南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (09): 1115-.doi: 10.12122/j.issn.1673-4254.2018.09.15

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胚胎期斑马鱼丙泊酚暴露可下调髓鞘碱性蛋白的表达

刘川，林春水，郭培培，张昕，朱晓勤   

  • 出版日期:2018-09-20 发布日期:2018-09-20

Exposure to propofol down-regulates myelin basic protein expression in zebrafish embryos: its neurotoxicity on oligodendrocytes and the molecular mechanisms

  • Online:2018-09-20 Published:2018-09-20

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摘要： 目的探讨胚胎期斑马鱼丙泊酚暴露下调髓鞘碱性蛋白（MBP）表达及相关机制。方法受精后6~48 h（6~48 hpf）的斑马 鱼胚胎，随机分为对照组、二甲基亚砜（DMSO）组、20 μg/mL丙泊酚组和30 μg/mL丙泊酚组，采用浸泡方式给药。于48 hpf、 72 hpf 收集幼鱼。通过荧光定量RT-qPCR法检测MBP mRNA和少突胶质细胞转录因子1（Olig1）、Olig2、性别决定区盒基因 （Sox10）mRNA表达（n=80条/组）；Western blot法检测MBP蛋白表达（n=80条/组）；TUNEL、免疫荧光法检测少突胶质细胞凋亡 （n=6条/组）。结果与对照组相比，20 μg/mL、30 μg/mL丙泊酚暴露两组MBP mRNA、MBP蛋白于72 hpf均明显下调（P<0.05）， 另外30 μg/mL 组较20 μg /mL 组MBP蛋白表达显著减少（P<0.05）；20 μg /mL、30 μg /mL 两组转录因子Olig1、Olig2、Sox10 mRNA表达于48 hpf、72 hpf明显下调（P<0.05），少突胶质细胞凋亡于72 hpf显著增多（P<0.05）。结论胚胎期斑马鱼丙泊酚暴 露可能通过诱导少突胶质细胞凋亡增多及抑制转录因子Olig1、Olig2、Sox10，从而下调MBP表达。

Abstract: Objective To investigate the mechanism underlying propofol- induced down-regulation of myelin basic protein (MBP) in zebrafish embryos. Methods Zebrafish embryos (6-48 h post-fertilization [hpf]) were randomized into 4 equal groups for exposure to dimethyl sulfoxide (DMSO), 20 μg/mL propofol, 30 μg/mL propofol, or no particular treatment (control group). The larvae were collected at 48 or 72 hpf for detecting the mRNA levels of MBP, Olig1, Olig2, and Sox10 using qRT-PCR (n=80). The protein expression of MBP was quantitatively detected using Western blotting (n=80), and the apoptosis of the oligodendrocytes was investigated using TUNEL staining (n=6). Results Exposure to 20 and 30 μg/mL propofol caused significant reductions in the mRNA expressions of Olig1, Olig2, and Sox10 at 48 and 72 hpf (P<0.05) and also in MBP mRNA and protein levels at 72 hpf (P<0.05). Exposure to 30 μg/mL propofol induced more obvious reduction in MBP protein expression than 20 μg/mL propofol at 72 hpf (P<0.05), and the exposures resulted in a significant increase of oligodendrocyte apoptosis at 72 hpf (P<0.05). Conclusion Propofol exposure reduces MBP expression at both the mRNA and protein levels in zebrafish embryos by down-regulating the expressions of Olig1, Olig2 and Sox10 mRNA levels and increasing apoptosis of the oligodendrocytes.