Inhibition of GSK3β and RIP1K Attenuates Glial Scar Formation Induced by Ischemic Strokevia Reduction of Inflammatory Cytokine Production
Inhibition of GSK3β and RIP1K Attenuates Glial Scar Formation Induced by Ischemic Strokevia Reduction of Inflammatory Cytokine Production
复制标题
抑制 GSK3 β 和 RIP1K 可减轻缺血性中风引起的神经胶质疤痕形成，减少炎症细胞因子的产生
DOI：
10.3389/fphar.2020.00812
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发表时间：
2020-06-12
影响因子：
5.6
通讯作者：
Zhang, Hui-Ling
中科院分区：
文献类型：
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作者：
Liu, Jin;Zhu, Yong-Ming;Zhang, Hui-Ling
在缺血性脑卒中后的慢性期，胶质瘢痕可以阻止轴突再生和炎症的加剧。抑制糖原合成酶激酶-3 β (GSK3 β)或受体相互作用蛋白1激酶（RIP1K）对缺血性卒中的保护作用已有报道。目前的研究检测了RIP1K和GSK3 β在缺血性卒中诱导的胶质瘢痕形成中的作用。为了研究这一点，我们使用了基于大脑中动脉闭塞90分钟再灌注7天的缺血性卒中的动物模型，以及在原代培养的星形胶质细胞中进行缺氧和葡萄糖剥夺6小时再氧合24小时的动物模型。在体内和体外，我们发现SB216763 （GSK3 β抑制剂）和nec1 （RIP1K抑制剂）降低了胶质瘢痕标志物的水平，包括胶质纤维酸性蛋白（GFAP），神经蛋白和磷蛋白。SB216763和Nec-1还能降低炎症相关细胞因子的水平，包括白细胞介素-6 （IL-6）、白细胞介素-1 β (IL-1 β)和肿瘤坏死因子- α (tnf - α)。而只有Nec-1增加了白细胞介素-1受体拮抗剂的水平。与单独使用每种抗体相比，同时中和tnf - α、IL-1 β和IL-6的抗体可以更好地减少氧和葡萄糖剥夺引起的疤痕标记物的增加。进一步的研究表明，SB216763降低了坏死相关蛋白的水平，包括RIP1K、p-RIP1K、RIP3K、p-RIP3K、混合谱系激酶结构域样蛋白（MLKL）和p-MLKL，而Nec-1降低了p-GSK3 β的表达。与单独使用Nec-1 （10 μ M）和SB216763 （1 μ M）相比，Nec-1和SB216763联合使用可降低GFAP、神经can和炎症相关细胞因子的水平。综上所述，抑制GSK3 β或RIP1K可减少缺血性卒中诱导的胶质瘢痕形成。其潜在机制可能至少部分与降低炎症相关细胞因子水平和阻断GSK3 β -和ripk1介导途径之间的相互作用有关。
In the chronic phase following ischemic stroke, glial scars can prevent axonal regeneration and the intensification of inflammation. The protective effect of inhibition of glycogen synthase kinase-3 beta (GSK3 beta) or receptor-interacting protein 1 kinase (RIP1K) on ischemic stroke has been previously reported. The current study examined the effects of RIP1K and GSK3 beta on ischemic stroke-induced glial scar formation. To investigate this, we used anin vivomodel of ischemic stroke based on middle cerebral artery occlusion for 90 min followed by reperfusion for 7 d, and anin vitromodel in primary cultured astrocytes involving oxygen and glucose deprivation for 6 h followed by reoxygenation for 24 h. Bothin vivoandin vitro, we found that SB216763, a GSK3 beta inhibitor, and necrostatin-1 (Nec-1), a RIP1K inhibitor, decreased levels of glial scar markers, including glial fibrillary acidic protein (GFAP), neurocan, and phosphacan. SB216763 and Nec-1 also decreased levels of inflammatory related cytokines, including interleukin-6 (IL-6), interleukin-1 beta (IL-1 beta), and tumor necrosis factor-alpha (TNF-alpha). However, only Nec-1 increased the level of interleukin-1 receptor antagonist. Concurrent neutralization of TNF-alpha, IL-1 beta, and IL-6 with their antibodies provided better reduction in oxygen and glucose deprivation-induced increases in scar markers than obtained with separate use of each antibody. Further investigations showed that SB216763 reduced the levels of necroptosis-related proteins, including RIP1K, p-RIP1K, RIP3K, p-RIP3K, mixed lineage kinase domain-like protein (MLKL), and p-MLKL, while Nec-1 decreased the expression of p-GSK3 beta. Compared with Nec-1 (10 mu M) and SB216763 (1 mu M) alone, Nec-1 and SB216763 in combination reduced levels of GFAP, neurocan, and inflammatory-related cytokines. In conclusion, inhibition of GSK3 beta or RIP1K reduced glial scar formation induced by ischemic stroke. The underlying mechanisms might be at least, partially related to reducing levels of inflammatory-related cytokines and to blocking an interaction between GSK3 beta- and RIP1K-mediated pathways.