Full Text:   <1817>

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CLC number: R965.3

On-line Access: 2020-01-03

Received: 2019-07-12

Revision Accepted: 2019-09-04

Crosschecked: 2019-12-17

Cited: 0

Clicked: 2657

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Gen Chen

https://orcid.org/0000-0002-5780-0901

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Journal of Zhejiang University SCIENCE B 2020 Vol.21 No.1 P.77-86

http://doi.org/10.1631/jzus.B1900370


Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans


Author(s):  Qiao-ling Gu, Yan Zhang, Xi-mei Fu, Zhao-lian Lu, Yao Yu, Gen Chen, Rong Ma, Wei Kou, Yong-mei Lan

Affiliation(s):  The Institute of Minority Physique and Health, Medical College of Northwest University for Nationalities, Lanzhou 730030, China; more

Corresponding email(s):   chengen1999@163.com

Key Words:  3-Bromopyruvate, Caenorhabditis elegans, Hexokinase, Cytochrome P450


Qiao-ling Gu, Yan Zhang, Xi-mei Fu, Zhao-lian Lu, Yao Yu, Gen Chen, Rong Ma, Wei Kou, Yong-mei Lan. Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans[J]. Journal of Zhejiang University Science B, 2020, 21(1): 77-86.

@article{title="Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans",
author="Qiao-ling Gu, Yan Zhang, Xi-mei Fu, Zhao-lian Lu, Yao Yu, Gen Chen, Rong Ma, Wei Kou, Yong-mei Lan",
journal="Journal of Zhejiang University Science B",
volume="21",
number="1",
pages="77-86",
year="2020",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1900370"
}

%0 Journal Article
%T Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans
%A Qiao-ling Gu
%A Yan Zhang
%A Xi-mei Fu
%A Zhao-lian Lu
%A Yao Yu
%A Gen Chen
%A Rong Ma
%A Wei Kou
%A Yong-mei Lan
%J Journal of Zhejiang University SCIENCE B
%V 21
%N 1
%P 77-86
%@ 1673-1581
%D 2020
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1900370

TY - JOUR
T1 - Toxicity and metabolism of 3-bromopyruvate in Caenorhabditis elegans
A1 - Qiao-ling Gu
A1 - Yan Zhang
A1 - Xi-mei Fu
A1 - Zhao-lian Lu
A1 - Yao Yu
A1 - Gen Chen
A1 - Rong Ma
A1 - Wei Kou
A1 - Yong-mei Lan
J0 - Journal of Zhejiang University Science B
VL - 21
IS - 1
SP - 77
EP - 86
%@ 1673-1581
Y1 - 2020
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1900370


Abstract: 
In this study, we aimed to evaluate the toxic effects, changes in life span, and expression of various metabolism-related 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.

3-溴丙酮酸对秀丽隐杆线虫的毒性和代谢的影响

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-1hxk-2hxk-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-1hxk-2hxk-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.

关键词:3-溴丙酮酸;秀丽隐杆线虫;己糖激酶;细胞色素P450

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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