Zlotnikov Igor D, Ezhov Alexander A, Vigovskiy Maksim A, Grigorieva Olga A, Dyachkova Uliana D, Belogurova Natalia G, Kudryashova Elena V
Faculty of Chemistry, Lomonosov Moscow State University, Leninskie Gory, 1/3, 119991 Moscow, Russia.
Faculty of Physics, Lomonosov Moscow State University, Leninskie Gory, 1/2, 119991 Moscow, Russia.
Diagnostics (Basel). 2023 Feb 12;13(4):698. doi: 10.3390/diagnostics13040698.
Visualization of the interaction of drugs with biological cells creates new approaches to improving the bioavailability, selectivity, and effectiveness of drugs. The use of CLSM and FTIR spectroscopy to study the interactions of antibacterial drugs with latent bacterial cells localized in macrophages create prospects to solve the problems of multidrug resistance (MDR) and severe cases. Here, the mechanism of rifampicin penetration into bacterial cells was studied by tracking the changes in the characteristic peaks of cell wall components and intracellular proteins. However, the effectiveness of the drug is determined not only by penetration, but also by efflux of the drugs molecules from the bacterial cells. Here, the efflux effect was studied and visualized using FTIR spectroscopy, as well as CLSM imaging. We have shown that because of efflux inhibition, eugenol acting as an adjuvant for rifampicin showed a significant (more than three times) increase in the antibiotic penetration and the maintenance of its intracellular concentration in (up to 72 h in a concentration of more than 2 μg/mL). In addition, optical methods have been applied to study the systems containing bacteria localized inside of macrophages (model of the latent form), where the availability of bacteria for antibiotics is reduced. Polyethylenimine grafted with cyclodextrin carrying trimannoside vector molecules was developed as a drug delivery system for macrophages. Such ligands were absorbed by CD206+ macrophages by 60-70% versus 10-15% for ligands with a non-specific galactose label. Owing to presence of ligands with trimannoside vectors, the increase in antibiotic concentration inside macrophages, and thus, its accumulation into dormant bacteria, is observed. In the future, the developed FTIR+CLSM techniques would be applicable for the diagnosis of bacterial infections and the adjustment of therapy strategies.
可视化药物与生物细胞的相互作用为提高药物的生物利用度、选择性和有效性创造了新方法。利用共聚焦激光扫描显微镜（CLSM）和傅里叶变换红外光谱（FTIR）研究抗菌药物与巨噬细胞内潜伏细菌细胞的相互作用，为解决多重耐药性（MDR）问题和重症病例带来了希望。在此，通过追踪细胞壁成分和细胞内蛋白质特征峰的变化，研究了利福平进入细菌细胞的机制。然而，药物的有效性不仅取决于其渗透，还取决于药物分子从细菌细胞中的流出。在此，利用FTIR光谱以及CLSM成像研究并可视化了流出效应。我们已经表明，由于流出抑制，作为利福平佐剂的丁香酚使抗生素的渗透显著增加（超过三倍），并且其细胞内浓度得以维持（在浓度超过2μg/mL时可达72小时）。此外，光学方法已被应用于研究含有巨噬细胞内细菌（潜伏形式模型）的系统，在该系统中细菌对抗生素的可及性降低。接枝有携带三甘露糖苷载体分子的环糊精的聚乙烯亚胺被开发为巨噬细胞的药物递送系统。与带有非特异性半乳糖标签的配体相比，此类配体被CD206+巨噬细胞吸收的比例为60 - 70%，而后者为10 - 15%。由于存在带有三甘露糖苷载体的配体，观察到巨噬细胞内抗生素浓度增加，进而其在休眠细菌中的积累也增加。未来，所开发的FTIR + CLSM技术将适用于细菌感染的诊断和治疗策略的调整。
