利用秀丽隐杆线虫这一与人类基因组高度同源的模式生物探究不同浓度的5-羟甲基糠醛(5-hydroxym-ethylfurfural,5-HMF)对机体的作用及其可能的机制.将实验分组设定为对照组和实验组(5-HMF质量浓度分别为0.010、0.100、1.000 mg/mL).通过显微镜观察线虫寿命、产卵情况、基础运动指标以及通过实时荧光定量PCR分析寿命信号通路:胰岛素/胰岛素样生长因子相关基因表达情况,并拟合出与寿命相关的量效关系.结果显示,高(1.000 mg/mL)、中(0.100 mg/mL)质量浓度组5-HMF可显著减少线虫产卵数量、咽泵频率、头部摆动和身体弯曲次数,并通过上调基因Daf-2的表达,下调基因Daf-16、Sod-3以及Sir-2.1的表达,显著缩短线虫寿命周期,而低(0.010 mg/mL)质量浓度组的5-HMF则完全相反.由量效关系曲线可知0.035 mg/mL是5-HMF影响线虫平均最长寿命的临界质量浓度值,0.056 mg/mL是5-HMF影响线虫最长寿命的临界质量浓度值.5-HMF作用于基因Daf-2、Daf-16、Sod-3以及Sir-2.1的临界质量浓度分别为0.067、0.067、0.075、0.052 mg/mL.结果表明,5-HMF对秀丽隐杆线虫的正、负向影响与浓度和作用时间密切相关,并存在一定量效关系.该文为进一步研究5-HMF对机体的影响提供了初步参考,并为5-HMF的毒理分析和安全应用提供理论依据.
Caenorhabditis elegans,a model organism highly homologous to the human genome,was used to explore the effects of dif-ferent concentrations of 5-hydroxymethylfurfural(5-HMF)on the organism and its possible mechanisms.The experimental groups includ-ed the control and experimental group(5-HMF concentrations were 0.010,0.100,and 1.000 mg/mL,respectively).The lifespan,egg production,and basal locomotor indexes of nematodes were observed by microscope;the lifespan signaling pathways,including insulin/in-sulin-like growth factor-related gene expression were analyzed by qRT-PCR,and the quantitative-effect relationships associated with lifes-pan were fitted.5-HMF in the high(1.000 mg/mL)and medium(0.100 mg/mL)concentration groups significantly decreased the num-ber of eggs laid,pharyngeal pump frequency,head swings,and body bends of nematodes,and significantly shortened the life cycle of nematodes by upregulating the expression of gene Daf-2 and downregulating the expression of genes Daf-16,Sod-3,and Sir-2.1,while the low(0.010 mg/mL)concentration group of 5-HMF was completely opposite.From the dose-effect relationship curve,it can be seen that the concentration of 0.035 mg/mL was the critical concentration value for 5-HMF to affect the average maximum lifespan of nematodes,and the concentration of 0.056 mg/mL was the critical concentration value for 5-HMF to affect the maximum lifespan of nematodes.The critical concentrations of 5-HMF acting on genes Daf-2,Daf-16,Sod-3,and Sir-2.1 were 0.067,0.067,0.075,and 0.052 mg/mL,re-spectively.The positive and negative effects of 5-HMF on C.elegans were closely related to the concentration and duration of action,and a dose-effect relationship existed.These results provide a preliminary reference for further research on the effects of 5-HMF on the organ-ism and provide a theoretical basis for the toxicological analysis and safe application of 5-HMF.