Background: Stroke is the third leading cause of disability and mortality in the world. The c omplement C3a receptor plays a prominent role in post stroke brain inflammation. We and others have reported that either genetic deficiency of complement C3a receptor (C3aR) or its pharmacological inhibition in rodents protect against cerebral ischemia. The existing C3aR antagonist (SB290157) is limited by its reported agonist effect in different model systems. Hypothesis: The novel highly-selective C3aR antagonist JR14a confers robust neuroprotection in stroke Methods: Mouse primary brain endothelial cells (cell biologics Inc. IL, USA) were seeded at a density of 5000 cells/well in 96-well plates and cultured until 80-90% confluency. Endothelial cytotoxicity was tested using an LDH assay following 24 hours of exposure to JR14a. Oxygen-glucose deprivation (OGD) was followed by reperfusion to compare the in-vitro effects of JR14a and SB290157, using ELISA performed for TNF-α and IL-6 and immunofluorescence for ICAM-1 expression. We also compared JR14a and SB290157 in the middle cerebral artery occlusion (MCAO, C57BL/6) stroke model, evaluating infarct volume. Results: JR14a treated endothelial cells exhibited reduced LDH release (Fig.1) and more potent anti-inflammatory response following OGD/R compared to SB290157. We found that JR14a treatment significantly reduced endothelial TNF-α, IL-6 and ICAM-1 expression. JR14a also significantly reduced brain infarction and microglial activation relative to SB290157 following MCAO in C57BL/6 mice (Fig 2). Conclusion: We conclude that the novel C3aR inhibitor JR14a attenuates inflammation and endothelial dysfunction in stroke and may confer a more potent neuroprotective effect than SB290157. Further work is warranted to explore the mechanisms underlying JR14a-mediated neuroprotection

中文翻译：

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摘要 15：新型补体 C3ar 拮抗剂 Jr14a 在中风中具有神经保护作用

背景：中风是世界上导致残疾和死亡的第三大原因。补体 C3a 受体在中风后脑部炎症中起重要作用。我们和其他人报道了补体 C3a 受体 (C3aR) 的遗传缺陷或其在啮齿动物中的药理学抑制作用可以防止脑缺血。现有的 C3aR 拮抗剂 (SB290157) 受到其在不同模型系统中报告的激动剂作用的限制。假设：新型高选择性 C3aR 拮抗剂 JR14a 在中风中具有强大的神经保护作用 方法：将小鼠原代脑内皮细胞（cell biologics Inc. IL，USA）以 5000 个细胞/孔的密度接种在 96 孔板中并培养至 80 -90% 汇合。在暴露于 JR14a 24 小时后，使用 LDH 测定法测试内皮细胞毒性。氧葡萄糖剥夺 (OGD) 后再灌注以比较 JR14a 和 SB290157 的体外效应，使用 ELISA 对 TNF-α 和 IL-6 进行，免疫荧光对 ICAM-1 表达进行比较。我们还在大脑中动脉闭塞 (MCAO, C57BL/6) 中风模型中比较了 JR14a 和 SB290157，评估了梗死体积。结果：与 SB290157 相比，经过 JR14a 处理的内皮细胞在 OGD/R 后表现出 LDH 释放减少（图 1）和更有效的抗炎反应。我们发现 JR14a 治疗显着降低了内皮 TNF-α、IL-6 和 ICAM-1 的表达。相对于 SB290157，在 C57BL/6 小鼠的 MCAO 之后，JR14a 还显着降低了脑梗塞和小胶质细胞活化（图 2）。结论：我们得出结论，新型 C3aR 抑制剂 JR14a 可减轻中风的炎症和内皮功能障碍，并且可能比 SB290157 具有更有效的神经保护作用。需要进一步的工作来探索 JR14a 介导的神经保护机制