中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (4): 605-612.doi: 10.4103/1673-5374.247465

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

刺槐素通过NLRP3信号通路对抗脑缺血再灌注

  

  • 出版日期:2019-04-15 发布日期:2019-04-15
  • 基金资助:

    新疆维吾尔自治区自然科学基金（2016D01C120）

Acacetin protects against cerebral ischemia-reperfusion injury via the NLRP3 signaling pathway

Juan Bu 1 , Shen Shi 2 , Hui-Qin Wang 1 , Xiao-Shan Niu 3 , Zong-Feng Zhao 1 , Wei-Dong Wu 1 , Xiao-Ling Zhang 1 , Zhi Ma 3 , Yan-Jun Zhang 1 , Hui Zhang 1 , Yi Zhu 3   

  1. 1 Clinical Research Center, People’s Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang Uygur Autonomous Region, China
    2 Laboratory Animal Research Center, Center for Disease Control and Prevention, Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang Uygur Autonomous Region, China
    3 Department of Neurology, People’s Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang Uygur Autonomous Region, China
  • Online:2019-04-15 Published:2019-04-15
  • Contact: Yi Zhu, MS, zhuyixinjiang@sina.com.
  • Supported by:

    This study was supported by the Natural Science Foundation of Xinjiang Uygur Autonomous Region of China, No. 2016D01C120 (to JB).

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摘要：

刺槐素是一种潜在的神经保护剂，对LPS诱导的神经炎性反应有抑制作用，但其对脑缺血再灌注损伤后炎症小体3(NLRP3)是否有影响尚少见报道。为此，实验设计用线栓法栓塞C57BL/6小鼠，建立局灶性脑缺血再灌注损伤模型，于再灌注时(缺血1h)腹腔注射25 mg/kg刺槐素设为刺槐素组，将再灌注时腹腔注射等体积(0.1 mL/10 g)生理盐水设为缺血再灌注组。为了解刺槐素对脑缺血再灌注损伤的作用，采用2，3，5-氯化三苯基四氮唑染色法和Zea-Longa评分法评估比较，刺槐素组神经功能评分和脑梗死体积均显著降低。为了解刺槐素对脑缺血再灌注损伤后小胶质细胞介导的炎症反应的影响，采用免疫组织染色法测定小鼠缺血侧脑组织海马区小胶质细胞标记物钙接头蛋白(Ibal)的表达，酶联免疫法检测了小鼠缺血侧脑组织肿瘤坏死因子α，白细胞介素1β，白细胞介素6蛋白的表达，发现刺槐素组缺血侧脑组织海马区Ibal蛋白表达，缺血侧脑组织肿瘤坏死因子α，白细胞介素1β和白细胞介素6蛋白表达表达显著低于缺血再灌注组。蛋白印迹法检测结果显示，刺槐素组缺血侧脑组织Toll样受体4(TLR4)，核转录因κB(NF-kB)，NLRP3，半胱氨酸蛋白酶1前体，半胱氨酸蛋白酶1，白细胞介素1β前体和白细胞介素1β蛋白表达均显著低于缺血再灌注组。上述结果表明，刺槐素对脑缺血再灌注损伤具有保护作用，其作用机制与抑制小胶质细胞介导的炎症反应，进而抑制TLR4/NF-kB/NLRP3信号通路相关。

orcid: 0000-0002-1783-7101(Yi Zhu)

关键词: 刺槐素, 脑缺血再灌注, 小胶质细胞, 神经保护, 炎症小体3, 炎性因子, 小鼠, 梗死体积, 信号通路, 核转录因子kB, 神经再生

Abstract:

Acacetin (5,7-dihydroxy-4′-methoxyflavone), a potential neuroprotective agent, has an inhibitory effect on lipopolysaccharide-in¬duced neuroinflammatory reactions. However, whether acacetin has an effect on inflammatory corpuscle 3 (NLRP3) after cerebral ischemia-reperfusion injury has not been fully determined. This study used an improved suture method to establish a cerebral isch¬emia-reperfusion injury model in C57BL/6 mice. After ischemia with middle cerebral artery occlusion for 1 hour, reperfusion with intraperitoneal injection of 25 mg/kg of acacetin (acacetin group) or an equal volume of saline (0.1 mL/10 g, middle cerebral artery occlu¬sion group) was used to investigate the effect of acacetin on cerebral ischemia-reperfusion injury. Infarct volume and neurological function scores were determined by 2,3,5-triphenyltetrazolium chloride staining and the Zea-Longa scoring method. Compared with the middle cerebral artery occlusion group, neurological function scores and cerebral infarction volumes were significantly reduced in the acacetin group. To understand the effect of acacetin on microglia-mediated inflammatory response after cerebral ischemia-reperfusion injury, im¬munohistochemistry for the microglia marker calcium adapter protein ionized calcium-binding adaptor molecule 1 (Iba1) was examined in the hippocampus of ischemic brain tissue. In addition, tumor necrosis factor-α, interleukin-1β, and interleukin-6 expression in ischemic brain tissue of mice was quantified by enzyme-linked immunosorbent assay. Expression of Iba1, tumor necrosis factor-α, interleukin-1β and interleukin-6 was significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Western blot assay results showed that expression of Toll-like receptor 4, nuclear factor kappa B, NLRP3, procaspase-1, caspase-1, pro-interleukin-1β, and interleukin-1β were significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Our findings indicate that acacetin has a protective effect on cerebral ischemia-reperfusion injury, and its mechanism of action is associated with inhibi¬tion of microglia-mediated inflammation and the NLRP3 signaling pathway.

Key words: nerve regeneration, acacetin, cerebral ischemia-reperfusion injury, microglia, NLRP3 inflammasome, inflammatory factor, infarct volume, signaling pathway, nuclear factor-κB, neuroprotection, neural regeneration