Department of Medical Zoology, Kyung Hee University School of Medicine, Seoul, Republic of Korea.
Biomedical Science Institute, Kyung Hee University, Seoul, Republic of Korea; and.
Cornea. 2020 Feb;39(2):245-249. doi: 10.1097/ICO.0000000000002201.
The aim of this study was to reduce the cytotoxicity and improve the amoebicidal effect of polyhexamethylene biguanide (PHMB) at low concentrations by combining it with histone deacetylase (HDAC) inhibitors.
To reduce the cytotoxic effect on human corneal epithelial (HCE) cells, the concentration of PHMB was reduced to 0.0002%. To enhance the amoebicidal effect of PHMB, HDAC inhibitors such as suberoylanilide hydroxamic acid, MS275, or MC1568 were combined with it. Acanthamoeba and HCE cells were treated with 3 combinations to evaluate the amoebicidal and cytotoxic effects. Microscopy and fluorescence-activated cell sorting analysis were performed to investigate the apoptotic cell death of Acanthamoeba by these combinatorial treatments.
The low concentration of PHMB (0.0002%) alone demonstrated no cytopathic effects (CPEs) on HCE cells. Three combinatorial treatments using 0.0002% PHMB with 10 μM suberoylanilide hydroxamic acid, 10 μM MS275, or 10 μM MC1568 showed higher amoebicidal effects on A. castellanii trophozoites than PHMB alone. Fluorescence-activated cell sorting analysis confirmed that HDAC inhibitors increased the apoptotic cell death of Acanthamoeba. Mild CPEs were observed from HCE cells cotreated with PHMB and the HDAC inhibitors after 24 hours of exposure.
Combinatorial treatments showed high amoebicidal effects on Acanthamoeba and low CPEs on HCE cells, which suggests their potential application for Acanthamoeba keratitis treatment.
本研究旨在通过与组蛋白去乙酰化酶（HDAC）抑制剂联合使用，降低聚六亚甲基双胍（PHMB）在低浓度下的细胞毒性，并提高其杀阿米巴效果。
为降低对人角膜上皮（HCE）细胞的细胞毒性作用，将 PHMB 浓度降低至 0.0002%。为增强 PHMB 的杀阿米巴效果，将 HDAC 抑制剂如 suberoylanilide hydroxamic acid、MS275 或 MC1568 与之联合使用。用 3 种组合处理棘阿米巴和 HCE 细胞，以评估其杀阿米巴和细胞毒性作用。通过显微镜和荧光激活细胞分选分析研究这些组合处理对棘阿米巴的细胞凋亡死亡的影响。
单独使用低浓度 PHMB（0.0002%）对 HCE 细胞无细胞病变效应（CPE）。用 0.0002% PHMB 与 10 μM suberoylanilide hydroxamic acid、10 μM MS275 或 10 μM MC1568 三种组合处理对 A. castellanii 滋养体的杀阿米巴效果均高于单独使用 PHMB。荧光激活细胞分选分析证实，HDAC 抑制剂增加了棘阿米巴的细胞凋亡死亡。在暴露 24 小时后，HCE 细胞与 PHMB 和 HDAC 抑制剂共同处理后观察到轻微的 CPE。
联合治疗对棘阿米巴具有高杀阿米巴效果，对 HCE 细胞的 CPE 较低，提示其在棘阿米巴角膜炎治疗方面具有潜在的应用前景。
