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 (10): 1734-1742.doi: 10.4103/1673-5374.257529

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

白藜芦醇对蛛网膜下腔出血的神经保护：抑制氧化应激、内质网应激和减轻神经炎症

  

  • 出版日期:2019-10-15 发布日期:2019-10-15
  • 基金资助:

    中国国家自然科学基金项目（81873768，81671213）山东省重点研究发展基金项目（2017GSF218091），山东大学基础研究基金（2015JC008）

Resveratrol reduces brain injury after subarachnoid hemorrhage by inhibiting oxidative stress and endoplasmic reticulum stress

Yun-Kai Xie 1, Xin Zhou 1, 2, Hong-Tao Yuan 1, Jie Qiu 1, Dan-Qing Xin 1, Xi-Li Chu 1, Da-Chuan Wang 1,  Zhen Wang 2   

  1. 1 Department of Physiology, School of Basic Medical Sciences, Shandong University, Jinan, Shandong Province, China
    2 Department of Spinal Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong Province, China
  • Online:2019-10-15 Published:2019-10-15
  • Contact: Da-Chuan Wang, PhD, wangdachuan@medmial.com.cn; Zhen Wang, PhD, wangzhen@sdu.edu.cn.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81873768 and 81671213 (both to ZW); the Key Research and Development Foundation of Shandong Province of China, No. 2017GSF218091 (to ZW); the Fundamental Research Funds of Shandong University of China, No. 2015JC008 (to ZW).

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摘要：

既往研究认为存在于多种植物中的生物活性物质——白藜芦醇可减少蛛网膜下腔出血后的早期脑损伤，但其具体机制尚不明确。实验通过小脑延髓池注射自体血建立实验性蛛网膜下腔出血大鼠模型，以探讨其机制。在损伤后2，6，24，46h腹腔注射60 mg/kg白藜芦醇进行治疗。损伤后48h以改良的Garcia评分评价神经功能，以干湿法测量脑水肿情况，以TUNEL染色检测前额皮质中神经细胞的凋亡，以比色法检测前额皮质中活性氧和丙二醛水平，以反转录PCR检测前额皮质中Chop，葡萄糖调节蛋白78、核转录因子E2相关因子2及血红素加氧酶1mRNA的表达水平，以ELISA检测前额皮质中肿瘤坏死因子α的含量，以免疫组化染色法检测核转录因子E2相关因子2、血红素加氧酶1、葡萄糖调节蛋白78、Chop以及胶质纤维酸性蛋白的阳性细胞数量，以Western blot分析大脑皮质中核转录因子E2相关因子2、血红素加氧酶1、葡萄糖调节蛋白78和Chop的蛋白表达水平。实验结果显示：（1）白藜芦醇可明显减轻实验性蛛网膜下腔出血大鼠模型的神经功能缺陷及脑水肿，并减少前额皮质中神经细胞的凋亡；（2）可降低前额皮质中活性氧和丙二醛的水平，上调核因子红细胞2相关因子2和血红素加氧酶1mRNA及蛋白的表达；（3）降低葡萄糖调节蛋白78和Chop mRNA及蛋白的表达以及肿瘤坏死因子α水平；（4）活化星形胶质细胞；（5）实验数据表明，白藜芦醇可通过减轻氧化损伤、内质网应激和神经炎症的机制实现对蛛网膜下腔出血的神经保护作用。

orcid: 0000-0002-7807-8938 (Da-Chuan Wang)

关键词: 白藜芦醇, 氧化应激, 内质网应激, 神经炎症, 蛛网膜下腔出血, 核转录因子E2相关因子2, 血红素加氧酶1, 葡萄糖调节蛋白78, 神经再生

Abstract:

Previous studies have shown that resveratrol, a bioactive substance found in many plants, can reduce early brain injury after subarach¬noid hemorrhage, but how it acts is still unclear. This study explored the mechanism using the experimental subarachnoid hemorrhage rat model established by injecting autologous blood into the cerebellomedullary cistern. Rat models were treated with an intraperitoneal injection of 60 mg/kg resveratrol 2, 6, 24 and 46 hours after injury. At 48 hours after injury, their neurological function was assessed using a modified Garcia score. Brain edema was measured by the wet-dry method. Neuronal apoptosis in the prefrontal cortex was detected by terminal deoxyribonucleotidyl transferase-mediated biotin-16-dUTP nick-end labeling assay. Levels of reactive oxygen species and malondialdehyde in the prefrontal cortex were determined by colorimetry. CHOP, glucose-regulated protein 78, nuclear factor-erythroid 2-related factor 2 and heme oxygenase-1 mRNA expression levels in the prefrontal cortex were measured by reverse transcription poly¬merase chain reaction. Tumor necrosis factor-alpha content in the prefrontal cortex was detected by enzyme linked immunosorbent assay. Immunohistochemical staining was used to detect the number of positive cells of nuclear factor-erythroid 2-related factor 2, heme oxygen¬ase 1, glucose-regulated protein 78, CHOP and glial fibrillary acidic protein. Western blot assay was utilized to analyze the expression levels of nuclear factor-erythroid 2-related factor 2, heme oxygenase 1, glucose-regulated protein 78 and CHOP protein expression levels in the prefrontal cortex. The results showed that resveratrol treatment markedly alleviated neurological deficits and brain edema in experimental subarachnoid hemorrhage rats, and reduced neuronal apoptosis in the prefrontal cortex. Resveratrol reduced the levels of reactive oxygen species and malondialdehyde, and increased the expression of nuclear factor-erythroid 2-related factor 2, heme oxygenase-1 mRNA and protein in the prefrontal cortex. Resveratrol decreased glucose-regulated protein 78, CHOP mRNA and protein expression and tumor necrosis factor-alpha level. It also activated astrocytes. The results suggest that resveratrol exerted neuroprotective effect on subarachnoid hemorrhage by reducing oxidative damage, endoplasmic reticulum stress and neuroinflammation. The study was approved by the Animals Ethics Committee of Shandong University, China on February 22, 2016 (approval No. LL-201602022).

Key words: nerve regeneration, resveratrol, oxidative stress, endoplasmic reticulum stress, neuroinflammation, subarachnoid hemorrhage, nuclear factor-erythroid 2-related factor 2, heme oxygenase-1, glucose-regulated protein 78, neural regeneration