南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (2): 323-330.doi: 10.12122/j.issn.1673-4254.2023.02.23

• • 上一篇    

大豆异黄酮可减轻大鼠脑缺血/再灌注损伤：基于抑制铁死亡及炎症级联反应

李 晒，李 丽，闵思敏，刘赛赛，秦志文，熊志尚，徐建国，王博文，丁渡山，赵士弟   

  1. 蚌埠医学院临床医学院，病理生理学教研室，心脑血管疾病基础与临床重点实验室，安徽 蚌埠 233000
  • 出版日期:2023-02-20 发布日期:2023-03-16

Soybean isoflavones alleviate cerebral ischemia/reperfusion injury in rats by inhibiting ferroptosis and inflammatory cascade reaction

LI Shai, LI Li, MIN Simin, LIU Saisai, QIN Zhiwen, XIONG Zhishang, XU Jianguo, WANG Bowen, DING Dushan, ZHAO Shidi   

  1. Department of Clinical Medicine, Department of Pathophysiology, Key Laboratory of Basic and Clinical Cardiovascular and Cerebrovascular Diseases, Bengbu Medical College, Bengbu 233000, China
  • Online:2023-02-20 Published:2023-03-16

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摘要： 目的 基于大豆异黄酮（SI）对区域脑血流（rCBF）、铁死亡、炎症反应和血脑屏障（BBB）通透性的级联调控作用，探讨SI抗脑缺血/再灌注损伤机制。方法 将120只雄性SD大鼠随机分为假手术组（Sham组）、缺血/再灌注组（I/R组）和大豆异黄酮预处理组（SI组），40只/组。采用改良线栓法合并激光多普勒血流仪（LDF）建立大鼠局灶性脑缺血/再灌注损伤模型。术中各组大鼠通过LDF实时监测脑皮层血流动态变化；术后通过光镜和透射电镜观察脑组织病理形态、神经元和BBB超微结构变化，评估神经缺损程度，量化脑梗死、脑水肿和BBB破坏程度，分光光度法检测缺血半暗带Fe2+、谷胱甘肽（GSH）、丙二醛（MDA）、髓过氧化物酶（MPO）含量，ELISA法测定血清TNF-α和IL-1β活性，Western blot法和IHC法检测病灶周围GPX4、MMP-9和Occludin表达。结果 插线成功后I/R组和SI组大鼠rCBF均呈现骤降趋势，而拔线后I/R组rCBF上升慢于SI组；rCBF存在插线后不稳定和拔线后未复流状况。I/R组神经细胞固缩严重，线粒体变小、膜密度增加、嵴减少，紧密连接松散、基底膜厚薄不均，SI组各结构损伤相较于I/R组呈不同程度减轻。与Sham组相比，I/R组神经缺损评分、脑梗死体积、脑含水量和伊文思蓝渗透量明显增加（P<0.01），Fe2+和MDA含量增多、GSH含量及GPX4表达降低（P<0.01），MPO含量和血清TNF-α、IL-1β活性增高（P<0.01），MMP-9表达增多、Occludin表达减少（P<0.01）；与I/R组相比，SI组神经缺损评分、脑梗死体积、脑含水量和伊文思蓝渗透量降低（P<0.05或P<0.01），Fe2+和MDA含量减少、GSH含量及GPX4表达升高（P<0.05或P<0.01），MPO含量和血清TNF-α、IL-1β活性降低（P<0.05或P<0.01），MMP-9表达下调、Occludin表达上调（P<0.05，P<0.01）。结论 SI预处理可能通过改善脑血流，抑制铁死亡、炎症反应和保护BBB，进而减轻脑缺血/再灌注损伤。

关键词: 大豆异黄酮；脑缺血/再灌注；rCBF；铁死亡；炎症反应；血脑屏障

Abstract: Objective To explore the mechanism that mediates the effect of soybean isoflavones (SI) against cerebral ischemia/reperfusion (I/R) injury in light of the regulation of regional cerebral blood flow (rCBF), ferroptosis, inflammatory response and blood-brain barrier (BBB) permeability. Methods A total of 120 male SD rats were equally randomized into sham-operated group (Sham group), cerebral I/R injury group and SI pretreatment group (SI group). Focal cerebral I/R injury was induced in the latter two groups using a modified monofilament occlusion technique, and the intraoperative changes of real-time cerebral cortex blood flow were monitored using a laser Doppler flowmeter (LDF). The postoperative changes of cerebral pathological morphology and the ultrastructure of the neurons and the BBB were observed with optical and transmission electron microscopy. The neurological deficits of the rats was assessed, and the severities of cerebral infarction, brain edema and BBB disruption were quantified. The contents of Fe2 + , GSH, MDA and MPO in the ischemic penumbra were determined with spectrophotometric tests. Serum levels of TNF-α and IL-1βwere analyzed using ELISA, and the expressions of GPX4, MMP-9 and occludin around the lesion were detected with Western blotting and immunohistochemistry. Results The rCBF was sharply reduced in the rats in I/R group and SI group after successful insertion of the monofilament. Compared with those in Sham group, the rats in I/R group showed significantly increased neurological deficit scores, cerebral infarction volume, brain water content and Evans blue permeability (P<0.01), decreased Fe2 + level, increased MDA level, decreased GSH content and GPX4 expression (P<0.01), increased MPO content and serum levels of TNF-α and IL-1β (P<0.01), increased MMP-9 expression and lowered occludin expression (P<0.01). All these changes were significantly ameliorated in rats pretreated with IS prior to I/R injury (P<0.05 or 0.01). Conclusion SI preconditioning reduces cerebral I/R injury in rats possibly by improving rCBF, inhibiting ferroptosis and inflammatory response and protecting the BBB.

Key words: soybean isoflavones; cerebral ischemia/reperfusion; regional cerebral blood flow; ferroptosis; inflammatory response; blood-brain barrier