Receptor-interacting protein kinase 2 (RIPK2) profoundly contributes to post-stroke neuroinflammation and behavioral deficits with microglia as unique perpetrators.
Receptor-interacting protein kinase 2 (RIPK2) profoundly contributes to post-stroke neuroinflammation and behavioral deficits with microglia as unique perpetrators.
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受体相互作用蛋白激酶2(RIPK2)在卒中后神经炎症和行为缺陷中起重要作用，小胶质细胞是唯一的致病因子。
DOI：
10.1186/s12974-023-02907-6
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发表时间：
2023-09-30
影响因子：
9.3
通讯作者：
中科院分区：
文献类型：
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作者：
受体相互作用蛋白激酶2 （RIPK2）是一种丝氨酸/苏氨酸激酶，其活性通过与模式识别受体（PRRs）的关联以及随后的TAK1、NF-κB和MAPK通路激活来传播炎症信号。中风后，死亡和垂死细胞释放大量损伤相关分子模式（DAMPs），激活PRRs并启动强烈的炎症反应。我们假设RIPK2通过增强中风的神经炎症反应在中风损伤的进展中起破坏性作用，并且RIPK2的全局遗传缺失或小胶质细胞特异性条件缺失将在缺血性中风后起到保护作用。成年（3-6个月）雄性小鼠进行45分钟的短暂性大脑中动脉闭塞（tMCAO），然后进行24小时、48小时或28天的再灌注。老龄雄性和雌性小鼠（18-24月龄）永久性缺血性脑卒中48 h后处死。用TTC染色（24-48 h）或甲酚紫染色（28d）计算梗死体积。在指定时间点进行感觉运动测试（抓地力、垂直网格和开阔场地）。通过免疫印迹、ELISA、免疫组织化学和RT-qPCR评估血脑屏障（BBB）损伤、紧密连接蛋白、基质金属蛋白酶-9 （MMP-9）和神经炎症标志物。通过bulk RNA测序和nanoString®生成差异基因表达谱。与野生型对照组相比，Ripk2的全局基因缺失导致卒中后24小时梗死面积减小，神经炎症标志物减少。Ripk2全局缺失也改善了急性和长期行为结果，对卒中后28天减少梗死面积和死亡率有强大的影响。小胶质细胞Ripk2 （mKO）的条件缺失部分重现了我们在全球Ripk2缺陷小鼠中的结果，显示出在损伤后48小时内减少梗死体积和改善行为结果的作用。最后，大量转录组分析和nanoString数据表明，小胶质细胞中Ripk2缺失会降低与MAPK和NF-κB信号相关的基因，通过减少免疫细胞激活和外周免疫细胞侵袭来抑制脑卒中损伤后的神经炎症反应。这些结果揭示了RIPK2在缺血性脑卒中损伤的发病机制中迄今未知的作用，其中小胶质细胞起着独特的作用。本研究确定RIPK2是神经炎症信号的有效传播因子，突出了其作为卒中后干预治疗靶点的潜力。在线版本包含补充材料，可在10.1186/s12974-023-02907-6获得。
Receptor-interacting protein kinase 2 (RIPK2) is a serine/threonine kinase whose activity propagates inflammatory signaling through its association with pattern recognition receptors (PRRs) and subsequent TAK1, NF-κB, and MAPK pathway activation. After stroke, dead and dying cells release a host of damage-associated molecular patterns (DAMPs) that activate PRRs and initiate a robust inflammatory response. We hypothesize that RIPK2 plays a damaging role in the progression of stroke injury by enhancing the neuroinflammatory response to stroke and that global genetic deletion or microglia-specific conditional deletion of Ripk2 will be protective following ischemic stroke. Adult (3–6 months) male mice were subjected to 45 min of transient middle cerebral artery occlusion (tMCAO) followed by 24 h, 48 h, or 28 days of reperfusion. Aged male and female mice (18–24 months) were subjected to permanent ischemic stroke and sacrificed 48 h later. Infarct volumes were calculated using TTC staining (24–48 h) or Cresyl violet staining (28d). Sensorimotor tests (weight grip, vertical grid, and open field) were performed at indicated timepoints. Blood–brain barrier (BBB) damage, tight junction proteins, matrix metalloproteinase-9 (MMP-9), and neuroinflammatory markers were assessed via immunoblotting, ELISA, immunohistochemistry, and RT-qPCR. Differential gene expression profiles were generated through bulk RNA sequencing and nanoString®. Global genetic deletion of Ripk2 resulted in decreased infarct sizes and reduced neuroinflammatory markers 24 h after stroke compared to wild-type controls. Ripk2 global deletion also improved both acute and long-term behavioral outcomes with powerful effects on reducing infarct volume and mortality at 28d post-stroke. Conditional deletion of microglial Ripk2 (mKO) partially recapitulated our results in global Ripk2 deficient mice, showing reductive effects on infarct volume and improved behavioral outcomes within 48 h of injury. Finally, bulk transcriptomic profiling and nanoString data demonstrated that Ripk2 deficiency in microglia decreases genes associated with MAPK and NF-κB signaling, dampening the neuroinflammatory response after stroke injury by reducing immune cell activation and peripheral immune cell invasion. These results reveal a hitherto unknown role for RIPK2 in the pathogenesis of ischemic stroke injury, with microglia playing a distinct role. This study identifies RIPK2 as a potent propagator of neuroinflammatory signaling, highlighting its potential as a therapeutic target for post-stroke intervention. The online version contains supplementary material available at 10.1186/s12974-023-02907-6.
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影响因子：
5.3
作者：
He R;Yuan X;Lv X;Liu Q;Tao L;Meng J
通讯作者：
Meng J
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DOI：
10.1016/j.bbi.2020.02.001
发表时间：
2020-07
期刊：
Brain, behavior, and immunity
影响因子：
--
作者：
Ahnstedt H;Patrizz A;Chauhan A;Roy-O'Reilly M;Furr JW;Spychala MS;D'Aigle J;Blixt FW;Zhu L;Bravo Alegria J;McCullough LD
通讯作者：
McCullough LD
影响因子：
5.9
作者：
通讯作者：
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影响因子：
7.9
作者：
El-Hakim Y;Mani KK;Eldouh A;Pandey S;Grimaldo MT;Dabney A;Pilla R;Sohrabji F
通讯作者：
Sohrabji F
影响因子：
8.3
作者：
Bieber, Michael;Gronewold, Janine;Hermann, Dirk M.
通讯作者：
Hermann, Dirk M.