5-羟甲基糠醛通过活性氧和Wnt信号通路影响斑马鱼幼鱼心血管发育研究  

Effects of 5-hydroxymethylfurfural on cardiovascular development in zebrafish larvae via reactive oxygen species and Wnt signaling pathways

作　　者：冯诗缘 吴雯 史静玲 姜宇 Feng Shiyuan;Wu Wen;Shi Jingling;Jiang Yu(Department of General Practice,the Affiliated Wuxi Second People’s Hospital of Nanjing Medical University,Wuxi 214002,China)

机构地区：[1]南京医科大学附属无锡第二医院全科医学,无锡214002

出　　处：《中华实验外科杂志》2023年第8期1526-1530,共5页Chinese Journal of Experimental Surgery

基　　金：中国博士后科学基金会(2019T120461)。

摘　　要：目的探索5-羟甲基糠醛(5-HMF)影响心血管发育的机制。方法以10、25、50 μg/ml的5-HMF处理受精8 h后的AB系斑马鱼幼鱼, 用E3溶液处理同胞幼鱼作为对照组, 进行形态组织学观察, 吖啶橙染色检测细胞凋亡, 实时定量聚合酶链反应(RT-qPCR)检测相关基因表达。用抗氧化剂N-乙酰-l-半胱氨酸(NAC)和Wnt信号通路激活剂(BML)处理幼鱼进行挽救实验, 比较各组幼鱼心血管发育情况。数据比较采用单因素方差分析。结果高浓度5-HMF处理组幼鱼心率、心包水肿程度、静脉窦-动脉球距、凋亡染色面积[(76.23±2.59)次/20 s、(65.83±3.12)%、(18.38±2.33) μm、(8 639.28±798.06)]均高于对照组[(14.39±3.41)次/20 s、(0.84±0.12)%、(14.39±0.76) μm、(1 420.84±359.34)], 差异有统计学意义(t=9.312、8.590、11.784、9.786, P<0.05)。高浓度5-HMF处理组幼鱼心脏发育相关基因心脏特异性同源盒转录因子(nkx2.5)、T-框蛋白5(Tbx5)、锌指转录因子(gata4)和血管发育相关基因血管内皮生长因子A旁系同源基因(VEGF-A)的表达水平(0.48±0.06、0.34±0.04、0.64±0.08、0.78±0.13)均低于对照组(1.00±0.12、1.00±0.17、1.00±0.13、1.00±0.18), 差异有统计学意义(t=5.624、4.439、5.804、6.872, P<0.05)。而凋亡相关基因B细胞淋巴瘤/白血病-2相关X蛋白(bax)和p53的表达水平(2.56±0.36、3.65±0.48)均高于对照组(1.00±0.18、1.00±0.13), 差异有统计学意义(t=7.283、6.392, P<0.05)。NAC和BML可以挽救5-HMF引起的表型异常。结论高浓度5-HMF提高了斑马鱼幼鱼活性氧水平, 抑制Wnt信号通路的转导, 最终导致斑马鱼幼鱼心血管发育异常。Objective To investigate the cardiovascular effects of 5-hydroxymethylfurfural(5-HMF).Methods At 8 h after fertilization,embryos were treated with 10,25 and 50μg/ml 5-HMF,and controls were cultured with E3,followed by histological observation,fluorescence observation,cell apoptosis staining,and gene quantitative analysis.The larvae were treated with N-acetyl-L-cysteine(NAC)and Wnt signaling pathway activator(BML)to carry on rescue experiment.The statistical method was one-way analysis of variance.Results The heart rate,pericardial edema ratio,venous sinus-arterial bulb distance and pericardial apoptotic staining area in high 5-HMF treatment groups[(76.23±2.59)times/20 s,(65.83±3.12)%,(18.38±2.33)μm,(8639.28±798.06)]significantly increased as compared with those in the control group[(14.39±3.41)times/20 s,(0.84±0.12)%,(14.39±0.76)μm,(1420.84±359.34,t=9.312,8.590,11.784,9.786,P<0.05].The expression levels of the cardiac development-related genes nk2 homeobox 5(nkx2.5),T-box protein 5(Tbx5),GATA binding factor-4(gata4)and the vascular development-related gene vascular endothelial growth factor A(VEGF-A)(0.48±0.06,0.34±0.04,0.64±0.08,0.78±0.13)in the high 5-HMF treatment groups were lower than those in the control group(1.00±0.12,1.00±0.17,1.00±0.13,1.00±0.18,t=5.624,4.439,5.804,6.872,P<0.05).The expression levels of apoptosis-related genes B cell lymphoma/leukemia-2 associated X protein(bax)and p53(2.56±0.36,3.65±0.48)in the high 5-HMF treatment groups were higher than those in the control group(1.00±0.18,1.00±0.13,t=7.283,6.392,P<0.05).The abnormal phenotype caused by 5-HMF can be rescued by NAC and BML.Conclusion It is inferred that high 5-HMF concentrations increased the level of reactive oxygen species,inhibited the transduction of the Wnt signaling pathway,and resulted in abnormal cardiovascular development in zebrafish larvae.

关 键 词：5-羟甲基糠醛 心脏毒性 血管 活性氧自由基 斑马鱼 

分 类 号：R54[医药卫生—心血管疾病]