线粒体自噬对新生大鼠缺氧缺血脑损伤的影响

发布时间：2018-06-24 23:08

本文选题：线粒体自噬 + 缺氧缺血脑损伤　；参考：《中国当代儿科杂志》2017年02期

【摘要】：目的了解缺氧缺血脑损伤的动物模型中线粒体自噬情况,探讨其在缺氧缺血脑损伤中的作用。方法将120只新生7日龄Sprague-Dawley大鼠分为假手术组、缺氧缺血脑损伤(HIBD)组、自噬抑制剂干预组(3MA组)。HIBD组行右侧颈总动脉结扎后置于低氧仓(8%氧气和92%氮气)2.5 h,3MA组腹腔注射2μL 3MA 30 min后再进行结扎和低氧处理,假手术组不予结扎和低氧处理。在造模后提取各组单细胞悬液,用Mitotracker标记线粒体、Lysotracker标记自噬小体、LC3标记自噬进行免疫荧光共定位观察线粒体自噬情况;荧光探针JC-1染色后,采用流式细胞仪检测线粒体膜电位。TTC染色法检测脑梗死灶大小。提取皮质神经元中的细胞浆蛋白,Western blot法检测线粒体自噬相关蛋白的表达情况。结果与假手术组相比,HIBD组线粒体膜电位明显降低(P0.05),线粒体自噬增加(P0.05),线粒体分裂相关蛋白Drp1、Fis1的表达升高,线粒体外膜蛋白受体Tom20与内膜蛋白受体Tim23的表达降低(P0.05);3MA组膜电位比HIBD组降低更为明显(P0.05),但线粒体自噬程度明显减少(P0.05)。3MA组大鼠脑组织梗死程度较HIBD组明显增加(P0.05)。结论 HIBD能够增加线粒体自噬的发生,抑制线粒体自噬会加重新生大鼠HIBD的损伤程度。
[Abstract]:Objective to investigate the role of mitochondrial autophagy in hypoxic-ischemic brain injury model. Methods 120 7 day old Sprague-Dawley rats were divided into sham-operation group and hypoxic-ischemic brain injury (HIBD) group. The right common carotid artery was ligated and placed in hypoxia chamber (8% oxygen and 92% nitrogen) for 2.5 h in the autophagy inhibitor intervention group (3MA group) .HIBD group was treated with ligation and hypoxia after intraperitoneal injection of 2 渭 L 3MA for 30 min. After modeling, single cell suspension was extracted from each group, and mitochondrial Lysotracker labeled autophagy was used to observe the mitochondrial autophagy by immunofluorescence, and the fluorescence probe JC-1 staining was used to observe the mitochondrial autophagy. Mitochondrial membrane potential. TTC staining was used to detect the size of cerebral infarction by flow cytometry. The expression of mitochondrial autophagy associated protein was detected by Western blot method. Results compared with the sham-operated group, the mitochondrial membrane potential decreased significantly (P0.05), the mitochondrial autophagy increased (P0.05), and the expression of mitochondrial mitogen-associated protein Drp1 Fis1 increased in the HIBD group. The expression of mitochondrial outer membrane protein receptor Tom20 and intimal protein receptor Tim23 were significantly decreased (P0.05). The membrane potential of 3MA group was significantly lower than that of HIBD group (P0.05), but the degree of mitochondrial autophagy was significantly decreased (P0.05) .3MA group was significantly higher than HIBD group (P0.05). Conclusion HIBD can increase the occurrence of mitochondrial autophagy and inhibit mitochondrial autophagy can aggravate the damage of HIBD in neonatal rats.
【作者单位】：四川大学华西第二医院儿科/出生缺陷与相关妇儿疾病教育部重点实验室/发育与妇儿疾病四川省重点实验室;
【基金】：国家自然科学基金(81330016;81630038;81300526) 国家重点基础发展计划(973计划)(2013CB967404) 四川省科技计划项目(2014SZ0149) 临床重点专科计划项目(1311200003303)
【分类号】：R742

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