ALDH2通过线粒体融合与裂变减轻脂多糖诱导的脑微血管内皮细胞屏障的损伤  

ALDH2 attenuates LPS-induced increase of brain microvascular endothelial cell permeability by promoting fusion and inhibiting fission of the mitochondria

作　　者：王莎莎 吕恒 王丽娅 田美惠 高洁[1,2] 刘忠义 王佳慧 于影[1,2] WANG Shasha;LÜHeng;WANG Liya;TIAN Meihui;GAO Jie;LIU Zhongyi;WANG Jiahui;YU Ying(Department of Physiology,Bengbu Medical College,Bengbu 233000,China;Key Laboratory of Basic and Clinical Cardiovascular Diseases,Bengbu Medical College,Bengbu 233000,China;Department of Epidemiology and Health Statistics,Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院生理学教研室,安徽蚌埠233000 [2]蚌埠医学院心脑血管疾病基础与临床重点实验室,安徽蚌埠233000 [3]蚌埠医学院流行病与卫生统计学教研室,安徽蚌埠233000

出　　处：《南方医科大学学报》2022年第12期1882-1888,共7页Journal of Southern Medical University

基　　金：安徽省自然科学基金(2108085MH252);蚌埠医学院“512人才培育计划”(by51201307)。

摘　　要：目的观察乙醛脱氢酶2(ALDH2)对脂多糖(LPS)诱导的小鼠脑微血管内皮细胞屏障的影响,并探讨线粒体融合与裂变在内皮屏障中的作用。方法小鼠脑微血管内皮细胞分为3组:对照组:不给予任何处理;LPS损伤组(LPS):1μg/mL的LPS刺激24 h;LPS+ALDH2激动剂Alda-1组(LPS+Alda-1):在LPS刺激前给予20μmol/mL的Alda-1预处理1 h。CCK-8实验检测细胞活性;跨内皮细胞电阻和FITC-Dextran葡聚糖实验检测细胞通透性变化;试剂盒丙二醛(MDA)和超氧化物歧化酶(SOD)测定氧化应激水平;超氧化物阴离子荧光探针检测细胞活性氧(ROS)的表达;Western blot检测细胞ALDH2、紧密连接蛋白ZO-1、Occludin以及线粒体融合蛋白Mfn2、OPA1和分裂蛋白Drp1、Fis1表达。结果与对照组相比,LPS组跨内皮细胞电阻值降低、FITC-Dextran渗漏性增高,MDA含量升高、SOD活性下降,ROS荧光表达增强,ALDH2、紧密连接蛋白ZO-1、Occludin、线粒体融合蛋白Mfn2和OPA1表达降低,而线粒体分裂蛋白Drp1和Fis1表达升高(P<0.05)。应用Alda-1干预后,可显著抑制LPS所致的内皮细胞膜通透性升高,减低ROS荧光表达,ALDH2、ZO-1、Occludin、OPA1和Mfn2蛋白表达增高,而Drp1和Fis1蛋白表达降低(P<0.05)。结论ALDH2通过抑制线粒体ROS的产生,减轻LPS诱导脑微血管内皮细胞屏障的损伤,其机制可能与促进线粒体融合及抑制线粒体裂变有关。Objective To investigate the effect of aldehyde dehydrogenase 2(ALDH2)on lipopolysaccharide(LPS)-induced damage of mouse brain microvascular endothelial barrier and explore the role of mitochondrial fusion and fission in maintaining the integrity of endothelial barrier.Methods Mouse brain microvascular endothelial cells were treated with 1μg/mL LPS for 24 h with or without pretreatment with 20μmol/mL Alda-1(a ALDH2 agonist)for 1 h.The changes in cell viability were assessed using cell counting Kit-8(CCK8)assay,and the cell permeability was evaluated using transendothelial cell resistance(TEER)and FITC-Dextran assay.The level of oxidative stress in the cells was assessed by detecting the levels of malondialdehyde(MDA)and superoxide dismutase(SOD),and the content of reactive oxygen species(ROS)was detected using a superoxide anion fluorescent probe(DHE).Western blotting was performed to detect the expressions of ALDH2,tight junction proteins ZO-1 and occludin,and mitochondrial fusion-and division-related proteins Mfn2,OPA1,Drp1 and Fis1.Results Compared with the untreated cells,the cells treated with LPS showed significantly decreased TEER,increased FITC-dextran leakage,MDA content and ROS production,decreased SOD activity expressions of ALDH2,ZO-1,occludin,Mfn2 and OPA1,and increased expressions of Drp1 and Fis1(P<0.05).Pretreatment with Alda-1 prior to LPS exposure strongly suppressed the increase of endothelial cell membrane permeability,reduced ROS production,increased the expressions of ALDH2,ZO-1,occludin,OPA1 and Mfn2,and lowered the expressions of Drp1 and Fis1(P<0.05).Conclusion ALDH2 can alleviate LPS-induced damage of brain microvascular endothelial cell barrier by inhibiting the mitochondrial ROS production and promoting mitochondrial fusion and inhibiting mitochondrial fission.

关 键 词：乙醛脱氢酶2 LPS 线粒体融合和分裂 内皮屏障 

分 类 号：R459.7[医药卫生—急诊医学] R747.9[医药卫生—治疗学]