Xiamen Key Laboratory of Chiral Drugs, Medical College, Xiamen University, Xiamen, China; Department of Pharmacy, Xiamen Medical College, Xiamen, China.
Xiamen Key Laboratory of Chiral Drugs, Medical College, Xiamen University, Xiamen, China.
Brain Behav Immun. 2017 Nov;66:289-301. doi: 10.1016/j.bbi.2017.07.015. Epub 2017 Jul 21.
Propane-2-sulfonic acid octadec-9-enyl-amide (N15), an analogue of oleoylethanolamide (OEA), is a novel PPARα/γ dual agonist. Our previous studies verified the positive effects of OEA on the acute and delayed stages of cerebral ischemia. However, it is not clear whether N15 is effective against ischemic cerebral injury. In the present study, male Kunming mice were subjected to middle cerebral artery occlusion (MCAO). To evaluate its preventive effects, N15 (50, 100 or 200mg/kg, ip) was administered for 3days before ischemia. To evaluate its therapeutic effects, N15 (200mg/kg, ip) was administered 1h before reperfusion or 0, 1, 2 or 4h after reperfusion. Neurological deficit scores, infarct volume and the degree of brain oedema were determined at 24h after reperfusion. Blood brain barrier (BBB) disruption was evaluated by Evans blue (EB) and FITC-dextran leakages at 6h after reperfusion. The activation/inflammatory responses of microglia/macrophages were detected using immunohistochemistry and western blot. N15 pretreatment improved neurological dysfunction, reduced infarct volume and alleviated brain oedema in a dose-dependent manner; the most effective dose was 200mg/kg. The therapeutic time window was within 2h after reperfusion. N15 treatment preserved the BBB integrity and suppressed the activation of microglia/macrophages. N15 inhibited inflammatory cytokine expression not only in MCAO mice but also in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Additionally, N15 markedly decreased the phosphorylation levels of NF-κBp65, STAT3, and ERK1/2 both in vivo and in vitro. Furthermore, the PPARα antagonist MK886 or PPARγ antagonist T0070907 respectively partly abolished the anti-inflammatory effects of N15 in vitro. Our findings demonstrated that N15 can exert neuroprotective effects against cerebral ischemic insult. Moreover, the neuroprotective effects of N15 on cerebral ischemia may be attributed to its anti-inflammatory properties, at least in part, by enhancing PPARα/γ dual signaling and inhibiting the activation of the NF-κB, STAT3, and ERK1/2 signaling pathways. These findings suggest that N15 may be a potential therapeutic choice for the prevention and treatment of ischemic stroke.
丙烷-2-磺酸十八-9-烯基酰胺（N15）是一种新型的过氧化物酶体增殖物激活受体α/γ双重激动剂，是油酰乙醇酰胺（OEA）的类似物。我们之前的研究证实了 OEA 对急性和迟发性脑缺血的积极作用。然而，目前尚不清楚 N15 是否对缺血性脑损伤有效。在本研究中，雄性昆明小鼠接受大脑中动脉闭塞（MCAO）。为了评估其预防作用，在缺血前 3 天给予 N15（50、100 或 200mg/kg，ip）。为了评估其治疗作用，在再灌注前 1h 或再灌注后 0、1、2 或 4h 给予 N15（200mg/kg，ip）。在再灌注后 24h 测定神经功能缺损评分、梗死体积和脑水肿程度。在再灌注后 6h 通过 Evans 蓝（EB）和 FITC-葡聚糖渗漏评估血脑屏障（BBB）破坏。用免疫组织化学和 Western blot 检测小胶质细胞/巨噬细胞的激活/炎症反应。N15 预处理可改善神经功能障碍，减少梗死体积，减轻脑水肿，呈剂量依赖性；最有效剂量为 200mg/kg。治疗时间窗为再灌注后 2h 内。N15 治疗可维持 BBB 完整性并抑制小胶质细胞/巨噬细胞的激活。N15 不仅在 MCAO 小鼠中，而且在脂多糖（LPS）刺激的 BV-2 小胶质细胞中均抑制炎症细胞因子的表达。此外，N15 体内和体外均可显著降低 NF-κBp65、STAT3 和 ERK1/2 的磷酸化水平。此外，PPARα 拮抗剂 MK886 或 PPARγ 拮抗剂 T0070907 分别部分消除了 N15 在体外的抗炎作用。我们的研究结果表明，N15 可发挥神经保护作用，对抗脑缺血损伤。此外，N15 对脑缺血的神经保护作用可能归因于其抗炎特性，至少部分归因于增强 PPARα/γ 双重信号和抑制 NF-κB、STAT3 和 ERK1/2 信号通路的激活。这些发现表明，N15 可能是预防和治疗缺血性中风的潜在治疗选择。
