroni, F.
通讯作者：
Moroni, F.
影响因子：
16.2
作者：
Diering GH;Huganir RL
通讯作者：
Huganir RL
影响因子：
5.1
作者：
Fann, David Yang-Wei;Lim, Yun-An;Arumugam, Thiruma V.
通讯作者：
Arumugam, Thiruma V.
影响因子：
15.9
作者：
Gunn, AJ;Gunn, TR;Gluckman, PD
通讯作者：
Gluckman, PD