丙泊酚对不同发育时期斑马鱼鞘磷脂蛋白表达的影响

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

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30μg/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。

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 30μg/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.