白屈菜红碱抗耐甲氧西林金黄色葡萄球菌作用机制研究

doi:10.11843/j.issn.0366-6964.2024.10.038

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (10): 4670-4678.doi: 10.11843/j.issn.0366-6964.2024.10.038

白屈菜红碱抗耐甲氧西林金黄色葡萄球菌作用机制研究

苑庆欣¹(), 刘阔¹, 包旭华¹, 高东阳¹, 李鹤¹, 宋军¹^,*(), 周志新²^,*()

1. 黑龙江八一农垦大学动物科技学院农业农村部东北寒区牛病防治重点实验室, 大庆 163319
2. 黑龙江农业经济职业学院动物科技系, 牡丹江 157041

收稿日期:2023-12-08 出版日期:2024-10-23 发布日期:2024-11-04
通讯作者: 宋军,周志新 E-mail:yuanqingxin_2000@126.com;songjun_2005@126.com;Z2855877761@126.com
作者简介:苑庆欣(2000-), 男, 黑龙江青冈人, 硕士生, 主要从事动物性/食源性致病菌防控研究, E-mail: yuanqingxin_2000@126.com
基金资助:
国家自然科学基金青年科学基金项目(31802226);黑龙江省自然科学基金联合引导项目(LH2022C072);黑龙江省牛病防制重点实验室开放课题(PCBD201710)

The Mechanism of Chelerythrine against Methicillin-resistant Staphylococcus aureus

Qingxin YUAN¹(), Kuo LIU¹, Xuhua BAO¹, Dongyang GAO¹, He LI¹, Jun SONG¹^,*(), Zhixin ZHOU²^,*()

1. Key Laboratory of Bovine Disease Control in Northeast China of Ministry of Agriculture and Rural Affairs, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China
2. Department of Animal Science and Technology, Heilongjiang Agricultural Economy Vocational College, Mudanjiang 157041, China

Received:2023-12-08 Online:2024-10-23 Published:2024-11-04
Contact: Jun SONG, Zhixin ZHOU E-mail:yuanqingxin_2000@126.com;songjun_2005@126.com;Z2855877761@126.com

摘要/Abstract

摘要：

本研究旨在阐明白屈菜红碱(chelerythrine, CHE)抗耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus, MRSA)活性及探究其抗菌机制。通过测定最小抑菌浓度及最小杀菌浓度分析CHE对MRSA的抗菌活性；检测CHE对MRSA细胞壁及细胞膜的完整性、通透性，结合电镜观察MRSA细胞形态和超微结构，从而对CHE抗菌机制进行初步分析；检测CHE对MRSA菌株活性氧(reactive oxygen species, ROS)水平与ROS相关因子造成的影响，并结合荧光定量及转录组学技术进一步验证CHE抗菌机制。结果表明，CHE对MRSA的最小抑菌浓度为6.25 μg·mL^-1，最小杀菌浓度为12.5 μg·mL^-1。CHE可通过破坏MRSA菌株细胞细胞壁及细胞膜，发挥抗菌作用。且CHE处理后可抑制MRSA呼吸链SdhABC等基因表达，从而阻断呼吸链电子传递，使ROS水平升高从而发挥抗菌作用。综上，本研究对CHE抗MRSA的抗菌机制进行探究，为临床防治MRSA感染提供新参考。

关键词: 耐甲氧西林金黄色葡萄球菌, 白屈菜红碱, 抗菌机制, 氧化应激

Abstract:

The objective of this study was to investigate the antibacterial effect of chelerythrine (CHE) against methicillin-resistant Staphylococcus aureus (MRSA) and approach its mechanism. The antibacterial activity of CHE against MRSA was determined by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). After treatment with CHE, the cell membrane and permeability of MRSA were assessed, and the morphology and ultrastructure of MRSA cells were observed by electron microscopy to preliminarily analyze the antibacterial mechanism of CHE. Additionally, the effect of CHE on reactive oxygen species (ROS) levels and ROS-related factors in MRSA strains was detected, combined with fluorescence quantification and transcriptomics to further explore the antibacterial mechanism of CHE. The results showed that the MIC of CHE against MRSA was 6.25 μg·mL^-1, MBC is 12.5 μg·mL^-1, and CHE can destroy cell membrane integrity and permeability to achieve antibacterial effect. CHE may block the electronic transmission of respiratory chain and promote ROS production by inhibiting the expression of gene SdhABC and other genes to exert activity against MRSA. In conclusion, this study explored the antibacterial mechanism of CHE against MRSA, which provided a new reference for clinical prevention of MRSA infection.

Key words: MRSA, chelerythrine, antibacterial effect, oxidative stress

中图分类号:

S853.74

苑庆欣, 刘阔, 包旭华, 高东阳, 李鹤, 宋军, 周志新. 白屈菜红碱抗耐甲氧西林金黄色葡萄球菌作用机制研究[J]. 畜牧兽医学报, 2024, 55(10): 4670-4678.

Qingxin YUAN, Kuo LIU, Xuhua BAO, Dongyang GAO, He LI, Jun SONG, Zhixin ZHOU. The Mechanism of Chelerythrine against Methicillin-resistant Staphylococcus aureus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4670-4678.

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基因Gene	序列(5'→3') Sequences	产物长度/bp Product
gene_01103	F: ACGGCAAAGAAGTGAATTACGA R: GCAAAGTGGAACACAACAGC	103
gene_01104	F: ACCGCTTAGGTGCCAATTCA R: GCATTTTCTGTACCGCGCAT	190

基因ID Gene ID	产物名称Production name	差异表达倍数Fold change
gene_01103	琥珀酸脱氢酶细胞色素b-558亚基(SdhC)	-1.32
	Succinate dehydrogenase cytochrome b-558 subunit
gene_01104	琥珀酸脱氢酶黄蛋白亚单位(SdhA)	-1.73
	Succinate dehydrogenase flavoprotein subunit

通路Pathway	一级分类Level 1	上调Up	下调Down
核糖体 Ribosome	遗传信息处理 Genetic information processing	0	49
光合作用Photosynthesis	新陈代谢Metabolism	0	8
精氨酸生物合成Arginine biosynthesis	新陈代谢Metabolism	6	8
半乳糖代谢Galactose metabolism	新陈代谢Metabolism	14	0
RNA聚合酶 RNA polymerase	遗传信息处理 Genetic information processing	0	5
细菌分泌系统 Bacterial secretion system	环境信息处理 Environmental information processing	4	6
双组分系统 Two-component system	环境信息处理 Environmental information processing	21	22
阳离子抗菌肽(CAMP)耐药性 Cationic antimicrobial peptide (CAMP) resistance	人类疾病 Human diseases	3	6
生物素代谢Biotin metabolism	新陈代谢Metabolism	5	3
O-抗原核苷酸糖生物合成 O-antigen nucleotide sugar biosynthesis	新陈代谢 Metabolism	7	2
阿特拉津降解Atrazine degradation	新陈代谢Metabolism	3	0
各种次生代谢产物的生物合成—第3部分 Biosynthesis of various secondary metabolites-part 3	新陈代谢 Metabolism	2	1
C5支链二元酸代谢 C5-branched dibasic acid metabolism	新陈代谢 Metabolism	0	7
ABC转运蛋白 ABC transporters	环境信息处理 Environmental information processing	28	16
蛋白质外排 Protein export	遗传信息处理 Genetic information processing	4	6
氧化磷酸化Oxidative phosphorylation	新陈代谢Metabolism	0	13
β内酰胺耐药性β-Lactam resistance	人类疾病Human diseases	0	6
幽门螺杆菌感染中的上皮细胞信号传导 Epithelial cell signaling in helicobacter pylori infection	人类疾病 Human diseases	1	1
植物-病原体相互作用Plant-pathogen interaction	生物系统Organismal systems	0	2
群体感应Quorum sensing	细胞过程Cellular processes	22	11

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白屈菜红碱抗耐甲氧西林金黄色葡萄球菌作用机制研究

The Mechanism of Chelerythrine against Methicillin-resistant Staphylococcus aureus

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