Abstract

There are an estimated 1 billion cases of superficial fungal infection globally. Fungal pathogens form biofilms within wounds and delay the wound healing process. Miconazole and terbinafine are commonly used to treat fungal infections. They induce the accumulation of reactive oxygen species (ROS) in fungi, resulting in the death of fungal cells. ROS are highly reactive molecules, such as oxygen (O₂), superoxide anion (O₂•−), hydrogen peroxide (H₂O₂) and hydroxyl radicals (•OH). Although ROS generation is useful for killing pathogenic fungi, it is cytotoxic to human keratinocytes. To the best of our knowledge, the effect of miconazole and terbinafine on HaCaT cells has not been studied with respect to intracellular ROS stimulation. We hypothesized that miconazole and terbinafine have anti-wound healing effects on skin cells when used in antifungal treatment because they generate ROS in fungal cells. We used sulforhodamine B protein staining to investigate cytotoxicity and 2′,7′-dichlorofluorescein diacetate to determine ROS accumulation at the 50% inhibitory concentrations of miconazole and terbinafine in HaCaT cells. Our preliminary results showed that topical treatment with miconazole and terbinafine induced cytotoxic responses, with miconazole showing higher cytotoxicity than terbinafine. Both the treatments stimulated ROS in keratinocytes, which may induce oxidative stress and cell death. This suggests a negative correlation between intracellular ROS accumulation in keratinocytes treated with miconazole or terbinafine and the healing of fungi-infected skin wounds.

摘要

全球估计有 10 亿例浅表真菌感染病例。真菌病原体在伤口内形成生物膜并延迟伤口愈合过程。咪康唑和特比萘芬常用于治疗真菌感染。它们诱导真菌中活性氧（ROS）的积累，导致真菌细胞死亡。ROS 是高活性分子，如氧 (O ₂ )、超氧阴离子 (O ₂ •−)、过氧化氢 (H ₂ O ₂) 和羟基自由基 (•OH)。尽管 ROS 生成可用于杀死病原真菌，但它对人类角质形成细胞具有细胞毒性。据我们所知，尚未就细胞内 ROS 刺激研究咪康唑和特比萘芬对 HaCaT 细胞的影响。我们假设咪康唑和特比萘芬在用于抗真菌治疗时对皮肤细胞具有抗伤口愈合作用，因为它们在真菌细胞中产生活性氧。我们使用磺基罗丹明 B 蛋白染色来研究细胞毒性，并使用 2',7'-二氯荧光素二乙酸酯来确定 HaCaT 细胞中 50% 抑制浓度的咪康唑和特比萘芬时的 ROS 积累。我们的初步结果表明，用咪康唑和特比萘芬局部治疗会引起细胞毒性反应，咪康唑比特比萘芬表现出更高的细胞毒性。两种治疗都刺激了角质形成细胞中的 ROS，这可能会诱导氧化应激和细胞死亡。这表明用咪康唑或特比萘芬处理的角质形成细胞中细胞内 ROS 积累与真菌感染皮肤伤口的愈合之间存在负相关。