肝缺血再灌注大鼠血清外泌体引起海马和大脑皮质损伤
张丽梅¹, 贾莉莉², 喻文立^2,*
1天津医科大学一中心临床学院，天津 300070；²天津市第一中心医院麻醉科，天津 300192
摘要
本文旨在探究大鼠肝缺血再灌注(ischemia/reperfusion, I/R)后血清中外泌体的变化及其在海马和大脑皮质损伤中的作用。采用随机数字表法将清洁级雄性Sprague-Dawley (SD)大鼠分为4组：假手术组(S组)、肝缺血再灌注组(I/R组)、S组大鼠血清外泌体处理组(ES组)和I/R组大鼠血清外泌体处理组(EI组)。ES组和EI组为正常大鼠经尾静脉分别注射S组和I/R组大鼠血清中的外泌体各100 μL。另取3只正常大鼠经尾静脉注射PKH26红色荧光标记的外泌体，利用免疫荧光显微镜观察大鼠脑组织中荧光表达情况。应用透射电子显微镜观察外泌体的形态和大小；免疫蛋白印迹法检测外泌体标记蛋白CD63和CD9的表达情况；通过血清学和组织学指标的检测，观察肝及脑组织损伤情况及海马和大脑皮质组织细胞凋亡和氧化应激水平。结果显示：外泌体是直径为30~100 nm的圆形或椭圆形膜性囊泡。与S组比较，I/R组大鼠血清中外泌体含量增加(P < 0.05)。外泌体具有血脑屏障通透性，可自外周经血液循环进入脑组织。I/R组血清肝功能指标较S组明显升高(P < 0.05)。与S组比较，ES组脑组织损伤程度及海马和大脑皮质组织细胞凋亡和氧化应激水平无显著变化；与S组、ES组比较，I/R组、EI组血清脑损伤标志物水平、海马和大脑皮质组织中凋亡指数、氧化应激水平升高，但EI组以上指标均低于I/R组(P < 0.05)。因此，大鼠肝I/R可导致海马和大脑皮质组织损伤，而且其介导的血清外泌体含量增加在其中发挥了重要作用。
关键词： 外泌体; 缺血再灌注; 肝; 海马; 大脑皮质
分类号：R333.4; R338

Serum exosomes of rats with hepatic ischemia-reperfusion injury induce the damage of hippocampus and cerebral cortex
ZHANG Li-Mei¹, JIA Li-Li², YU Wen-Li^2,*
1First Central Clinical College, Tianjin Medical University, Tianjin 300070, China；²Department of Anesthesiology, Tianjin First Central Hospital, Tianjin 300192, China
Abstract
The paper was aimed to explore the role of serum exosomes induced by hepatic ischemia/reperfusion (I/R) injury in the damage of hippocampus and cerebral cortex of rats. The male Sprague-Dawley (SD) rats were randomly divided into 4 groups: sham operation group (S), hepatic I/R injury group (I/R), serum exosomes from S group treatment group (ES) and serum exosomes from I/R group treatment group (EI). In ES group and EI group, 100 μL serum exosomes from S group and I/R group were injected into the normal rats through tail vein respectively. Another three normal rats were injected intravenously with serum exosomes labeled with PKH26 red fluorescence, and then the expression of fluorescence in the brain tissues was observed by immunofluorescence microscope. The morphology and size of exosomes were observed by transmission electron microscope, the expression of exosomes markers CD63 and CD9 was detected by Western blot, and the damage of liver and brain, levels of apoptosis and oxidative stress response in hippocampus and cerebral cortex were observed by serological and histological indexes. The results showed that the exosomes were a group of round or ovoid membranous vesicles, sized in 30–100 nm. Compared with that in S group, the content of serum exosomes in I/R group was increased (P < 0.05). Moreover, serum exosomes could go through the blood-brain barrier and enter the brain tissue freely through blood circulation. The index of liver function in I/R group was significantly higher than that in S group (P < 0.05). There was no significance in the degree of brain damage, apoptosis and oxidative stress in hippocampus and cerebral cortex between S group and ES group. Compared with those in S group and ES group, the serum levels of brain injury markers, apoptosis index (AI) and oxidative stress in hippocampus and cerebral cortex increased in I/R group and EI group (P < 0.05). Whereas, compared with those in I/R group, the above indicators in EI group decreased (P < 0.05). Therefore, hepatic I/R injury can lead to the damage of hippocampus and cerebral cortex, and the increased serum exosomes induced by hepatic I/R plays an important role.
Key words: exosomes; ischemia/reperfusion injury; liver; hippocampus; cerebral cortex

收稿日期：2019-09-19　　录用日期：2020-04-17
通讯作者：喻文立　　E-mail: yzxyuwenli@163.com
DOI: 10.13294/j.aps.2020.0044
引用本文：
张丽梅, 贾莉莉, 喻文立. 肝缺血再灌注大鼠血清外泌体引起海马和大脑皮质损伤[J]. 生理学报 2020; 72 (4): 449-454.
ZHANG Li-Mei, JIA Li-Li, YU Wen-Li. Serum exosomes of rats with hepatic ischemia-reperfusion injury induce the damage of hippocampus and cerebral cortex. Acta Physiol Sin 2020; 72 (4): 449-454 (in Chinese with English abstract).



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