Department of Ophthalmology, Dongguk University, Ilsan Hospital, Goyang, South Korea.
Transl Vis Sci Technol. 2023 Sep 1;12(9):23. doi: 10.1167/tvst.12.9.23.
To investigate the combined anti-Acanthamoeba effects of nitric oxide (NO) donors and hypochlorite to maximize amoebicidal outcomes while minimizing damage to human corneal epithelial cells (HCECs).
Acanthamoeba castellanii and primary cultured HCECs and keratocytes were treated with sodium hypochlorite (NaOCl), NO donors (sodium nitroprusside [SNP] and sodium nitrite [NaNO2]), or a combination of hypochlorite and NO donors. The viability of A. castellanii, HCECs, and keratocytes was assessed. Minimal inhibitory concentration (MIC) and fractional inhibitory concentration of NaOCl and NO donors were determined. The activation of mammalian targets of rapamycin (mTOR) and ERK and the expression of nitrite reductase and Nrf2 were assessed in HCECs using Western blot analysis. The cysticidal effects of combined NaOCl and NO donors were also evaluated.
A dose-dependent toxicity was observed in A. castellanii, HCECs, and keratocytes when treated with NaOCl and SNP. The range of tested NaNO2 concentrations showed no significant toxicity to HCECs; however, dose-dependent toxicity to A. castellanii was observed. The MIC of NaOCl against HCECs and A. castellanii was 8.0 mg/mL. The MIC of NaNO2 and SNP was 500 mM and 10 mM in both HCECs and A. castellanii, respectively. Weak attenuation of the mTOR and ERK phosphorylation was observed and Nrf2 expression decreased slightly after exposure of HCECs to 2.0 mg/mL NaOCl. For the combination treatment, NaOCl (0.125 mg/mL) was selected based on the safety of HCECs and the toxicity of A. castellanii. A more potent anti-Acanthamoeba effect and HCEC toxicity were observed when NaOCl was combined with SNP rather than NaNO2.
Combined NaOCl and NO donors had a stronger anti-Acanthamoeba effect compared to either drug alone.
This study demonstrates that the combined use of various drugs for the treatment of Acanthamoeba infection can enhance the anti-Acanthamoeba effect while minimizing the toxicity of the individual drug.
研究一氧化氮（NO）供体与次氯酸盐联合抗棘阿米巴的效果，在最大限度发挥杀阿米巴效果的同时，将对人角膜上皮细胞（HCEC）的损伤降到最低。
用次氯酸钠（NaOCl）、NO 供体（硝普钠[SNP]和亚硝酸钠[NaNO2]）或次氯酸盐和 NO 供体联合处理棘阿米巴原虫、原代培养的 HCEC 和角膜基质细胞。评估棘阿米巴原虫、HCEC 和角膜基质细胞的活力。测定 NaOCl 和 NO 供体的最小抑菌浓度（MIC）和部分抑菌浓度。用 Western blot 分析评估 HCEC 中哺乳动物雷帕霉素靶蛋白（mTOR）和细胞外信号调节激酶（ERK）的激活以及亚硝酸盐还原酶和 Nrf2 的表达。还评估了联合使用 NaOCl 和 NO 供体的杀囊效果。
用 NaOCl 和 SNP 处理时，棘阿米巴原虫、HCEC 和角膜基质细胞出现剂量依赖性毒性。测试的 NaNO2 浓度范围内对 HCEC 无明显毒性，但观察到棘阿米巴原虫的剂量依赖性毒性。NaOCl 对 HCEC 和棘阿米巴原虫的 MIC 为 8.0mg/mL。NaNO2 和 SNP 对 HCEC 和棘阿米巴原虫的 MIC 分别为 500mM 和 10mM。暴露于 2.0mg/mL NaOCl 后，HCEC 中 mTOR 和 ERK 磷酸化的减弱和 Nrf2 表达的轻微减少。基于 HCEC 的安全性和棘阿米巴原虫的毒性，选择 NaOCl（0.125mg/mL）进行组合治疗。当 NaOCl 与 SNP 联合使用时，而不是与 NaNO2 联合使用时，对棘阿米巴原虫的抗阿米巴效果和 HCEC 毒性更强。
与单独使用任何一种药物相比，联合使用 NaOCl 和 NO 供体具有更强的抗棘阿米巴效果。
本文是机器翻译，仅供参考。
