Recombinant CCL17 Enhances Hematoma Resolution and Activation of CCR4/ERK/Nrf2/CD163 Signaling Pathway After Intracerebral Hemorrhage in Mice
Recombinant CCL17 Enhances Hematoma Resolution and Activation of CCR4/ERK/Nrf2/CD163 Signaling Pathway After Intracerebral Hemorrhage in Mice
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DOI：
10.1007/s13311-020-00908-4
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发表时间：
2020-08-11
影响因子：
5.7
通讯作者：
Tang, Jiping
中科院分区：
文献类型：
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作者：
Deng, Shuixiang;Sherchan, Prativa;Tang, Jiping
脑出血后血肿是导致预后不良的重要因素。促进小胶质细胞吞噬作用以增强血肿消退可能是ICH后恢复的重要治疗靶点。C-C趋化因子受体4（CCR 4）对于调节中枢神经系统中的免疫平衡是重要的。然而，CCR 4激活是否可以减轻ICH后的血肿仍然未知。我们的目的是评估CCL 17（CCR 4的特异性配体）治疗是否可以通过CCR 4/ERK/Nrf 2/CD 163途径促进ICH后血肿消退。共使用261只成年雄性CD 1小鼠。对小鼠进行纹状体内注射自体血液以诱导ICH，并随机分配接受重组CCL 17（rCCL 17）或在ICH后1小时鼻内施用的媒介物。为了阐明潜在的机制，在ICH诱导前1 h给予C 021（CCR 4和ML 385的选择性抑制剂）和Nrf 2的选择性抑制剂。在ICH前48 h通过脑室内注射给予CD 163的规则间隔短回文重复序列（CRISPR）敲除。进行脑水肿、短期和长期神经行为评价、血肿体积、血红蛋白含量、蛋白质印迹和免疫荧光染色。内源性CCL 17、CCR 4和CD 163表达增加，并在ICH后72 h达到峰值。CCR 4由小胶质细胞表达。与载体相比，用rCCL 17激活CCR 4显著改善神经行为评分并减少血肿体积和脑水肿。此外，rCCL 17处理显著促进ERK 1/2的磷酸化，增加Nrf 2的表达，并上调CD 163的表达。rCCL 17的保护作用通过施用C 021、ML 385和CD 163 CRISPR敲除而被消除。这项研究表明，在ICH后，使用rCCL 17激活CCR 4通过增加CD 163表达和CCR 4/ERK/Nrf 2通路激活促进血肿消退，从而减轻脑水肿并改善神经功能。总的来说，我们的研究表明，CCR 4激活可能是一种潜在的治疗策略，以减轻脑出血后早期脑损伤的血肿。
Hematoma is a crucial factor leading to poor prognosis after intracerebral hemorrhage (ICH). Promoting microglial phagocytosis to enhance hematoma resolution may be an important therapeutic target for recovery after ICH. C-C chemokine receptor 4 (CCR4) is important for regulating immune balance in the central nervous system. However, whether CCR4 activation can attenuate hematoma after ICH remains unknown. We aimed to evaluate whether CCL17 (a specific ligand of CCR4) treatment can promote hematoma resolution through CCR4/ERK/Nrf2/CD163 pathway after ICH. A total of 261 adult male CD1 mice were used. Mice were subjected to intrastriatal injection of autologous blood to induce ICH and randomly assigned to receive recombinant CCL17 (rCCL17) or vehicle which was administered intranasally at 1 h after ICH. To elucidate the underlying mechanism, C021, a selective inhibitor of CCR4 and ML385 and a selective inhibitor of Nrf2 were administered 1 h prior to ICH induction. Clustered regularly interspaced short palindromic repeats (CRISPR) knockout for CD163 was administered by intracerebroventricular injection at 48 h before ICH. Brain edema, short- and long-term neurobehavior evaluation, hematoma volume, hemoglobin content, western blot, and immunofluorescence staining were performed. Endogenous CCL17, CCR4, and CD163 expression increased and peaked at 72 h after ICH. CCR4 was expressed by microglia. CCR4 activation with rCCL17 significantly improved neurobehavioral scores and reduced hematoma volume and brain edema compared with vehicle. Moreover, rCCL17 treatment significantly promoted phosphorylation of ERK1/2, increased the expression Nrf2, and upregulated CD163 expression after ICH. The protective effects of rCCL17 were abolished by administration of C021, ML385, and CD163 CRISPR knockout. This study demonstrated that CCR4 activation with rCCL17 promoted hematoma resolution by increasing CD163 expression and CCR4/ERK/Nrf2 pathway activation after ICH, thereby reducing brain edema and improving neurological function. Overall, our study suggests that CCR4 activation may be a potential therapeutic strategy to attenuate hematoma in early brain injury after ICH.