体外和体内研究杨梅素与硝酸咪康唑联合应用于温敏水凝胶对白色念珠菌生物膜的作用。

In vitro and in vivo effects of the combination of myricetin and miconazole nitrate incorporated to thermosensitive hydrogels, on C. albicans biofilms.

机构信息

School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an, 710061, P. R. China.

Biobank, The first affiliated hospital of Xi'an Jiaotong University, Xi'an, 710054, P. R. China.

出版信息

Phytomedicine. 2020 Jun;71:153223. doi: 10.1016/j.phymed.2020.153223. Epub 2020 May 12.

DOI:10.1016/j.phymed.2020.153223

PMID:32460204

Abstract

BACKGROUND

Candida albicans-related infections are common infections in clinic, among which biofilm-associated infections are most devastating and challenging to overcome. Myricetin (MY) is a plant-derived natural product with various pharmacological activities. Its anti-biofilm effect against C. albicans and its ability to increase the antifungal effect of miconazole nitrate (MN) were unclear and yet need to be explored.

HYPOTHESIS/PURPOSE: In this study the anti-biofilm effect of MY and its ability to increase the antifungal effect of MN were investigated in vitro and in vivo.

STUDY DESIGN AND METHODS

MY or/and MN were incorporated into a thermosensitive hydrogel (TSH) of poloxamer. The safety of MY or/and MN loaded TSHs towards human umbilical vein endothelial cells (HUVEC) was evaluated by a MTT assay and the in vivo safety towards mice knees was confirmed by histopathological examination. The anti-biofilm effect of MY and its ability to increase the antifungal effect of MN were investigated in vitro with C. albicans ATCC 10231 by broth microdilution method, crystal violet staining and scanning electron microscopy (SEM), as well as in vivo in an established mouse model of periprosthetic joint infection (PJI) by SEM, histological analysis, microorganism culture and detection of the serum levels of interleukin-6 (IL-6). The mechanism of action of MY was analyzed by qRT-PCR assay with C. albicans SC5314.

RESULTS

Our results showed that MY and MN incorporated into TSHs exhibited good stability and safety, excellent temperature sensitivity and controlled drug release property. MY (5-640 µg/ml) exhibited no effect on C. albicans cell viability and MY (≥80 µg/ml) showed a significantly inhibitory effect on biofilm formation. MIC (the lowest concentrations of drugs resulting in 50% decrease of C. albicans growth) and MIC (the lowest concentrations of drugs resulting in 80% decrease of C. albicans growth) of MN were respectively decreased from 2 µg/ml to 0.5 µg/ml and from 4 µg/ml to 2 µg/ml when used in combination with MY (80 µg/ml). The mouse PJI was effectively prevented by MY and MN incorporated into TSH.

CONCLUSIONS

Local application of MY and MN incorporated into TSH might be useful for clinical biofilm-associated infections.

摘要

背景

白色念珠菌相关感染是临床常见感染，其中生物膜相关感染最具破坏性且难以克服。杨梅素（MY）是一种具有多种药理活性的植物源性天然产物。其抗白色念珠菌生物膜作用及其增强硝酸咪康唑（MN）抗真菌作用尚不清楚，有待进一步研究。

假设/目的：本研究旨在体外和体内研究 MY 的抗生物膜作用及其增强 MN 抗真菌作用的能力。

研究设计和方法

将 MY 或/和 MN 掺入泊洛沙姆的温敏水凝胶（TSH）中。通过 MTT 测定法评估 MY 或/和 MN 负载 TSH 对人脐静脉内皮细胞（HUVEC）的安全性，并通过组织病理学检查确认其对小鼠膝关节的体内安全性。通过肉汤微量稀释法、结晶紫染色和扫描电子显微镜（SEM）研究 MY 和 MN 对白色念珠菌 ATCC 10231 的抗生物膜作用及其增强 MN 抗真菌作用的能力，以及通过 SEM、组织学分析、微生物培养和检测白细胞介素-6（IL-6）的血清水平，在建立的假体周围关节感染（PJI）小鼠模型中进行体内研究。通过 qRT-PCR 分析白色念珠菌 SC5314 分析 MY 的作用机制。

结果

结果表明，掺入 TSH 的 MY 和 MN 表现出良好的稳定性和安全性、优异的温度敏感性和控制药物释放性能。MY（5-640 µg/ml）对白色念珠菌细胞活力无影响，MY（≥80 µg/ml）对生物膜形成有显著抑制作用。MN 的最低药物浓度（导致白色念珠菌生长减少 50%的药物浓度）和 MN 的最低药物浓度（导致白色念珠菌生长减少 80%的药物浓度）分别从 2 µg/ml 降低至 0.5 µg/ml 和从 4 µg/ml 降低至 2 µg/ml 当与 MY（80 µg/ml）联合使用时。掺入 TSH 的 MY 和 MN 可有效预防小鼠 PJI。