秀丽金老鹳草总黄酮抑制耐甲氧西林金黄色葡萄球菌（MRSA）生物膜形成和毒力因子的产生。
Total flavonoids from Potentilla kleiniana Wight et Arn inhibits biofilm formation and virulence factors production in methicillin-resistant Staphylococcus aureus (MRSA).
机构信息
College of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen, 518000, China; Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, 518000, China; School of Food Engineering and Biotechnology, Hanshan Normal University, Chaozhou, 521041, China.
College of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen, 518000, China.
出版信息
J Ethnopharmacol. 2021 Oct 28;279:114383. doi: 10.1016/j.jep.2021.114383. Epub 2021 Jun 29.
ETHANOPHARMACOLOGICAL RELEVANCE
Potentilla kleiniana Wight et Arn is a wide-spread wild plant in the mountainous areas in southern China. The whole herb has been used as a traditional herbal medicine to treat fever, arthritis, malaria, insect and snake bites, hepatitis, and traumatic injury. In vitro studies have reported the antibacterial activity use of the plant in traditional medicinal systems.
AIM OF THE STUDY
The aim of this study was to investigate the inhibitory activity of total flavonoid from Potentilla kleiniana Wight et Arn (TFP) on methicillin-resistant Staphylococcus aureus (MRSA) in planktonic state and biofilm state.
MATERIALS AND METHODS
Antibacterial activities of TFP on planktonic MRSA were determined by agar diffusion method, microtiter plate assay and time-kill curve assay. Electrical conductivity, membrane permeability, membrane potential and autoaggregation were analyzed to study TFP effects on planktonic MRSA growth. Crystal violet (CV) staining and confocal laser scanning microscopy (CLSM) were analyzed to study TFP effects on aggregation and maturation of MRSA biofilm. After TFP treatment, extracellular polymeric substances (EPS) production were examined. Morphological changes in planktonic and MRSA biofilm following TFP treatment were determined with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Moreover, α-Toxin protein expression and adhesion-related gene expression were also determined.
RESULTS
The minimum inhibitory concentration (MIC) of TFP against MRSA was 20 μg/mL. The agar diffusion method and time-kill curve assay results indicated that TFP inhibited planktonic MRSA growth. TFP treatment significantly inhibited planktonic MRSA growth by inhibiting autoaggregation, α-hemolysin activity, α-Toxin protein expression, but increasing electrolyte leakage, membrane permeability and membrane potential and impacting cell structure. Moreover, TFP treatment significantly inhibited aggregation and maturation on MRSA biofilm by decreasing surface hydrophobicity, EPS production and adhesion-related gene expression.
CONCLUSION
The results of this trial provide scientific experimental data on the traditional use of Potentilla Kleiniana Wight et Arn for traumatic injury treatment and further demonstrate the potential of TFP to be developed as a novel anti-biofilm drug.
ethanopharmacological 相关性：翻白草是中国南方山区广泛分布的野生植物。全草被用作传统草药，用于治疗发热、关节炎、疟疾、昆虫和蛇咬伤、肝炎和创伤性损伤。体外研究报告了该植物在传统药用系统中的抗菌活性。
研究目的
本研究旨在探讨翻白草总黄酮（TFP）对浮游状态和生物膜状态下耐甲氧西林金黄色葡萄球菌（MRSA）的抑制活性。
材料和方法
采用琼脂扩散法、微量板法和时间杀伤曲线法测定 TFP 对浮游 MRSA 的抗菌活性。分析电导率、膜通透性、膜电位和自动聚集，以研究 TFP 对浮游 MRSA 生长的影响。结晶紫（CV）染色和共聚焦激光扫描显微镜（CLSM）分析用于研究 TFP 对 MRSA 生物膜聚集和成熟的影响。TFP 处理后，检测细胞外聚合物物质（EPS）的产生。通过扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察 TFP 处理后浮游和 MRSA 生物膜的形态变化。此外，还测定了α-毒素蛋白表达和粘附相关基因表达。
结果
TFP 对 MRSA 的最小抑菌浓度（MIC）为 20μg/ml。琼脂扩散法和时间杀伤曲线法结果表明，TFP 抑制浮游 MRSA 生长。TFP 处理通过抑制自动聚集、α-溶血素活性、α-毒素蛋白表达，增加电解质泄漏、膜通透性和膜电位，影响细胞结构，显著抑制浮游 MRSA 的生长。此外，TFP 处理通过降低表面疏水性、EPS 产生和粘附相关基因表达，显著抑制 MRSA 生物膜的聚集和成熟。
结论
本试验结果为翻白草传统用于创伤治疗提供了科学实验数据，并进一步证明了 TFP 作为新型抗生物膜药物的开发潜力。
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