Page 46 - 《中国药房》2024年21期

金银花黄酮脂质体膜的制备及其抗MRSA的机制研究 Δ

熊 瑞 ，张世鹏，刘恒旭，王 璐，来小丹 [陆军军医大学第一附属医院江北院区（陆军第九五八医院）药剂科，
#
＊
重庆 400020]

中图分类号 R943；R965 文献标志码 A 文章编号 1001-0408（2024）21-2616-06
DOI 10.6039/j.issn.1001-0408.2024.21.07

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（2.5 mg/mL）、LFLM中浓度组（5
mg/mL）、LFLM 高浓度组（10 mg/mL）、阳性组（10 μg/mL 万古霉素）和联合用药组（10 mg/mL LFLM+10 μg/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 、Mg 、乳酸脱氢酶（LDH）、碱性磷酸酶（AKP）的含
+
2+
量；检测 MRSA 中 mecA、mecR1 mRNA 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12 h内的释放率为55%，36 h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 、Mg 、LDH、AKP 含量均显著升高（P＜0.05），mecA、mecR1
2+
+
mRNA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、mecR1 mRNA表达，从而发挥抗菌作用。
关键词 金银花黄酮；脂质体；膜剂；耐甲氧西林金黄色葡萄球菌；抗菌

Preparation of Lonicerae japonicae flavonoids liposome membrane and its antibacterial mechanism against
methicillin-resistant Staphylococcus aureus
XIONG Rui，ZHANG Shipeng，LIU Hengxu，WANG Lu，LAI Xiaodan[Dept. of Pharmacy， Jiangbei Campus of
the First Affiliated Hospital of Army Medical University （The 958th Hospital of PLA）， Chongqing 400020，
China]

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 ， Mg ， lactic dehydrogenase （LDH）， and alkaline phosphatase
2+
+
（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 ， Mg ， LDH and AKP were increased significantly （P＜0.05）， while the
+
2+
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
Δ 基金项目 重庆市科卫联合医学科研项目（No.2022MSXM187） promoting cell wall and membrane damage， and suppressing
＊第一作者 主管药师，硕士。研究方向：中药药理学及循证药学。
the mRNA expression of mecA and mecR1.
E-mail：rxiong2017@sina.com
# 通信作者 副主任药师，硕士。研究方向：药剂学及循证药学。 KEYWORDS Lonicerae japonicae flavonoids； liposomes；
E-mail：laixiaodan0926@sina.com membrane； Staphylococcus aureus； antibacterial

· 2616 · China Pharmacy 2024 Vol. 35 No. 21 中国药房 2024年第35卷第21期