抗生素的胁迫与抗生素抗性基因产生与传播关系的研究
The Relationship of Antibiotic Stress with Emergence and Dissemination of Antibiotic Resistance Genes
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摘要: 抗生素的环境残留和抗生素抗性基因（antibiotic resistance genes，ARGs）污染日益增加，对全球公共卫生构成重大威胁。目前，关于环境中抗生素与ARGs产生与传播的关系的研究较多，但结果却不尽相同。为了确定抗生素的胁迫与ARGs产生与传播的关系，用磺胺类抗生素（SAs）对大肠杆菌（Escherichia coli，E.coli）毒性作用表征SAs的胁迫作用，用突变和接合转移表征ARGs的产生与传播，测定了SAs对大肠杆菌的毒性、突变频率和结合转移频率的影响，根据剂量-效应曲线，计算了毒性参数（无观察效应浓度（NOEC）、抑制率为50%的化合物浓度（EC50）、抑制率为80%的化合物浓度（EC80）），突变效应参数（促进率为1%时最低可观测突变促进效应浓度（MC0-1）、促进率为50%时突变促进效应浓度（MC50）、促进率最大时突变促进效应浓度（MCmax））和接合转移效应参数（促进率为1%时最低可观测接合转移促进效应浓度（RC0-1）、促进率最大时接合转移促进效应浓度（RCmax）和促进率为1%时最高可观测接合转移促进效应浓度（RC0-2）），利用线性回归分析的方法探究SAs的胁迫与大肠杆菌突变频率和结合转移频率之间的关系，并分析其可能的机制。结果表明，磺胺的高胁迫作用导致核苷酸碱基的大量减少，在DNA复制与转录时，碱基对错配的概率大大增加，从而开始促进突变频率。SAs的低胁迫作用可能引起大肠杆菌的SOS反应，SOS反应可以上调质粒编码的基因以及控制细胞膜的通透性基因，从而提高其接合转移频率。此外，真实环境中存在许多其他的因素也会影响ARGs的产生和传播，据此，本文建议在探索真实环境中ARGs的产生和传播时，应考虑真实环境中其他影响因素和抗生素胁迫的综合作用。上述研究为探索抗生素胁迫对ARGs产生与传播的影响提供了新的思路。Abstract: The pollution induced by antibiotic residues and antibiotic resistance genes (ARGs) has been significantly threating the global public health. Although the relationship between antibiotics and ARGs in the environment have been studied by many investigators, there is no unified conclusion for these results. To investigate the relationship of antibiotics stress with the emergence and dissemination of ARGs, the effects of sulfonamides (SAs) on growth, mutation frequency and conjugative transfer frequency were determined using Escherichia coli (E. coli) as a target organism in this study. The SAs stress on E. coli was expressed as the toxic effects of SAs on the growth of E. coli, and the emergence and dissemination of ARGs were characterized by the mutation and conjugative transfer, respectively. The typical mutation parameters, including minimum observable concentration of SAs (MC0-1), the concentration of SAs that induces the 50% promotion (MC50) and maximum promotion (MCmax) were respectively selected to describe the onset of stimulation, the moderate stimulation, and the maximum stimulation effect of mutation frequency. The typical conjugative transfer parameters, including the minimum observable concentration of SAs (RC0-1), the concentration of SAs that induces the maximum promotion (RCmax), and the maximum observable concentration of SAs (RC0-2) were respectively selected to describe the onset of stimulation, the maximum stimulation, and the terminal stimulation effect of conjugative transfer frequency. The linear regression was conducted to analyze the relationship of the stress of SAs with the mutation frequency and conjugative transfer frequency in E. coli and explore their possible mechanism. The results show that the high stress of SAs results in a substantial reduction in the number of nucleotide bases, which increases the probability of base pair mismatch during DNA replication and transcription, thus promoting the mutation frequency. The low stress of SAs may trigger the SOS response in E. coli, which promotes the conjugative transfer frequency by up-regulating the genes encoded by plasmid and controlling the membrane permeability. Previous studies suggested that many complicated influence factors in the real environment might impact the emergence and dissemination of ARGs, such as the type of compound, concentration of the compound, and transmitting medium. Therefore, it is necessary to comprehensively consider these possible factors when exploring the relationship of antibiotic stress with the emergence and dissemination of ARGs. The present research proposes a new insight into the emergence and dissemination of ARGs under the stress of antibiotics.
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Key words:
- antibiotics /
- sulfonamides /
- antibiotic resistance genes /
- mutation /
- conjugative transfer
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