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单宁酸对耐甲氧西林金黄色葡萄球菌生物膜抑制作用的蛋白质组学研究

Proteomic study of the inhibitory effects of tannic acid on MRSA biofilm.

作者信息

Miao Yang, Shuang Wang, Qianwei Qu, Xin Liu, Wei Peng, Hai Yang, Yonghui Zhou, Xinbo Yu

机构信息

School of Basic Medicine, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, China.

College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, China.

出版信息

Front Pharmacol. 2024 Dec 18;15:1413669. doi: 10.3389/fphar.2024.1413669. eCollection 2024.

Abstract

INTRODUCTION

The mechanism of tannic acid (TA) intervention on methicillin-resistant (MRSA, USA 300) biofilm formation was explored using proteomics.

METHODS

The minimum inhibitory concentration (MIC) of TA against the MRSA standard strain USA 300 was determined by two-fold serial dilution of the microbroth. The effects of TA were studied using crystal violet staining. The morphology of TA-treated USA 300 cells was observed by scanning electron microscopy and confocal laser scanning microscopy. Differentially expressed proteins (DEPs) were screened using proteomic and biological information analyses, and their transcriptional levels were verified using real-time quantitative polymerase chain reaction.

RESULTS

The MIC of TA was 0.625 mg/mL, whereas 1/2 MIC (0.3125 mg/mL) of TA significantly inhibited biofilm formation without affecting the bacterial growth ( < 0.01) and prevented the formation of a complete three-dimensional biofilm structure. Using 1/2 MIC of TA, 208 DEPs were identified, of which 127 were upregulated and 81 were downregulated. The transcriptional levels of the genes corresponding to five randomly selected DEPs ( and ) were consistent with the proteomics data ( < 0.05). Bioinformatic analysis showed that the changes in the MRSA strains after TA intervention primarily involved pyrimidine and purine metabolisms, arginine biosynthesis, and the citric acid cycle.

CONCLUSION

TA exerts an antibacterial effect on MRSA and can be used as a potential candidate for the development of anti-biofilm drugs, thereby laying a foundation for the treatment of MRSA biofilm-induced infections.

摘要

引言

采用蛋白质组学方法探讨了鞣酸(TA)干预耐甲氧西林金黄色葡萄球菌(MRSA,美国300型)生物膜形成的机制。

方法

采用微量肉汤二倍稀释法测定TA对MRSA标准菌株美国300型的最低抑菌浓度(MIC)。采用结晶紫染色法研究TA的作用效果。通过扫描电子显微镜和共聚焦激光扫描显微镜观察TA处理后美国300型细胞的形态。利用蛋白质组学和生物信息学分析筛选差异表达蛋白(DEP),并通过实时定量聚合酶链反应验证其转录水平。

结果

TA的MIC为0.625mg/mL,而1/2 MIC(0.3125mg/mL)的TA能显著抑制生物膜形成,且不影响细菌生长(P<0.01),并阻止完整三维生物膜结构的形成。使用1/2 MIC的TA,鉴定出208个DEP,其中127个上调,81个下调。随机选择的5个DEP(……和……)对应的基因转录水平与蛋白质组学数据一致(P<0.05)。生物信息学分析表明,TA干预后MRSA菌株的变化主要涉及嘧啶和嘌呤代谢、精氨酸生物合成以及柠檬酸循环有关。

结论

TA对MRSA具有抗菌作用,可作为开发抗生物膜药物的潜在候选药物,为治疗MRSA生物膜引起的感染奠定基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebee/11688184/7416b67a709c/fphar-15-1413669-g001.jpg

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