荆皮癣湿酊诱导红色毛癣菌凋亡的机制研究

Jingpixian tincture induces apoptosis of Trichophyton rubrum

摘要:

【目的】本研究旨在探讨复方中药荆皮癣湿酊(Jingpixian tincture, JPXT)对红色毛癣菌(Trichophyton rubrum)的凋亡诱导作用，以阐明其可能的抗真菌作用机制。【方法】采用细胞计数试剂盒-8 (cell counting kit-8, CCK-8)评价荆皮癣湿酊对红色毛癣菌生长活力的影响；流式细胞仪检测红色毛癣菌细胞内活性氧(reactive oxygen species, ROS)水平和线粒体膜电位(mitochondrial membrane potential, MMP)变化；Annexin V-FITC/PI染色荧光显微镜观察红色毛癣菌细胞磷脂酰丝氨酸(phosphatidylserine, PS)外翻情况；流式细胞术检测红色毛癣菌细胞凋亡率；FITC-VAD-FMK染色观察红色毛癣菌偏半胱天冬酶(metacaspase)活性；紫外分光光度计测定红色毛癣菌细胞色素C氧化酶的活性。【结果】荆皮癣湿酊处理后的红色毛癣菌细胞活力与MMP水平均有所降低，ROS水平显著升高，PS外翻与凋亡率明显增加，偏半胱天冬酶活性显著升高，细胞色素C氧化酶活性降低。【结论】荆皮癣湿酊可通过诱导菌体凋亡的方式发挥对红色毛癣菌的抗菌作用。

Abstract:

[Objective] To investigate the role of Jingpixian tincture (JPXT) in inducing apoptosis of Trichophyton rubrum and reveal the antifungal mechanism of JPXT. [Methods] Cell counting kit-8 (CCK-8) was used to evaluate the effect of JPXT on the viability of T. rubrum. The reactive oxygen species (ROS) level and mitochondrial membrane potential (MMP) in T. rubrum were measured by flow cytometry. Annexin V-FITC/PI staining was employed to observe the externalization of phosphatidylserine (PS) in T. rubrum. Flow cytometry, FITC-VAD-FMK staining, and an ultraviolet spectrophotometer were employed to determine the apoptosis rate, metacaspase activity, and cytochrome C oxidase activity, respectively. [Results] After JPXT treatment, the viability and MMP of T. rubrum decreased, and the ROS level increased. Furthermore, the JPXT treatment promoted PS externalization, increased the apoptosis rate and metacaspase activity, and decreased the cytochrome C oxidase activity. [Conclusion] JPXT can inhibit T. rubrum by inducing apoptosis.