Chudzik Barbara, Czernel Grzegorz, Miaskowski Arkadiusz, Gagoś Mariusz
Department of Cell Biology, Institute of Biology and Biochemistry, Maria Curie-Skłodowska University, 20-033 Lublin, Poland.
Department of Biophysics, University of Life Sciences in Lublin, Akademicka 13, 20-950 Lublin, Poland.
Eur J Pharm Sci. 2017 Jan 15;97:9-21. doi: 10.1016/j.ejps.2016.10.040. Epub 2016 Nov 2.
The AmB-Cu(II) complex has recently been reported as an antifungal agent with reduced aggregation of AmB in aqueous solutions, increased anti C. albicans activity and lower toxicity against human cells in vitro. In the present work, investigations of the activity of the AmB-Cu (II) complex against fungal pathogens with varying susceptibility, including C. albicans and C. parapsilosis strains and intrinsically resistant A. niger, and cytotoxicity in normal human dermal fibroblasts (NHDF) in vitro were performed. For better understanding of the mechanism of reduced cytotoxicity and increased fungicidal activity, the influence of the AmB-Cu (II) complex on membrane integrity and accumulation of cellular reactive oxygen species (ROS) and mitochondrial superoxide was compared with that of conventional AmB. In the sensitive C. albicans and C. parapsilosis strains, the AmB-Cu(II) complex showed higher fungicidal activity (the MIC value was 0.35-0.7μg/ml for the AmB-Cu (II) complex, and 0.45-0.9μg/ml for Fungizone) due to increased induction of oxidative damage with rapid inhibition of the ability to reduce tetrazolium dye (MTT). In the NHDF cell line, the CC value was 30.13±1.53μg/ml for the AmB-Cu(II) complex and 17.46±1.24μg/ml for (Fungizone), therefore, the therapeutic index (CC/MIC) determined in vitro was 86.09-43.04 for the AmB-Cu(II) complex and 38.80-19.40 for Fungizone. The lower cytotoxicity of the AmB-Cu(II) complex in human cells resulted from lower accumulation of cellular and mitochondrial reactive oxygen species. This phenomenon was probably caused by the induction of successful antioxidant defense of the cells. The mechanism of the reduced cytotoxicity of the AmB-Cu(II) complex needs further investigation, but the preliminary results are very promising.
最近有报道称，两性霉素B-铜（II）复合物是一种抗真菌剂，它在水溶液中两性霉素B的聚集减少，对白色念珠菌的活性增强，且在体外对人类细胞的毒性较低。在本研究中，我们对两性霉素B-铜（II）复合物针对包括白色念珠菌和近平滑念珠菌菌株以及固有耐药的黑曲霉在内的不同敏感性真菌病原体的活性进行了研究，并检测了其对正常人皮肤成纤维细胞（NHDF）的体外细胞毒性。为了更好地理解细胞毒性降低和杀真菌活性增加的机制，将两性霉素B-铜（II）复合物与传统两性霉素B对膜完整性、细胞活性氧（ROS）积累和线粒体超氧化物的影响进行了比较。在敏感的白色念珠菌和近平滑念珠菌菌株中，两性霉素B-铜（II）复合物表现出更高的杀真菌活性（两性霉素B-铜（II）复合物的MIC值为0.35-0.7μg/ml，两性霉素B的MIC值为0.45-0.9μg/ml），这是由于氧化损伤诱导增加，同时四氮唑染料（MTT）还原能力受到快速抑制。在NHDF细胞系中，两性霉素B-铜（II）复合物的CC值为30.13±1.53μg/ml，两性霉素B的CC值为17.46±1.24μg/ml，因此，体外测定的治疗指数（CC/MIC）两性霉素B-铜（II）复合物为86.09-43.04，两性霉素B为38.80-19.40。两性霉素B-铜（II）复合物在人类细胞中较低的细胞毒性是由于细胞和线粒体活性氧的积累较少。这种现象可能是由细胞成功诱导抗氧化防御引起的。两性霉素B-铜（II）复合物细胞毒性降低的机制需要进一步研究，但初步结果非常有前景。
