金银花总黄酮脂质体的制备及对MRSA的抑制作用

熊瑞; 倪睿; 刘恒旭; 张世鹏; 王璐; 来小丹

doi:10.16016/j.2097-0927.202405014

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金银花总黄酮脂质体的制备及对MRSA的抑制作用

药学 | 更新时间：2026-01-09

- 金银花总黄酮脂质体的制备及对MRSA的抑制作用
- Preparation of total flavonoid-liposomes of Lonicerae Japonicae flos and their inhibitory effect on methicillin-resistant Staphylococcus aureus
- 陆军军医大学学报 2024年46卷第24期页码：2755-2764
- 作者机构：
  
  1.400020 重庆，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
  2.400042 重庆，陆军特色医学中心(大坪医院)药剂科
- 作者简介：
  
  [来小丹，E-mail：laixiaodan0926@sina.comLAI Xiaodan, E-mail: laixiaodan0926@sina.com
- 基金信息：
  
  重庆市科卫联合中医药项目(2022MSXM187)
- DOI：10.16016/j.2097-0927.202405014
  中图分类号： R285.5;R378.11;R944.9
- 收稿：2024-05-06，
  
  修回：2024-07-20，
  
  纸质出版：2024-12-30
- 稿件说明：
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熊瑞, 倪睿, 刘恒旭, 等. 金银花总黄酮脂质体的制备及对MRSA的抑制作用[J]. 陆军军医大学学报, 2024,46(24):2755-2764.

Rui XIONG, Rui NI, Hengxu LIU, et al. Preparation of total flavonoid-liposomes of Lonicerae Japonicae flos and their inhibitory effect on methicillin-resistant Staphylococcus aureus[J]. Journal of Army Medical University, 2024, 46(24): 2755-2764.
熊瑞, 倪睿, 刘恒旭, 等. 金银花总黄酮脂质体的制备及对MRSA的抑制作用[J]. 陆军军医大学学报, 2024,46(24):2755-2764. DOI： 10.16016/j.2097-0927.202405014.

Rui XIONG, Rui NI, Hengxu LIU, et al. Preparation of total flavonoid-liposomes of Lonicerae Japonicae flos and their inhibitory effect on methicillin-resistant Staphylococcus aureus[J]. Journal of Army Medical University, 2024, 46(24): 2755-2764. DOI： 10.16016/j.2097-0927.202405014.

摘要

目的

制备金银花总黄酮(

Lonicerae Japonicae

Flos total flavonoids

LJFTF)脂质体

并评价其对耐甲氧西林金黄色葡萄球菌(methicillin-resistant

Staphylococcus aureus

MRSA)的抑制作用。

方法

采用乙醇注入法制备LJFTF脂质体(LJFTF liposome

LJFTFL)

以磷脂浓度、磷脂与胆固醇质量比(脂胆比)、LJFTF质量浓度为考察因素

以包封率、载药量为评价指标

通过星点设计-效应面法优化制备工艺。通过观察性状、利用激光粒度仪测定粒径、多分散指数(Polymer dispersity index

PDI)和Zeta电位对LJFTFL进行表征

并通过高效液相色谱法测定其体外释放率。采用平板法和细菌活死染色实验观察LJFTFL对MRSA的抗菌活性

采用结晶紫染色观察LJFTFL对MRSA生物膜的抑制作用。

结果

经过优化和验证确定LJFTFL的最佳处方为: 磷脂浓度35.00 mg/mL

脂胆比15.00

LJFTF浓度5.74 mg/mL; 制得的LJFTFL呈淡黄色乳状液

测得其平均包封率为86.37%(RSD=0.37%

=3)

平均载药量为11.69%(RSD=0.09%

=3)

粒径为(173.60±1.07) nm

PDI值为0.15±0.05

Zeta电位为(-4.86±0.60) mV; 游离的LJFTF在4 h内释放完全

LJFTFL在体外12 h的累计释放率为(59.44±3.58)%

在30 h的累计释放量为(63.58±5.78)%。平板法和细菌活死染色实验表明

LJFTFL可显著抑制MRSA生长并促进其死亡; 结晶紫染色结果显示

LJFTFL可显著抑制MRSA生物膜形成

且LJFTFL干预组与同浓度游离的LJFTF干预组相比差异具有统计学意义(

0.001)。

结论

本研究成功制得LJFTFL并对其处方工艺进行了优化

LJFTFL在体外对MRSA呈显著的抗菌增效活性。

Abstract

Objective

To prepare the liposomes of total flavonoids from

Lonicerae Japonicae

flos (LJFTF) and evaluate their inhibitory effect on methicillin-resistant

Staphylococcus aureus

(MRSA).

Methods

LJFTF liposomes were prepared by ethanol injection.Then the preparation process was optimized by star point design-effect surface method

with the concentrations of phospholipid

mass ratio of phospholipid to cholesterol and mass concentration of LJFTF as the influencing factors

and the encapsulation rate and drug-loading rate as evaluation indicators.The obtained liposomes were characterized by observing their properties

measuring particle size

polymer dispersity index (PDI) and Zeta potential through laser particle size analyzer.High performance liquid chromatography (HPLC) was used to determine the release rate of the liposomes.The antibacterial activity of the liposomes against MRSA was observed by plate method and live/dead staining

and the inhibitory effect of the liposomes on MRSA biofilm was observed by crystal violet staining.

Results

After optimization and verification

the best preparation process was as follows: phospholipid concentration 35.0 mg/mL

phospholipid to cholesterol mass ratio 15.0

and LJFTF concentration 5.7 mg/mL.The prepared liposomes were light yellow emulsion

with an average encapsulation rate of 86.37%(RSD=0.37%

=3)

and an average drug-loading rate of 11.69%(RSD=0.09%

=3)

a particle size of 173.60±1.07 nm

a PDI value of 0.15±0.05

and a Zeta potential of-4.86±0.60 mV.The free LJFTF was completely released within 4 h from the dialysis bag

and the

in vitro

cumulative release of LJFTF was (59.44±3.58)% at 12 h and (63.58±5.78)% at 30 h.Plate and live/dead staining showed that the prepared liposomes significantly inhibited the growth and promoted the death of MRSA strain.Crystal violet staining displayed that the liposomes significantly inhibited the formation of MRSA biofilm (

0.001)

with significant difference in comparison with same dose of LJFTF (

0.001).

Conclusion

LJFTF liposomes are successfully prepared

and the formulation process has been optimized.The liposomes show significant anti-MRSA activity

in vitro

关键词

Keywords

references

WANG B J, XU Y L, ZHAO H L, et al . Methicillin-resistant Staphylococcus aureus in China: a multicentre longitudinal study and whole-genome sequencing[J ] . Emerg Microbes Infect, 2022, 11(1): 532-542. DOI: 10.1080/22221751.2022.2032373..

刘婧怡, 蔡仑, 段浩, 等. 大黄素抑制MRSA生物膜活性及其作用机制研究[J]. 陆军军医大学学报, 2022, 44(7): 684-690. DOI: 10.16016/j.2097-0927.202112068.

LIU J Y, CAI L, DUAN H, et al . Inhibitory effect of emodin on MRSA biofilm and its mechanism[J ] . J Army Med Univ, 2022, 44(7): 684-690. DOI: 10.16016/j.2097-0927.202112068.

HSIEH R C, LIU R, BURGIN D J, et al . Understanding mechanisms of virulence in MRSA: implications for antivirulence treatment strategies[J ] . Expert Rev Anti Infect Ther, 2023, 21(9): 911-928. DOI: 10.1080/14787210.2023.2242585.

TONG S Y C, LYE D C, YAHAV D, et al . Effect of vancomycin or daptomycin with vs without an antistaphylococcal β-lactam on mortality, bacteremia, relapse, or treatment failure in patients with MRSA bacteremia: a randomized clinical trial[J ] . JAMA, 2020, 323(6): 527-537. DOI: 10.1001 /jama.2020.0103.

吴娇, 王聪, 于海川. 金银花中的化学成分及其药理作用研究进展[J]. 中国实验方剂学杂志, 2019, 25(4): 225-234. DOI: 10.13422/j.cnki.syfjx.20190408.

WU J, WANG C, YU H C. Chemical constituents and pharmacological effect of Lonicerae japonicae Flos [J ] . Chin J Exp Tradit Med Formulae, 2019, 25(4): 225-234. DOI: 10.13422/j.cnki.syfjx.20190408.

SHANG X F, PAN H, LI M X, et al . Lonicera japonica Thunb.: ethnopharmacology, phytochemistry and pharmacology of an important traditional Chinese medicine[J ] . J Ethnopharmacol, 2011, 138(1): 1-21. DOI: 10.1016/j.jep.2011.08.016..

罗旋, 王薇, 胡本祥, 等. 金银花抗病毒化学成分及作用研究进展[J]. 陕西中医药大学学报, 2024, 47(2): 138-143. DOI: 10.13424/j.cnki.jsctcm.2024.02.027.

LUO X, WANG W, HU B X, et al . Research progress on antiviral chemical constituents and functions of honeysuckle[J ] . J Shaanxi Univ Chin Med, 2024, 47(2): 138-143. DOI: 10.13424/j.cnki.jsctcm.2024.02.027.

LI Y K, LI W, FU C M, et al . Lonicerae japonicae Flos and Lonicerae flos : a systematic review of ethnopharmacology, phytochemistry and pharmacology[J ] . Phytochem Rev, 2020, 19(1): 1-61. DOI: 10.1007/s11101-019-09655-7..

MENG Y N, NIU X, LI G L. Liposome nanoparticles as a novel drug delivery system for therapeutic and diagnostic applications[J]. Curr Drug Deliv, 2022, 20(1): 41-56. DOI: 10.2174/1567201819666220324093821.

AJEESHKUMAR K K, ANEESH P A, RAJUN, et al . Advancements in liposome technology: preparation techniques and applications in food, functional foods, and bioactive delivery: a review[J ] . Compr Rev Food Sci Food Saf, 2021, 20(2): 1280-1306. DOI: 10.1111/1541-4337.12725.

BOAFO G F, MAGAR K T, EKPO M D, et al . The role of cryoprotective agents in liposome stabilization and preservation[J ] . Int J Mol Sci, 2022, 23(20): 12487. DOI: 10.3390/ijms232012487.

王兴芝, 代英辉, 王东凯. 脂质体的制备方法及应用的研究进展[J]. 中国药剂学杂志(网络版), 2024, 22(1): 14-24. DOI: 10.14146/j.cnki.cjp.2024.01.003.

WANG X Z, DAI Y H, WANG D K. Research progress in preparation methods and applications of liposomes[J]. Chin J Pharm Online Ed, 2024, 22(1): 14-24. DOI: 10.14146/j.cnki.cjp.2024.01.003.

SHEORAN R, KHOKRA S L, CHAWLA V, et al . Recent patents, formulation tech niques, classification and characterization of liposomes[J ] . Recent Pat Nanotechnol, 2019, 13(1): 17-27. DOI: 10.2174/1872210513666 181127110413.

马晓星, 隋小宇, 刘畅, 等. Box-Behnken响应面法优化厚朴酚与小檗碱共载脂质体的处方工艺[J]. 中国医院药学杂志, 2024, 44(7): 796-801, 858. DOI: 10.13286/j.1001-5213.2024.07.08.

MA X X, SUI X Y, LIU C, et al . Formulation optimization of berberine and magnolol co-loaded liposomes by Box-Behnken response surface method[J ] . Chin J Hosp Pharm, 2024, 44(7): 796-801, 858. DOI: 10.13286/j.1001-5213.2024.07.08.

苏艳莹, 雷小小, 桂卉, 等. 星点设计-效应面法优化山茱萸总环烯醚萜苷脂质体的制备工艺[J]. 中国药房, 2018, 29(19): 2612-2616. DOI: 10.6039/j.issn.1001-0408.2018.19.05.

SU Y Y, LEI X X, GUI H, et al . Optimization of preparation technology of total iridoid glycosides liposome of Cornus officinalis by central composite design-response surface methodology[J ] . China Pharm, 2018, 29(19): 2612-2616. DOI: 10.6039/j.issn.1001-0408.2018.19.05..

邰克东, 赵苏茂, 杨紫恒, 等. 高压均质对脂质体囊泡特性和稳定性的影响[J]. 食品科学, 2019, 40(17): 169-177. DOI: 10.7506/spkx1002-6630-20180916-158.

TAI K D, ZHAO S M, YANG Z H, et al . Effect of high-pressure homogenization on vesicle characteristics and stability of liposomes[J ] . Food Sci, 2019, 40(17): 169-177. DOI: 10.7506/spkx1002-6630-20180916-158.

LE K Y, OTTO M. Approaches to combating methicillin-resistant Staphylococcus aureus (MRSA) biofilm infections[J ] . Expert Opin Investig Drugs, 2024, 33(1): 1-3. DOI: 10.1080/13543784.2024.2305136.

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