南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1810-1814.doi: 10.12122/j.issn.1673-4254.2023.10.21

• • 上一篇    下一篇

丙泊酚对不同发育时期斑马鱼鞘磷脂蛋白表达的影响

郭志华,王志鹏,曾琳玲,纪雪霞   

  1. 南方医科大学附属广东省人民医院(广东省医学科学院)麻醉科,广东 广州 510080
  • 出版日期:2023-10-20 发布日期:2023-11-02

Effects of propofol on myelin basic protein expression in zebrafish at different developmental stages

GUO Zhihua, WANG Zhipeng, ZENG Linling, JI Xuexia   

  1. Department of Anesthesiology, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou 510080, China
  • Online:2023-10-20 Published:2023-11-02

摘要: 目的 探讨丙泊酚对斑马鱼鞘磷脂蛋白(MBP)表达的影响及相关机制。方法 将受精后6~48 h胚胎(共180个)采用随机数字表法分别浸泡于新鲜养鱼水(对照组)、含二甲基亚砜养鱼水(二甲基亚砜组)和含30 μg/mL丙泊酚养鱼水(丙泊酚组)中,每组60个胚胎。于受精后3、4、5、6、7、10 d收集幼鱼,通过RT-qPCR法和Western blot法检测不同发育时期mbp mRNA及蛋白表达。采用TUNEL、免疫荧光法检测3 d少突胶质细胞的凋亡情况。采用RT-qPCR和Western blot法检测凋亡相关因子caspase-8、caspase-9和caspase-3的表达。结果 与对照组相比,丙泊酚组出生后3~7 d幼鱼mbp mRNA和蛋白表达均降低(P<0.05);丙泊酚组少突胶质细胞凋亡增多,并伴随caspase-8、caspase-9和caspase-3 mRNA和蛋白的表达上调。结论 丙泊酚抑制MBP表达具有短期持续性,其机制可能与少突胶质细胞凋亡相关。

关键词: 斑马鱼;少突胶质细胞;鞘磷脂蛋白;细胞凋亡

Abstract: Objective To observe the effect of propofol on the expression of myelin basic protein (MBP) in developing zebrafish and explore the possible mechanisms. Methods A total of 180 zebrafish embryos at 6-48 h post- fertilization were randomly allocated into 3 equal groups and raised in fresh water (control group), water containing dimethyl sulfoxide (DMSO group) and water containing 30 μg/mL propofol (propofol group). On 3, 4, 5, 6, 7, 10 d post-fertilization, the juvenile fish were collected for detection of mRNA and protein expressions of MBP using RT- qPCR and Western blotting. TUNEL assay and immunofluorescence assay were used to detect apoptosis of the oligodendrocytes of the fish at 3 d post-fertilization; RT-qPCR and Western blotting were performed to detect the expressions of apoptosis-related factors caspase-8, caspase-9 and caspase-3. Results Compared with the control group, the fish with propofol exposure showed significantly decreased mRNA and protein expression of MBP at 3-7 d post-fertilization (P<0.05) with increased apoptosis of the oligodendrocytes and upregulated expressions of caspase-8, caspase-9 and caspase-3 at both the mRNA and protein levels. Conclusion Propofol persistently inhibits MBP expression in developing zebrafish within a short term possibly by mediating apoptosis of the oligodendrocytes.

Key words: zebrafish; oligodendrocyte; myelin basic protein; cell apoptosis