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氧化锌纳米棒通过抑制青霉素结合蛋白2a对耐甲氧西林菌的抑制机制

Inhibition Mechanism of Methicillin-Resistant by Zinc Oxide Nanorods via Suppresses Penicillin-Binding Protein 2a.

作者信息

Hassan Amr, Al-Salmi Fawziah A, Saleh Muneera A, Sabatier Jean-Marc, Alatawi Fuad A, Alenezi Muneefah Abdullah, Albalwe Fauzeya M, Meteq R Albalawi Hessa, Darwish Doaa Bahaa Eldin, Sharaf Eman M

机构信息

Department of Bioinformatics, Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Sadat 32897, Egypt.

Department of Biology, Faculty of Sciences, Taif University, Taif 21944, Saudi Arabia.

出版信息

ACS Omega. 2023 Mar 6;8(11):9969-9977. doi: 10.1021/acsomega.2c07142. eCollection 2023 Mar 21.

Abstract

Methicillin-resistant (MRSA) causes life-threatening infections. Zinc oxide is well known as an effective antibacterial drug against many bacterial strains. We investigated the performance of zinc oxide nanorods synthesized by Albmiun as a biotemplate as an antibacterial drug in this study; the fabrication of zinc oxide nanorods was synthesized by sol-gel methods. We performed physicochemical characterization of zinc oxide nanorods by physiochemical techniques such as FTIR spectroscopy, X-ray diffraction, and TEM and investigation of their antimicrobial toxicity efficiency by MIC, ATPase activity assay, anti-biofilm activity, and kill time assays, as well as the mecA, mecR1, blaR1, blaZ, and biofilm genes (ica A, ica D, and fnb A) by using a quantitative RT-PCR assay and the penicillin-binding protein 2a (PBP2a) level of MRSA by using a Western blot. The data confirmed the fabrication of rod-shaped zinc oxide nanorods with a diameter in the range of 50 nm, which emphasized the formation of zinc oxide nanoparticles with regular shapes. The results show that zinc oxide nanorods inhibited methicillin-resistant effectively. The MIC value was 23 μg/mL. The time kill of ZnO-NRs against MRSA was achieved after 2 h of incubation at 4MIC (92 μg/mL) and after 3 h of incubation at 2MIC (46 μg/mL), respectively. The lowest concentration of zinc oxide nanorods with over 75% biofilm killing in all strains tested was 32 μg/mL. Also, we examined the influence of the zinc oxide nanorods on MRSA by analyzing mecA, mecR1, blaR1, and blaZ by using a quantitative RT-PCR assay. The data obtained revealed that the presence of 2× MIC (46 μg/mL) of ZnO-NRs reduced the transcriptional levels of blaZ, blaR1, mecA, and mecR1 by 3.4-fold, 3.6-fold, 4-fold, and 3.8-fold, respectively. Furthermore, the gene expression of biofilm encoding genes (ica A, ica B, ica D, and fnb A) was tested using quantitative real-time reverse transcriptase-polymerase chain reaction (rt-PCR). The results showed that the presence of 2× MIC (46 μg/mL) of ZnO-NRs reduced the transcriptional levels of ica A, ica B, ica D, and fnb A. Also, the PBP2a level was markedly reduced after treatment with ZnO-NRs.

摘要

耐甲氧西林金黄色葡萄球菌(MRSA)可引发危及生命的感染。氧化锌作为一种对多种细菌菌株有效的抗菌药物广为人知。在本研究中,我们调查了以白蛋白作为生物模板合成的氧化锌纳米棒作为抗菌药物的性能;氧化锌纳米棒是通过溶胶 - 凝胶法合成的。我们通过傅里叶变换红外光谱(FTIR)、X射线衍射和透射电子显微镜(TEM)等物理化学技术对氧化锌纳米棒进行了物理化学表征,并通过最低抑菌浓度(MIC)、ATP酶活性测定、抗生物膜活性和杀菌时间测定来研究其抗菌毒性效率,同时通过定量逆转录聚合酶链反应(RT - PCR)测定mecA、mecR1、blaR1、blaZ和生物膜基因(ica A、ica D和fnb A),并通过蛋白质印迹法测定MRSA的青霉素结合蛋白2a(PBP2a)水平。数据证实了直径在50nm范围内的棒状氧化锌纳米棒的制备,这强调了具有规则形状的氧化锌纳米颗粒的形成。结果表明,氧化锌纳米棒能有效抑制耐甲氧西林金黄色葡萄球菌。MIC值为23μg/mL。氧化锌纳米棒(ZnO - NRs)对MRSA的杀菌时间分别在4MIC(92μg/mL)孵育2小时后和2MIC(46μg/mL)孵育3小时后达到。在所有测试菌株中,具有超过75%生物膜杀灭率的氧化锌纳米棒的最低浓度为32μg/mL。此外,我们通过定量RT - PCR测定mecA、mecR1、blaR1和blaZ来研究氧化锌纳米棒对MRSA的影响。获得的数据显示,2×MIC(46μg/mL)的ZnO - NRs的存在分别使blaZ、blaR1、mecA和mecR1的转录水平降低了3.4倍、3.6倍、4倍和3.8倍。此外,使用定量实时逆转录聚合酶链反应(rt - PCR)检测生物膜编码基因(ica A、ica B、ica D和fnb A)的基因表达。结果表明,2×MIC(46μg/mL)的ZnO - NRs的存在降低了ica A、ica B、ica D和fnb A的转录水平。而且,用ZnO - NRs处理后,PBP2a水平明显降低。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8a9a/10034842/ca8d66bd34f4/ao2c07142_0002.jpg

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