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目的:通过3-溴丙酮酸(3-BrPA)处理秀丽隐杆线虫(Caenorhabditis elegans),观察3-BrPA对线虫的毒性和生存周期的影响.通过秀丽隐杆线虫RNA干扰(RNAi)和突变株,分析3-BrPA对线虫糖酵解途径己糖激酶家族和药物代谢的关键酶细胞色素P450(cytochrome P450,CYP)家族的影响.创新点:首次报道了3-BrPA对秀丽隐杆线虫有毒性作用,己糖激酶是3-BrPA对线虫作用的重要靶点,而CYP-35A家族是线虫代谢3-BrPA的主要酶类.方法:用不同浓度的3-BrPA处理秀丽隐杆线虫,每24 h监测一次存活率;用实时荧光定量聚合酶链反应(qPCR)检测代谢相关基因的表达;通过RNAi沉默己糖激酶家族基因hxk-1、hxk-2和hxk-3;计算3-BrPA处理hxk家族RNAi株和细胞色素P450 cyp-35家族突变株后的致死中浓度(LC50).结论:3-BrPA对线虫有明显的毒性效应(图1);与对照组比较,3-BrPA处理组的线虫平均寿命明显缩短(图2);3-BrPA处理线虫后hxk-1、hxk-2和hxk-3的信使RNA(mRNA)表达明显升高(P<0.05,图5);3-BrPA处理hxk RNAi株后的LC50均减小(P<0.05,表5);3-BrPA处理cyp-35突变株后的LC50也均减小(P<0.05,表6).综上所述,3-BrPA的毒性与其对秀丽隐杆线虫己糖激酶代谢的影响密切相关;CYP-35家族在线虫中对3-BrPA代谢中起着关键作用.“,”In this study, we aimed to evaluate the toxic effects, changes in life span, and expression of various metabolismrelated genes in Caenorhabditis elegans, using RNA interference (RNAi) and mutant strains, after 3-bromopyruvate (3-BrPA) treatment. C. elegans was treated with various concentrations of 3-BrPA on nematode growth medium (NGM) plates, and their survival was monitored every 24 h. The expression of genes related to metabolism was measured by the real-time fluorescent quantitative polymerase chain reaction (qPCR). Nematode survival in the presence of 3-BrPA was also studied after silencing three hexokinase (HK) genes. The average life span of C. elegans cultured on NGM with 3-BrPA was shortened to 5.7 d compared with 7.7 d in the control group. hxk-1, hxk-2, and hxk-3 were overexpressed after the treatment with 3-BrPA. After successfully interfering hxk-1, hxk-2, and hxk-3, the 50％ lethal concentration (LC50) of all mutant nematodes decreased with 3-BrPA treatment for 24 h compared with that of the control. All the cyp35 genes tested were overexpressed, except cyp-35B3. The induction of cyp-35A1 expression was most obvious. The LC50 values of the mutant strains cyp-35A1, cyp-35A2, cyp-35A4, cyp-35B3, and cyp-35C1 were lower than that of the control. Thus, the toxicity of 3-BrPA is closely related to its effect on hexokinase metabolism in nematodes, and the cyp-35 family plays a key role in the metabolism of 3-BrPA.