Shikonin treats Aspergillus fumigatus keratitis by activating autophagy and reducing fungal load.

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research Pub Date : 2025-10-01 Epub Date: 2025-08-06 DOI:10.1016/j.exer.2025.110569

Xuguang Yang, Fan Cong, Lingwen Gu, Qian Wang, Lina Zhang, Jing Lin, Weichen Liu, Xiaofeng Yao, Cui Li, Guiqiu Zhao

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引用次数: 0

Abstract

The objective of this study was to explore the therapeutic potential of Shikonin (SK) in the treatment of fungal keratitis. Various tests conducted in this study were fluorescein sodium staining, Cell Counting Kit-8 (CCK-8), RT-PCR, Western blot (WB), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) methods. And the autophagic and phagocytic functions of RAW 264.7 cells were examined using transmission electron microscopy (TEM) and phagocytosis assay. The efficacy of SK in inhibiting Aspergillus fumigatus (A. fumigatus) was confirmed by determining the minimal inhibitory concentration (MIC), crystal violet staining assay, calcofluor white staining (CFW), propidium iodide absorption assay (PI), and fungal adhesion assay. Compared to the group treated with 0.1 % DMSO, the group treated with SK demonstrated a significant decrease in clinical score and the levels of IL-6, TNF-α, and IL-1β. SK also promoted the activity of autophagy proteins LC3B- II/I and Beclin-1 while reducing P62 levels. After treating with SK, there was an increase in autolysosomes in RAW 264.7 cells, accompanied by enhanced phagocytosis of fungi and conidia. Pretreatment with 3-methyladenine (3-MA) counteracted the ability of SK to reduce inflammatory markers. Furthermore, SK inhibited A. fumigatus growth, damaged cell membrane, impaired biofilms, and suppressed conidial adhesion. In conclusion, SK exhibits anti-inflammatory function by activating autophagy and anti-fungal function by reducing fungal load, highlighting its potential treatment effect for fungal keratitis.

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紫草素通过激活自噬和减少真菌负荷治疗烟曲霉角膜炎。

本研究的目的是探讨紫草素（SK）治疗真菌性角膜炎的治疗潜力。本研究采用荧光素钠染色、细胞计数试剂盒-8 （CCK-8）、RT-PCR、western blot （WB）、免疫荧光和酶联免疫吸附试验（ELISA）等方法。采用透射电镜（TEM）和吞噬实验检测RAW 264.7细胞的自噬和吞噬功能。通过最小抑菌浓度（MIC）测定、结晶紫染色法、荧光白染色法（CFW）、碘化丙啶吸收法（PI）和真菌粘附试验，证实了SK对烟曲霉（A. fumigatus）的抑制作用。与0.1% DMSO治疗组相比，SK治疗组的临床评分和IL-6、TNF-α、IL-1β水平均显著降低。SK还促进了自噬蛋白LC3B- II/I和Beclin-1的活性，同时降低了P62水平。SK处理后，RAW 264.7细胞中自溶酶体增加，真菌和分生孢子的吞噬作用增强。3-甲基腺嘌呤（3-MA）预处理抵消了SK降低炎症标志物的能力。此外，SK抑制烟曲霉生长，破坏细胞膜，破坏生物膜，抑制分生孢子粘附。综上所述，SK通过激活自噬具有抗炎功能，通过减少真菌负荷具有抗真菌功能，突出了其对真菌性角膜炎的潜在治疗作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Experimental eye research 医学-眼科学

CiteScore

6.80

自引率

5.90%

发文量

323

审稿时长

66 days

期刊介绍： The primary goal of Experimental Eye Research is to publish original research papers on all aspects of experimental biology of the eye and ocular tissues that seek to define the mechanisms of normal function and/or disease. Studies of ocular tissues that encompass the disciplines of cell biology, developmental biology, genetics, molecular biology, physiology, biochemistry, biophysics, immunology or microbiology are most welcomed. Manuscripts that are purely clinical or in a surgical area of ophthalmology are not appropriate for submission to Experimental Eye Research and if received will be returned without review.