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金银花黄酮脂质体膜的制备及其抗MRSA的机制研究
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| 篇名： | 金银花黄酮脂质体膜的制备及其抗MRSA的机制研究 |
| TITLE： | Preparation of Lonicerae japonicae flavonoids liposome membrane and its antibacterial mechanism against methicillin-resistant Staphylococcus aureus |
| 摘要： | 目的 制备金银花黄酮脂质体膜（LFLM），并研究其抗耐甲氧西林金黄色葡萄球菌（MRSA）的作用机制。方法以大豆磷脂、胆固醇为载体，利用乙醇注入法制备金银花黄酮脂质体，然后将其负载于壳聚糖和聚乙烯醇的混合膜材中制得LFLM。测定金银花黄酮脂质体粒径、多分散性指数（PDI）和LFLM的释放率。设置对照组、LFLM低浓度组（2.5mg/mL）、LFLM中浓度组（5mg/mL）、LFLM高浓度组（10mg/mL）、阳性组（10μg/mL万古霉素）和联合用药组（10mg/mLLFLM+10μg/mL万古霉素），评价LFLM对MRSA菌落形成、存活、生物膜形成的影响；检测MRSA上清液中K+、Mg2+、乳酸脱氢酶（LDH）、碱性磷酸酶（AKP）的含量；检测MRSA中mecA、mecR1mRNA的表达水平。结果金银花黄酮脂质体的粒径为（80.91±3.96）nm，PDI为0.26±0.07；LFLM在12h内的释放率为55%，36h内的释放率为73%。与对照组比较，LFLM中、高浓度组和阳性组MRSA的菌落形成数均显著减少（P＜0.05），死菌与活菌之比、生物膜抑制率以及上清液中K+、Mg2+、LDH、AKP含量均显著升高（P＜0.05），mecA、mecR1mRNA表达水平均显著降低（P＜0.05）。LFLM与万古霉素联用后，抗MRSA作用进一步加强。结论本研究成功制得LFLM，其具有良好的释药性，可通过促进MRSA细胞壁和细胞膜破坏，抑制mecA、mecR1mRNA表达，从而发挥抗菌作用。 |
| ABSTRACT： | OBJECTIVE To prepare Lonicerae japonicae flavonoids liposome membrane （LFLM）， and to study the mechanism of its anti-methicillin-resistant Staphylococcus aureus （MRSA）. METHODS L. japonicae flavonoids liposome （LFL） were prepared by ethanol injection with phospholipids and cholesterol as carriers. LFLM was prepared by loading LFL in mixed membranes of chitosan and polyvinyl alcohol. The particle size of LFL， polydispersity index （PDI） and release rate of LFLM were measured. A control group， LFLM low-concentration group （2.5 mg/mL）， LFLM-medium concentration group （5 mg/mL）， LFLM high-concentration group （10 mg/mL）， positive group （10 μg/mL vancomycin）， and combination therapy group （10 mg/mL LFLM and 10 μg/mL vancomycin） were set up. The effects of LFLM on MRSA colony formation and survival as well as the formation of MRSA biofilm were evaluated. The contents of K+， Mg2+， lactic dehydrogenase （LDH）， and alkaline phosphatase （AKP） in the supernatant of MRSA were determined. mRNA expressions of mecA and mecR1 in MRSA were determined. RESULTS The particle size of LFL was （80.91±3.96） nm， and the PDI was 0.26±0.07. The release rate of LFLM was 55% within 12 h and 73% within 36 h. Compared with the control group， the number of MRSA colony formation decreased significantly in LFLM medium-concentration and high-concentration groups， and the positive group （P＜0.05）. The ratio of dead to live bacteria， biofilm inhibition rate， and the contents of K+， Mg2+， LDH and AKP were increased significantly （P＜0.05）， while the mRNA expressions of mecA and mecR1 were decreased significantly （P＜0.05）. The combination of LFLM and vancomycin further enhanced the anti-MRSA effect. CONCLUSIONS LFLM is prepared successfully in the study and has good drug release characteristics. LFLM can exert anti-MRSA activity by promoting cell wall and membrane damage， and suppressing the mRNA expression of mecA and mecR1. |
| 期刊： | 2024年第35卷第21期 |
| 作者： | 熊瑞；张世鹏；刘恒旭；王璐；来小丹 |
| AUTHORS： | XIONG Rui，ZHANG Shipeng，LIU Hengxu，WANG Lu，LAI Xiaodan |
| 关键字： | 金银花黄酮；脂质体；膜剂；耐甲氧西林金黄色葡萄球菌；抗菌 |
| KEYWORDS： | Lonicerae japonicae flavonoids； liposomes； |
| 阅读数： | 3 次 |
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