Dihydroartemisinin (DHA)-loaded liposomes were prepared by ethanol injection and characterized. The results showed that the drug-loaded liposomes were spherical, with the average particle size of (429.0±12.1)nm, the encapsulation rate of (80.0±3.6)％, and the drug loading of (8.6±0.3)％. Then, the antibacterial effect of the product on drug-resistant Mycobacterium tuberculosis(DR-MTB) was tested by propidium iodide(PI)/SYTO9(a green fluorescent nucleic acid dye) double staining-flow cytometry, and the best inhibitory concentration was selected. The results showed that compared with DHA, drug-loaded liposomes had better bacteriostatic effect on DR-MTB at low concentration, and the highest bacteriostatic rate was (40.3±1.1)％ at 4 μg/ml. The minimum inhibitory concentrations of DHA and drugloaded liposomes were 32 and 0.5 μg/ml, respectively. Acid-fast staining, fluorescence staining, Gram staining and realtime fluorescence quantitative PCR were used to detect the effect of drug-loaded liposomes on DR-MTB morphology and rifampin(RIF)-resistance genes. The results of acid-fast staining and fluorescent staining showed that in comparison to the negative control group, drug-loaded liposomes group significantly reduced the number of DR-MTB, and in parallels with the disappearance of bacterial chain structure and the presentation of short rods. And Gram staining of DR-MTB in above group became easy. Real-time fluorescence quantitative PCR results showed that the RIF-resistance genes of DRMTB were not reversed by drug-loaded liposomes, suggesting that its antibacterial mechanism might be related to the destruction of DR-MTB cell wall structure.
Key words
dihydroartemisinin /
nanomedicine /
drug-resistant Mycobacterium tuberculosis /
drug resistance gene /
inhibition rate
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] 余卫业, 谭卫国, 罗一婷, 等．2018 WHO全球结核报告: 全球与中国关键数据分析[J]．新发传染病电子杂志, 2018,3(4): 228-233．
[2] MAITRE T, AUBRY A, JARLIER V, et al．Multidrug and extensively drug-resistant tuberculosis [J]．Med Mal Infect,2017, 47(1): 3-10．
[3] 祝平凡．双氢青蒿素的体外稳定性及其时间依赖性药动学研究[D]．济南: 山东大学硕士学位论文, 2014．
[4] WANT M Y, ISLAMUDDIN M, CHOUHAN G, et al．Therapeutic efficacy of artemisinin-loaded nanoparticles in experimental visceral leishmaniasis [J]．Clloids Surf B Biointerfaces, 2015, 130: 215-221．
[5] UPADHYAY S, KHAN I, GOTHWAL A, et al．Conjugated and entrapped HPMA-PLA nano-polymeric micelles based dual delivery of first line anti TB drugs: Improved and safe drug delivery against sensitive and resistant Mycobacterium tuberculosis [J]．Pharm Res, 2017, 34(9): 1944-1955．
[6] 金丽霞．纳米药物载体的研究及临床应用[J]．中国组织工程研究与临床康复, 2010, 14(8): 1429-1432．
[7] 蔡丽丽．纳米壳聚糖双氢青蒿素对感染耐药结核杆菌巨噬细胞磷脂酰肌醇信号通路的影响[D]．泸洲: 西南医科大学硕士学位论文, 2017．
[8] MOHAMED M, ABU LILA A S, SHIMIZU T, et al．PEGylated liposomes: immunological responses [J]．Sci Technol Adv Mater, 2019, 20(1): 710-724．
[9] 黄玉平, 屈亚虹, 马振亚, 等．应用线性探针技术快速诊断耐多药肺结核效果分析[J]．中国防痨杂志, 2011, 33(11):722-724．
[10] KALANI K, ALAM S, CHATURVEDI V, et al．In vitro, in silico and ex vivo studies of dihydroartemisinin derivatives
as antitubercular agents [J]．Curr Top Med Chem, 2019,19(8): 633-644．
[11] VAN ZYL L, VILJOEN J M, HAYNES R K, et al．Topical delivery of artemisone, clofazimine and decoquinate encapsulated in vesicles and their in vitro efficacy against Mycobacterium tuberculosis [J]．AAPS PharmSciTech,2019, 20(1): 33．
[12] 金 一, 李 博, 李 贞, 等．一种双氢青蒿素磷脂复合物及其制备与应用: 中国, 102078317A [P]．2011-06-01．
[13] 赵凯存, 宋振玉．双氢青蒿素在人的药代动力学及与青蒿素的比较[J]．药学学报, 1993, 28(5): 342-346．
[14] 董 娜, 付玉荣, 伊正君．结核分枝杆菌感染致脂代谢异常及其机制研究进展[J]．中华结核和呼吸杂志, 2016,39(7): 548-550．
[15] LIN L, MAO X, SUN Y, et al．Antibacterial mechanism of artemisinin/beta-cyclodextrins against methicillin-resistant
Staphylococcus aureus (MRSA)[J]．Microb Pathog, 2018,118: 66-73．
[16] 汤自豪, 周小鸥, 高兴政, 等．双氢青蒿素、甲硝唑及两药联合应用体外抗阴道毛滴虫效果的研究[J]．中国医科大学学报, 2008, 37(2): 200-203．
[17] MESHNICK S R．Artemisinin: mechanisms of action,resistance and toxicity [J]．Int J Parasitol, 2002, 32(13):1655-1660．
[18] MESHNICK S R, YANG Y Z, LIMA V, et al．Irondependent free radical generation from the antimalarial agent artemisinin (qinghaosu) [J]．Antimicrob Agents Chemother, 1993, 37(5): 1108-1114．
[19] 詹能勇, 聂 广, 黄慧谦, 等．青蒿琥酯及二氢青蒿素逆转耐药结核分枝杆菌的初步研究[J]．中山大学学报: 医学科学版, 2009, 30(3S): 8-11．
{{custom_fnGroup.title_en}}
Footnotes
{{custom_fn.content}}
PDF(2647 KB)
