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线粒体自噬相关蛋白在新生鼠缺氧缺血皮质神经细胞中的表达

发表时间:2017-03-15  浏览量:1384  下载量:307
全部作者: 李明熹,张莉,屈艺,母得志
作者单位: 四川大学华西第二医院儿科;四川大学出生缺陷与相关妇儿疾病教育部重点实验室
摘 要: 目的:探讨线粒体自噬相关蛋白在缺氧缺血大鼠皮质神经细胞中的表达,了解在缺氧缺血脑损伤(hypoxic-ischemic brain damage,HIBD)动物模型中线粒体自噬情况。方法:60只雄性7日龄新生SD大鼠随机分为假手术组(sham组)、缺氧缺血脑损伤组(HIBD组)、自噬抑制剂干预组(3-MA组)。在缺氧缺血后2,12,24,48,72 h时,提取皮质神经元中的细胞浆蛋白和线粒体蛋白,采用Western blotting检测线粒体自噬相关蛋白的表达。结果:在细胞浆中,HIBD组LC3II/I、PINK1和Parkin的表达明显增高,与sham组相比差异显著(P<0.01),在手术12 h后表达开始升高,48 h后开始下降,72 h后逐渐恢复正常;线粒体外膜分裂相关蛋白Drp1、Fis1的表达与PINK1、Parkin一致,在12~24 h表达升高,HIBD组各时间点与sham组相比存在明显差异(P<0.01),同时,线粒体外膜蛋白Tom20与线粒体内膜蛋白Tim23的表达在手术后逐渐降低,12~24 h表达最低,与sham组相比,差异有统计学意义(P<0.01);自噬抑制剂干预后,与sham组及HIBD组各时间点相比,PINK1、Parkin、Drp1、Fis1的表达降低,Tom20和Tim23的表达升高,差异均有统计学意义(P<0.01)。结论:线粒体自噬相关蛋白在HIBD后的皮质神经元线粒体中的表达具有一定的时间规律性,抑制线粒体自噬可能与HIBD的修复存在一定关系。
关 键 词: 儿科学;线粒体自噬;缺氧缺血;脑损伤
Title: Expression of mitophagy related protein in cortical neurons of hypoxic-ischemic newborn rat
Author: LI Mingxi, ZHANG Li, QU Yi, MU Dezhi
Organization: Department of Pediatrics, West China Second University Hospital, Sichuan University; Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of Education, Sichuan University
Abstract: Objective: To investigate the expression of mitophagy related protein in cortical neurons of hypoxic-ischemic newborn rats and to study the situation of mitophagy in the model of hypoxic-ischemic brain damage (HIBD). Methods: Sixty seven-day-old neonatal male SD rats were randomly divided into three groups: sham-operated group (sham), HIBD model group (HIBD), 3-MA injection group (3-MA). After 2, 12, 24, 48, 72 h of hypoxic-ischemic treatment, the protein in cortical neurons was extracted. Western blotting was used to measure the expression of mitophagy related protein. Results: In cytoplasm, compared with the sham group, the expression of LC3II/I, PINK1 and Parkin in HIBD group were significantly increased (P<0.01) after 12 h of hypoxic-ischemic treatment, decreased after 48 h and back to base-line at 72 h. The same tendency can be observed in the expression of mitochondrial outer membrane fission related protein (P<0.01). Simultaneously, the expression of mitochondrial membrane protein, Tom20 and Tim23, were reduced after surgery, which reached the lowest expression after 12-24 h (P<0.01), compared with sham group. After injecting of 3-MA, an inhibitor of mitophagy, the expression of PINK1/Parkin and Drp1/Fis1 were reduced, oppositely, the expression of Tom20/Tim23 were increased (P<0.01), compared with sham group and HIBD group at each time point. Conclusion: The expression of mitophagy related protein in cortical neurons of HIBD is on a regular or periodic basis, which may have some relationship with the protection role from HIBD.
Key words: pediatrics; mitophagy; hypoxic-ischemic; brain damage
发表期数: 2017年3月第5期
引用格式: 李明熹,张莉,屈艺,等. 线粒体自噬相关蛋白在新生鼠缺氧缺血皮质神经细胞中的表达[J]. 中国科技论文在线精品论文,2017,10(5):509-517.
 
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