Du Shibin, Deng Youliang, Yuan Hongjie, Sun Yanyan
Department of Anesthesiology, Shenzhen University General Hospital, Shenzhen University Clinical Medical Academy, Shenzhen University, Shenzhen 518055, China.
Department of Anesthesiology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
Evid Based Complement Alternat Med. 2019 Feb 3;2019:7219740. doi: 10.1155/2019/7219740. eCollection 2019.
Inflammation had showed its important role in the pathogenesis of cerebral ischemia and secondary damage. Safflower yellow B (SYB) had neuroprotective effects against oxidative stress-induced brain injuries, but the mechanisms were still largely unknown to us. In this study, we tried to investigate the anti-inflammation effects of SYB and the possible roles of AMPK/NF-B signaling pathway on these protective effects. In vivo, brain ischemia/reperfusion (I/R) was induced by transient middle cerebral artery occlusion for 2 h and reperfusion for 20 h. Neurofunctional evaluation, infarction area, and brain water contents were measured. Brain injury markers and inflammatory cytokines levels were measured by ELISA kits. In vitro, cell viability, apoptosis, and LDH leakage were measured after I/R in PC12 cells. The expression and phosphorylation levels of AMPK, NF-B p65, and P-IB- in cytoplasm and nuclear were measured by Western blotting. SiRNA experiment was performed to certify the role of AMPK. The results showed SYB reduced infarct size, improved neurological outcomes, and inhibited brain injury after I/R. test, SYB treatment alleviated PC12 cells injury and apoptosis and inhibited the inflammatory cytokines (IL-1, IL-6, TNF-, and COX-2) in a dose-dependent manner. SYB treatment induced AMPK phosphorylation and inhibited NF-B p65 nuclear translocation both in brain and in PC12 cells. Further studies also showed that the inhibition of NF-B activity of SYB was through AMPK. In conclusion, SYB protected brain I/R injury through reducing expression of inflammatory cytokines and this effect might be partly due to the inhibition of NF-B mediated by AMPK.
炎症在脑缺血及其继发性损伤的发病机制中发挥着重要作用。红花黄色素B（SYB）对氧化应激诱导的脑损伤具有神经保护作用，但其机制我们仍知之甚少。在本研究中，我们试图探究SYB的抗炎作用以及AMPK/NF-κB信号通路在这些保护作用中可能发挥的作用。在体内，通过短暂阻断大脑中动脉2小时并再灌注20小时诱导脑缺血/再灌注（I/R）。进行神经功能评估、测量梗死面积和脑含水量。使用ELISA试剂盒检测脑损伤标志物和炎性细胞因子水平。在体外，对PC12细胞进行I/R处理后，检测细胞活力、凋亡情况以及乳酸脱氢酶（LDH）泄漏。通过蛋白质免疫印迹法检测细胞质和细胞核中AMPK、NF-κB p65以及P-IκB-α的表达和磷酸化水平。进行小干扰RNA（SiRNA）实验以证实AMPK的作用。结果显示，SYB可减小梗死面积，改善神经功能结局，并抑制I/R后的脑损伤。检测发现，SYB处理可减轻PC12细胞损伤和凋亡，并以剂量依赖性方式抑制炎性细胞因子（IL-1、IL-6、TNF-α和COX-2）。SYB处理可诱导大脑和PC12细胞中AMPK磷酸化并抑制NF-κB p65核转位。进一步研究还表明，SYB对NF-κB活性的抑制作用是通过AMPK实现的。总之，SYB通过降低炎性细胞因子的表达来保护脑I/R损伤，且这种作用可能部分归因于AMPK介导的对NF-κB的抑制。
