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| 噬菌体裂解酶TSPphg对金黄色葡萄球菌生物被膜的抑制作用及该过程转录组变化 |
| Inhibition of Staphylococcus aureus Biofilm by Phage Lyase TSPphg and Transcriptome Changes of The Process |
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| DOI：doi:10.3969/j.issn.1005-7021.2025.01.003 |
| 中文关键词: 生物被膜 金黄色葡萄球菌 噬菌体裂解酶 加权共表达网络分析（WGCNA） 代谢通路 核心驱动因子 |
| 英文关键词: biofilm Staphylococcus aureus phage lyase weighted co expression network analysis (WGCNA) metabolic pathway core driver |
| 基金项目:国家自然科学基金项目（32260040）；兴滇英才支持计划-青年人才专项（YNWR-QNBJ-2020-087） |
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| 中文摘要: |
| 研究噬菌体裂解酶对细菌生物被膜(Bacterial biofilm, BBF)的作用效果及转录机制。细菌生物被膜的形成是细菌耐药性产生的重要原因之一。金黄色葡萄球菌(Staphylococcus aureus)是常见的易形成生物被膜的致病微生物，生物被膜的形成给临床细菌性感染的治疗和医疗设备表面细菌的清除带来很大的困难。使用结晶紫染色法、XTT(2,3-BIS (2-methoxy-4-nitro-5-sulfonyl)-2H-tetrazolium-5-carboxanilide)实验、荧光显微镜(Fluorescence Microscope,FM)及扫描电子显微镜(Scanning Electron Microscope,SEM)观察等方法研究噬菌体裂解酶TSPphg对金黄色葡萄球菌生物被膜的抑制效果，利用转录组测序得到噬菌体裂解酶TSPphg降解和抑制金黄色葡萄球菌生物被膜过程中的差异基因，并采用加权共表达网络分析(Weighted Correlation Network Analysis, WGCNA)获取其关键模块，进行GO功能注释，并构建其调控因子网络。纯化获得噬菌体裂解酶TSPphg重组蛋白，实验中发现噬菌体裂解酶TSPphg对金黄色葡萄球菌生物被膜形成具有良好的抑制作用，且显微镜下观察到作用后的生物被膜密度显著变小，说明其能达到清除细菌生物被膜的效果。进而通过转录组和WGCNA分析揭示了其中的关键调控模块及分子网络。裂解酶TSPphg显示了良好的金黄色葡萄球菌生物被膜抑制效果，解析出了该过程中的差异基因、特征性调控模块及核心因子网络，为抗生素替代、生物被膜的清除提供了新的思路。 |
| 英文摘要: |
| Bacterial biofilm (BBF) formation is one of the important causes of bacterial drug resistance. Staphylococcus aureus is a common pathogenic microorganism that is prone to biofilm formation, which makes the treatment of bacterial infections and the removal of bacteria from the surface of medical devices clinically very difficult. To investigate the effect of phage lyase on the bacterial biofilm and the transcriptional mechanism. The effect of phage lyase TSPphg on the biofilm inhibition of S. aureus was investigated by crystalline violet staining, XTT (2,3-BIS (2-methoxy-4-nitro-5-sulfonyl)-2H-tetrazolium-5-carboxanilide) assay, fluorescence microscope (FM) and scanning electron microscope (SEM) observation to study the effect of phage lyase TSPphg on S. aureus biofilm inhibition. Transcriptome sequencing was used to obtain the differential genes in the process of phage lyase TSPphg treatment of S. aureus biofilm, and used weighted co-expression network analysis (WGCNA) to obtain its key modules, perform GO functional annotation, and subsequently construct the network of its core factors. As a result, the purified protein of phage lytic enzyme TSPphg was obtained, and it was found that phage lyase TSPphg had a good inhibitory effect on the biofilm formation of S. aureus and the density of biofilm was significantly smaller after the action was observed under microscope, indicating that it could achieve the effect of removing bacterial biofilm. Further transcriptome and WGCNA analyses of key genes in the degradation and inhibition processes were performed. The lyase TSPphg showed good biofilm inhibition against S. aureus, and parsed the differential genes, characteristic regulatory modules and core factor networks in this process were resolved, and this study provided train of thought to purge the biofilm for the replacement of antibiotics. |
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