首页|巴西苏木素对耐甲氧西林金黄色葡萄球菌葡萄黄质的抑制作用及其机制

巴西苏木素对耐甲氧西林金黄色葡萄球菌葡萄黄质的抑制作用及其机制

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目的 探究巴西苏木素对耐甲氧西林金黄色葡萄球菌(MRSA)葡萄黄质(STX)的抑制作用及其机制。方法 基于转录组学数据，分析巴西苏木素作用于MRSA后编码STX合成的操纵子crtOPQMN基因变化情况;将不同浓度巴西苏木素(16、32、64 μg/ml)与MRSA共培养:转平板观察菌落、离心沉淀观察菌体颜色变化及酶标仪检测STX含量;逆转录实时定量聚合酶链反应(qRT-PCR)检测STX合成中主要影响颜色的基因表达水平变化;光谱定量检测STX代谢中间产物变化情况;过氧化氢敏感性试验观察抑菌圈大小及存活菌数量;全血杀伤试验观察存活菌数量。结果 调控STX生成的操纵子基因表达均下调，其中主要影响颜色的基因crtM和crtN分别为-6。542479954、-6。107726632;随药物浓度增加，平板菌落及菌沉淀黄色变浅变白，STX含量减少，其中64 μg/ml药物对STX抑制率最大，达50％;qRT-PCR结果显示随药物浓度增加crtM和crtN表达量下降;光谱定量结果显示STX代谢中间产物含量随药物浓度增加而减少;过氧化氢敏感性试验结果显示随药物浓度增大，抑菌圈直径增大，存活菌数量减低(P＜0。05);全血杀伤试验结果显示随药物浓度增大，存活菌数量减低(P＜0。05)。结论 巴西苏木素通过抑制crtM和crtN的表达减少MRSA STX生成;巴西苏木素抑制STX后可降低细菌抗氧化及抗宿主免疫杀伤能力，为开发新型抗毒力药物提供参考。
Inhibitory effect of Brazilin on staphyloxanthin of methicillin-resistant Staphylococcus aureus and its mechanisms
OBJECTIVE To explore the inhibitory effect of Brazilin on staphyloxanthin(STX)of methicillin-resist-ant Staphylococcus aureus(MRSA)and analyze its mechanisms.METHODS Based on transcriptomic data,the change of operon crtOPQMN gene synthesized from encoding STX was observed after Brazilin acted on MRSA.The different concentrations of Brazilin(16,32,64 μg/ml)were cocultured with MRSA;the bacterial colonies were observed by plate transfer,the change of color of bacteria was observed by centrifugation precipitation,and the content of STX was detected by microplate reader.The changes of expression levels of STX encoding genes that mainly affected the colors were detected by means of reverse transcription real-time quantitative PCR(qRT-PCR),the changes of STX metabolic intermediate products were quantitatively detected by spectra,the size of in-hibition circle and colony counts of surviving bacteria were observed by hydrogen peroxide sensitivity assay,and the bacterial colony counts of surviving bacteria were observed by means of whole blood killing assay.RESULTS The expressions of operon genes regulating the production of STX were down-regulated;crtM and crtN genes that mainly affected the color were-6.542479954 and-6.107726632.As the concentration of drug increases,the yellow color of bacterial colonies on plate and precipitates became lighter and white,the content of STX was re-duced,the inhibition rate of the 64 μg/ml of drug on STX was greatest and reached up to 50％.The result of qRT-PCR showed that the expression levels of crtM and crtN declined with the increase of drug concentration.The re-sult of spectra quantitative assay indicated that the content of STX metabolic intermediate products was reduced with the increase of the drug concentration.The result of hydrogen peroxide sensitivity assay showed that with the increase of drug concentration,the diameter of inhibition circle was increased,and the colony counts of surviving bacteria was significantly reduced(P＜0.05).The result of whole blood killing assay showed that the colony counts of surviving bacteria were reduced significantly with the increase of drug concentration(P＜0.05).CONCLUSION Brazilin can reduce the production of MRSA STX by inhibiting the expressions of crtM and crtN.It can reduce the antioxidant and anti-host immune killing ability of bacteria after the inhibition of STX,which may provide reference for development of novel antitoxic drugs.

BrazilinMethicillin-resistant Staphylococcus aureusStaphyloxanthincrtM genecrtN geneAn-tioxidantAnti-killing

周永雯、陈泽慧、周小仙、杨智芳、余孟飞、刘和兰、黎瑞、李明哲

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遵义医科大学附属医院医学检验科,贵州遵义 563003

遵义医科大学检验医学院,贵州遵义 563006

遵义医科大学第二附属医院医学检验科,贵州遵义 563000

遵义医科大学附属医院输血科,贵州遵义 563003

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巴西苏木素 耐甲氧西林金黄色葡萄球菌 葡萄黄质 crtM基因 crtN基因 抗氧化 抗杀伤

贵州省科学技术厅科学技术基金遵义市科技计划遵义市科技计划

黔科合支撑[2021]一般034黔科平台人才[2020]-025遵市科合HZ字[2021]299号

2024

10.11816/cn.ni.2024-231154

中华医院感染学杂志
中华预防医学会 中国人民解放军总医院

中华医院感染学杂志

CSTPCD北大核心
影响因子：1.885
ISSN：1005-4529
年,卷(期)：2024.34(5)
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