南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (12): 2690-2698.doi: 10.12122/j.issn.1673-4254.2025.12.16

• • 上一篇    

白藜芦醇改善PM2.5诱导的脑缺血再灌注损伤小鼠血脑屏障及维持线粒体动力学平衡

秦萌1,2(), 孙思宇2,3, 刘佳琪2, 高玉娇1,2, 汪昊2,3, 王友坤2, 孙奥2, 严加纯2, 汪金宝2, 于影1,2()   

  1. 1.蚌埠医科大学,生理学教研室,安徽 蚌埠 233000
    2.蚌埠医科大学,心脑血管疾病基础与临床重点实验室,安徽 蚌埠 233000
    3.蚌埠医科大学,流行病与卫生统计学教研室,安徽 蚌埠 233000
  • 收稿日期:2025-04-14 出版日期:2025-12-20 发布日期:2025-12-22
  • 通讯作者: 于影 E-mail:13017518983@163.com;yuying2011@126.com
  • 作者简介:秦 萌,在读硕士研究生,E-mail:13017518983@163.com
  • 基金资助:
    安徽省自然科学基金(2108085MH252);蚌埠医学院512人才培育计划项目(by51201307);蚌埠医科大学研究生科研创新计划项目(Byycx24016);蚌埠医科大学研究生科研创新计划项目(Byycx24117);国家级大学生创新训练项目(2023103670013)

Resveratrol protects barrier function of mouse brain microvascular endothelial cell monolayers with oxygen/glucose deprivation and PM2.5 exposure by maintaining mitochondrial dynamics balance

Meng QIN1,2(), Siyu SUN2,3, Jiaqi LIU2, Yujiao GAO1,2, Hao WANG2,3, Youkun WANG2, Ao SUN2, Jiachun YAN2, Jinbao WANG2, Ying YU1,2()   

  1. 1.Department of Physiology, Bengbu Medical University, Bengbu 233000, China
    2.Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical University, Bengbu 233000, China
    3.Department of Epidemiology and Health Statistics, Bengbu Medical University, Bengbu 233000, China
  • Received:2025-04-14 Online:2025-12-20 Published:2025-12-22
  • Contact: Ying YU E-mail:13017518983@163.com;yuying2011@126.com

摘要:

目的 观察白藜芦醇(RES)对PM2.5诱导的脑缺血再灌注损伤小鼠血脑屏障的影响,并探讨线粒体分裂与融合在内皮屏障中的作用。 方法 将小鼠脑微血管内皮细胞分为4组:对照组(CON组)、模型组(OGD/R组)、实验组(OGD/R+PM2.5组)、RES组(OGD/R+PM2.5+RES组)。OGD/R+PM2.5组和RES组在OGD/R前先进行PM2.5(100 μg/mL)预处理24 h,RES组在复氧时更换含RES(40 mg/mL)的正常培养基培养18 h。对照组不给予任何处理。CCK-8检测细胞活性;跨内皮电阻(TEER)和FITC-Dextran评估细胞通透性;测定MDA含量和SOD活性;荧光探针检测细胞内及线粒体ROS水平;Mito-Tracker Red CMXRos检测线粒体形态;Western blotting检测细胞紧密连接蛋白(ZO-1、Occludin、Claudin-5)以及线粒体动力学相关蛋白(Drp1、Fis1、Mfn2、OPA1)表达水平。 结果 与对照组相比,OGD/R组及OGD/R+PM2.5组细胞TEER值降低、通透性增加,氧化应激水平升高,ROS荧光表达增强(P<0.05)。线粒体形态破碎不规则,紧密连接蛋白及线粒体融合蛋白表达降低,线粒体分裂蛋白表达升高(P<0.05)。RES干预后,可明显降低细胞膜通透性及ROS表达水平;改善线粒体形态,增加紧密连接蛋白与线粒体融合蛋白表达,降低分裂蛋白表达(P<0.05)。 结论 RES可通过调节线粒体动力学平衡,减轻PM2.5诱导的脑缺血再灌注血脑屏障的损伤,其机制可能与促进线粒体融合、抑制线粒体分裂有关。

关键词: PM2.5, 白藜芦醇, 脑缺血再灌注损伤, 线粒体分裂和融合, 血脑屏障

Abstract:

Objective To evaluate the effect of resveratrol (RES) on barrier function of mouse brain microvascular endothelial cell monolayers exposed to oxygen/glucose deprivation/reoxygenation (OGD/R) and PM2.5 and explore the role of mitochondrial fission and fusion in protecting endothelial barrier function. Methods Cultured mouse brain microvascular endothelial cells were exposed to OGD/R, treated with PM2.5 (100 μg/mL) before OGD/R, or pretreated with RES (40 mg/mL) prior to OGD/R+PM2.5 exposures. The changes in cell viability were examined with CCK-8 assay, and cell permeability was assessed by measuring transendothelial electrical resistance (TEER) and FITC-dextran permeation. Malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were measured, and intracellular and mitochondrial ROS levels were detected using fluorescent probes. Mitochondrial morphology in the treated cells was observed using Mito-Tracker Red CMXRos. Western blotting was performed to detect the changes in cellular expressions of the tight junction proteins (ZO-1, occludin, and claudin-5) and mitochondrial dynamics-associated proteins (Drp1, Fis1, Mfn2, and OPA1). Results Compared with the normal control cells, the cells exposed to OGD/R or both OGD/R and PM2.5 showed significantly decreased TEER levels, increased permeability, elevated oxidative stress, and increased ROS fluorescence intensities. Obvious mitochondrial fragmentation and morphological changes in the mitochondria were observed in the exposed cells, which also showed decreased expressions of tight junction proteins and mitochondrial fusion proteins with increased expressions of mitochondrial fission proteins. RES pretreatment of the endothelial cells before the exposures significantly reduced membrane permeability, lowered ROS levels, improved mitochondrial morphology, increased expressions of tight junction and fusion proteins, and decreased fission protein expressions. Conclusion RES can protect barrier function of mouse brain microvascular endothelial cell monolayers exposed to OGD/R and PM2.5 by modulating mitochondrial dynamics, potentially through promoting mitochondrial fusion and inhibiting mitochondrial fission.

Key words: PM2.5, resveratrol, cerebral ischemia-reperfusion injury, mitochondrial fission and fusion, Blood-brain barrier