临床儿科杂志 ›› 2015, Vol. 33 ›› Issue (3): 269-.doi: 10.3969j.issn.1000-3606.2015.03.018

• 实验研究 • 上一篇    下一篇

牛磺酸对缺氧再灌注斑马鱼幼鱼脑部内质网应激相关因子表达的影响

罗济璇,程燕,赵丹,陈衍晨,王斌   

  1. 南方医科大学珠江医院儿科(广东广州 510280)
  • 收稿日期:2015-03-15 出版日期:2015-03-15 发布日期:2015-03-15
  • 通讯作者: 王斌 E-mail:wangbin6556@126.com

Effect of taurine on the expression of endoplasmic reticulum stress-related factors in the brain of zebrafish larvae after hypoxia reperfusion 

LUO Jixuan, CHENG Yan, ZHAO Dan, CHEN Yanchen, WANG Bin   

  1. Department of Pediatrics, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong, China
  • Received:2015-03-15 Online:2015-03-15 Published:2015-03-15

摘要: 目的 观察牛磺酸对缺氧再灌注斑马鱼幼鱼脑组织中内质网应激病理状态标志物葡萄糖调节蛋白78(GRP78)、内质网应激特异性下游蛋白C/EBP同源蛋白(CHOP)及半胱氨酸天冬氨酸蛋白酶-12(caspase-12)表达及神经元凋亡的影响,探讨其脑保护机制。方法 将受精后5 d的斑马鱼幼鱼随机分为空白对照组、缺氧再灌注模型组以及牛磺酸组,其中牛磺酸组按不同浓度进一步分为1 mmol/L、5 mmol/L、10 mmol/L 3个亚组,每组均100条。观察并记录缺氧后再灌注1 h的斑马鱼幼鱼的行为、苏醒时间、中位生存时间、尼氏染色及原位末端标记法检测神经元凋亡情况,Western blot法测各组幼鱼脑部GRP78、CHOP及caspase-12表达水平。结果 与模型组相比,各牛磺酸处理组的苏醒时间缩短、中位生存时间延长、尼氏染色阳性神经元计数增多以及凋亡神经元计数减少,差异均有统计学意义(P<0.05);GRP78、CHOP和caspase-12在模型组及各牛磺酸处理组均有表达,但三者在各牛磺酸处理组的表达均较模型组低,差异有统计学意义(P<0.05)。结论 缺氧再灌注诱发内质网应激,牛磺酸可能通过下调GRP78、CHOP和caspase-12减少神经元凋亡从而减轻斑马鱼幼鱼缺氧再灌注脑损伤。

Abstract: Objectives To investigate the expression of glucose regulated protein 78 (GRP78), CCAAT/enhancer binding
protein homologous protein (CHOP) and cysteine asparate protease-12 (caspase-12) and neuronal apoptosis in the brain of
zebrafish larvae after hypoxia reperfusion, and the neuroprotective effect of taurine. Methods  Five day old post-fertilization zebrafish
 larvae were randomly assigned into 3 groups, control group, hypoxia reperfusion group (model group) and taurine group, and the taurine group was further divided into 3 subgroups according to different concentrations (1 mmol/L, 5 mmol/L, 10 mmol/L) with 100 zebrafish larvae each. The behavior, recovery time and median survival time of those zebrafish larvae after hypoxia with 1h reperfusion were observed and recorded. The pathological changes and apoptosis of neurons were detected by Nissl staining and terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling. The expression of GRP78, CHOP and caspase-12 in the brain of zebrafish larvae were detected by Western blot. Results  Compared with the model group, the recovery time was shortened, the median survival time was extended, the Nissl stained neurons was increased and the apoptotic neurons were
decreased in the taurine groups. GRP78, CHOP and caspase-12 were expressed in model group and taurine group. The expression of GRP78, CHOP and caspase-12 was much lower in taurine group than in model group. Conclusions  Hypoxia reperfusion may induce endoplasmic reticulum stress and taurine may be neuroprotective against hypoxia reperfusion by down-regulating GRP78, CHOP and caspase-12.