3-溴丙酮酸对秀丽隐杆线虫的毒性和代谢的影响（英文）
发布时间：2021-04-29 13:43
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-7、hxk-2和hxk-3;计算3-BrPA处理hxk家族RNAi株和细胞色素P450cym-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RN...
【文章来源】：Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2020,21(01)SCICSCD
【文章页数】：10 页
【文章目录】：
1 Introduction
2 Materials and methods
2.1 Cultivation of C.elegans
2.2 Exposure of C.elegans to 3-BrPA
2.3 Construction of HK gene molecular phyloge-netic tree
2.4 Total RNA extraction and reverse transcrip-tion of C.elegans
2.5 Determination of hxk-1,hxk-2,and hxk-3m RNA expression
2.6 Silencing of the target genes hxk-1,hxk-2,and hxk-3
2.6.1 Cloning the target genes
2.6.2 Expression of the target genes
2.6.3 Target gene silencing
2.6.4 Design of CYP35 family primers and deter-mination of expression of cyp35 genes
2.7 Action of 3-BrPA on RNAi or mutant nema-tode strains
2.8 Data analysis
3 Results
3.1 Effect of 3-BrPA on the life span of nematodes
3.2 Evolutionary genetic comparison of the HK genes
3.3 mRNA expression of the HK genes in C.ele-gans treated with 3-BrPA
3.4 Effect of 3-BrPA on the HK genes in RNAi nematodes
3.5 Effect of 3-BrPA on the expression of the cyp-35 gene in mutant strains
3.6 LC50 of 3-BrPA on the cyp-35 gene mutant strains
4 Discussion
5 Conclusions
Contributors
Compliance with ethics guidelines
本文编号：3167635
【文章来源】：Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2020,21(01)SCICSCD
【文章页数】：10 页
【文章目录】：
1 Introduction
2 Materials and methods
2.1 Cultivation of C.elegans
2.2 Exposure of C.elegans to 3-BrPA
2.3 Construction of HK gene molecular phyloge-netic tree
2.4 Total RNA extraction and reverse transcrip-tion of C.elegans
2.5 Determination of hxk-1,hxk-2,and hxk-3m RNA expression
2.6 Silencing of the target genes hxk-1,hxk-2,and hxk-3
2.6.1 Cloning the target genes
2.6.2 Expression of the target genes
2.6.3 Target gene silencing
2.6.4 Design of CYP35 family primers and deter-mination of expression of cyp35 genes
2.7 Action of 3-BrPA on RNAi or mutant nema-tode strains
2.8 Data analysis
3 Results
3.1 Effect of 3-BrPA on the life span of nematodes
3.2 Evolutionary genetic comparison of the HK genes
3.3 mRNA expression of the HK genes in C.ele-gans treated with 3-BrPA
3.4 Effect of 3-BrPA on the HK genes in RNAi nematodes
3.5 Effect of 3-BrPA on the expression of the cyp-35 gene in mutant strains
3.6 LC50 of 3-BrPA on the cyp-35 gene mutant strains
4 Discussion
5 Conclusions
Contributors
Compliance with ethics guidelines
本文编号：3167635
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