吡唑啉酮铜配合物P-FAH-Cu-phen对金黄色葡萄球菌的转录组影响分析

Pyrazolone-copper complex P-FAH-Cu-phen affects transcriptome of Staphylococcus aureus

摘要:

【背景】金黄色葡萄球菌是目前食品和临床引起感染的重要病原菌，迫切需要开发新型抗菌药物。【目的】分析吡唑啉酮铜配合物P-FAH-Cu-phen对金黄色葡萄球菌的转录组影响和主要代谢信号通路。【方法】采用液体稀释法测定P-FAH-Cu-phen作用金黄色葡萄球菌的最低抑菌浓度(minimum inhibitory concentration, MIC)和最低杀菌浓度(minimum bactericidal concentration, MBC)。将终浓度2 μg/mL的配合物分别作用于对数生长期的金黄色葡萄球菌30 min和2 h，进行转录组测序及分析。【结果】 P-FAH-Cu-phen作用金黄色葡萄球菌的MIC和MBC分别为2 μg/mL和4 μg/mL。与空白对照相比，配合物处理细菌30 min后，其差异基因共有356个，其中上调表达180个、下调表达176个；配合物处理细菌2 h后，其差异基因共有23个，其中上调表达3个、下调表达20个。差异基因功能主要富集于膜的组成部分、细胞质、质膜、ATP结合、发病机制、金属离子结合、组氨酸生物合成过程、DNA结合、水解酶活性、跨膜转运蛋白活性、硝酸盐同化、硝酸盐代谢过程、硝酸还原酶复合物、硝酸还原酶活性等。差异基因涉及的信号通路主要有双组分系统、群体感应、氮代谢、三羧酸循环、氨基酸代谢等。【结论】影响细菌质膜组成、毒素生成、生物膜形成、细胞壁合成、能量代谢等可能是吡唑啉酮铜配合物P-FAH-Cu-phen对金黄色葡萄球菌的主要抑菌作用。研究为揭示吡唑啉酮铜配合物抑制金黄色葡萄球菌分子机制提供了理论依据。

Abstract:

[Background] Staphylococcus aureus is a major bacterial pathogen causing infections in food and human body, and there is an urgent need to develop new antibacterial agents. [Objective] To study the effects of the pyrazolone-copper complex P-FAH-Cu-phen on the transcriptome and main metabolic signaling pathways of S. aureus. [Methods] The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of P-FAH-Cu-phen on S. aureus were determined by liquid dilution method. The S. aureus in the logarithmic growth phase was treated with P-FAH-Cu-phen at the final concentration of 2 μg/mL for 30 min and 2 h, respectively, and then the transcriptome was sequenced and analyzed. [Results] The MIC and MBC of P-FAH-Cu-phen on S. aureus were 2 μg/mL and 4 μg/mL, respectively. Compared with the blank control, P-FAH-Cu-phen treatment for 30 min led to 356 differentially expressed genes (DEGs), including 180 up-regulated genes and 176 down-regulated genes. P-FAH-Cu-phen treatment for 2 h resulted in 23 DEGs, including 3 up-regulated genes and 20 down-regulated genes. The DEGs were mainly enriched in membrane components, cytoplasm, plasma membrane, ATP binding, pathogenesis, metal ion binding, histidine biosynthesis process, DNA binding, hydrolytic enzyme activity, transmembrane transporter activity, nitrate assimilation, nitrate metabolism process, nitrate reductase complex, nitrate reductase activity, etc. The signaling pathways involving the DEGs mainly included two-component system, quorum sensing, nitrogen metabolism, tricarboxylic acid cycle, amino acid metabolism and so on. [Conclusion] P-FAH-Cu-phen may inhibit S. aureus by affecting plasma membrane composition, toxin production, biofilm formation, cell wall synthesis, and energy metabolism. The study provides a theoretical basis for revealing the molecular mechanism of the inhibition on S. aureus by P-FAH-Cu-phen.