He Liren, Cheng Han, Chen Fuxin, Song Suquan, Zhang Hang, Sun Weidong, Bao Xiaowei, Zhang Haibin, He Chenghua
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
School of Chemistry and Chemical Engineering, Xi'an University of Science and Technology, Xi'an 710054, China.
Vet Sci. 2022 Feb 7;9(2):71. doi: 10.3390/vetsci9020071.
(1) Background: Methicillin-resistant (MRSA) is a zoonotic pathogen that causes endocarditis, pneumonia, and skin diseases in humans and livestock. (2) Methods: The antibacterial effect of the total flavonoid against MRSA (ATCC43300) extracted from the Ledeb. ( Ledeb) was evaluated by the microdilution method. The oxidative stresses in MRSA were evaluated by the levels of intracellular hydrogen peroxide (HO), reactive oxygen species (ROS), and oxidative stress-related genes. The DNA oxidative damage was tested by the 8-hydroxy-2'-deoxyguanosine (8-OHdG) and DNA gel electrophoresis. The differentially expressed proteins were determined by the method of SDS-PAGE and NanoLC-ESI-MS/MS, while the mRNAs of differential proteins were determined by Real-Time PCR. The changes of ultra-structures in MRSA were observed by Transmission Electron Microscope (TEM). (3) Results: The minimum inhibitory concentration (MIC) of the total flavonoid against MRSA was recorded as 62.5 μg/mL. After treatment with the total flavonoid, the levels of intracellular HO and ROS were increased and the gene expressions against oxidative stress (, , ) were decreased ( < 0.01), while the gene expression for oxidative stress () was increased ( < 0.01). The level of intracellular 8-OHdG in MRSA was increased ( < 0.01) and the DNA was damaged. The results of TEM also showed that the total flavonoid could destroy the ultra-structures in the bacteria. (4) Conclusions: The total flavonoid extracted from the Ledeb can induce the oxidative stress that disturbed the energy metabolism and protein synthesis in MRSA.
(1)背景:耐甲氧西林金黄色葡萄球菌(MRSA)是一种人畜共患病原体,可导致人类和牲畜患心内膜炎、肺炎和皮肤病。(2)方法:采用微量稀释法评估从绵毛酸模叶蓼(绵毛酸模叶蓼)中提取的总黄酮对MRSA(ATCC43300)的抗菌效果。通过细胞内过氧化氢(HO)、活性氧(ROS)水平以及氧化应激相关基因评估MRSA中的氧化应激。通过8-羟基-2'-脱氧鸟苷(8-OHdG)和DNA凝胶电泳检测DNA氧化损伤。采用SDS-PAGE和NanoLC-ESI-MS/MS方法测定差异表达蛋白,同时通过实时PCR测定差异蛋白的mRNA。通过透射电子显微镜(TEM)观察MRSA中超微结构的变化。(3)结果:总黄酮对MRSA的最低抑菌浓度(MIC)记录为62.5μg/mL。用总黄酮处理后,细胞内HO和ROS水平升高,抗氧化应激基因(,,)的表达降低(<0.01),而抗氧化应激基因()的表达增加(<0.01)。MRSA细胞内8-OHdG水平升高(<0.01),DNA受到损伤。TEM结果还表明,总黄酮可破坏细菌中的超微结构。(4)结论:从绵毛酸模叶蓼中提取的总黄酮可诱导氧化应激,扰乱MRSA中的能量代谢和蛋白质合成。