Abstract：
To determine the antifungal, anti-inflammatory and neuroprotective effects of resveratrol (RES) in Aspergillus fumigatus (A. fumigatus) keratitis.
Cytotoxicity assay and Draize eye assay were performed to assess the toxicity of RES. The antifungal effect of RES was assessed by minimal inhibitory concentration, scanning or transmission electron microscopy, propidium iodide uptake assay, and Calcofluor white staining. Phosphorylation of p38 MAPK, mRNA and protein levels of Dectin-1 and related inflammatory factors were measured by qRT-PCR, ELISA and Western blot in vitro and in vivo. Clinical score, HE staining, plate count, and myeloperoxidase test were used to observe the progress of fungal keratitis. IF staining, qRT-PCR, and the Von Frey test were selected to assess the neuroprotective effects of RES.
RES suppressed A. fumigatus hyphae growth and altered hyphae morphology in vitro. RES decreased the expression of Dectin-1, IL-1β and TNF-α, as well as p38 MAPK phosphorylation expression, and also decreased clinical scores, reduced inflammatory cell infiltration and neutrophil activity, and decreased fungal load. RES also protected corneal basal nerve fibers, down-regulated mechanosensitivity thresholds, and increased the mRNA levels of CGRP and TRPV-1..
These evidences revealed that RES could exert antifungal effects on A. fumigatus and ameliorate FK through suppressing the Dectin-1/p38 MAPK pathway to down-regulate IL-1β, IL-6, etc. expression and play protective effect on corneal nerves.
Cytotoxicity assay and Draize eye assay were performed to assess the toxicity of RES. The antifungal effect of RES was assessed by minimal inhibitory concentration, scanning or transmission electron microscopy, propidium iodide uptake assay, and Calcofluor white staining. Phosphorylation of p38 MAPK, mRNA and protein levels of Dectin-1 and related inflammatory factors were measured by qRT-PCR, ELISA and Western blot in vitro and in vivo. Clinical score, HE staining, plate count, and myeloperoxidase test were used to observe the progress of fungal keratitis. IF staining, qRT-PCR, and the Von Frey test were selected to assess the neuroprotective effects of RES.
RES suppressed A. fumigatus hyphae growth and altered hyphae morphology in vitro. RES decreased the expression of Dectin-1, IL-1β and TNF-α, as well as p38 MAPK phosphorylation expression, and also decreased clinical scores, reduced inflammatory cell infiltration and neutrophil activity, and decreased fungal load. RES also protected corneal basal nerve fibers, down-regulated mechanosensitivity thresholds, and increased the mRNA levels of CGRP and TRPV-1..
These evidences revealed that RES could exert antifungal effects on A. fumigatus and ameliorate FK through suppressing the Dectin-1/p38 MAPK pathway to down-regulate IL-1β, IL-6, etc. expression and play protective effect on corneal nerves.
摘要：
目的：为了确定抗真菌药，白藜芦醇（RES）在烟曲霉中的抗炎和神经保护作用（A.烟曲霉）角膜炎。
方法:进行细胞毒性测定和Draize眼测定以评估RES的毒性。通过最小抑制浓度评估RES的抗真菌作用，扫描或透射电子显微镜，碘化丙啶摄取测定，和钙氟白色染色。p38MAPK的磷酸化，qRT-PCR检测Dectin-1及相关炎症因子的mRNA和蛋白水平,体外和体内ELISA和Westernblot。临床评分,HE染色,平板计数,用髓过氧化物酶试验观察真菌性角膜炎的进展。IF染色,qRT-PCR,选择VonFrey试验来评估RES的神经保护作用。
结果：RES在体外抑制了烟曲霉菌丝的生长并改变了菌丝的形态。RES降低Dectin-1、IL-1β和TNF-α的表达，以及p38MAPK磷酸化表达，也降低了临床评分，减少炎症细胞浸润和中性粒细胞活性，减少真菌负荷。RES还保护角膜基底神经纤维，下调的机械敏感性阈值，并增加CGRP和TRPV-1的mRNA水平。.
结论：这些证据表明，RES可以通过抑制Dectin-1/p38MAPK通路下调IL-1β来发挥抗真菌作用，改善FK，IL-6等.对角膜神经的表达和发挥保护作用。
方法:进行细胞毒性测定和Draize眼测定以评估RES的毒性。通过最小抑制浓度评估RES的抗真菌作用，扫描或透射电子显微镜，碘化丙啶摄取测定，和钙氟白色染色。p38MAPK的磷酸化，qRT-PCR检测Dectin-1及相关炎症因子的mRNA和蛋白水平,体外和体内ELISA和Westernblot。临床评分,HE染色,平板计数,用髓过氧化物酶试验观察真菌性角膜炎的进展。IF染色,qRT-PCR,选择VonFrey试验来评估RES的神经保护作用。
结果：RES在体外抑制了烟曲霉菌丝的生长并改变了菌丝的形态。RES降低Dectin-1、IL-1β和TNF-α的表达，以及p38MAPK磷酸化表达，也降低了临床评分，减少炎症细胞浸润和中性粒细胞活性，减少真菌负荷。RES还保护角膜基底神经纤维，下调的机械敏感性阈值，并增加CGRP和TRPV-1的mRNA水平。.
结论：这些证据表明，RES可以通过抑制Dectin-1/p38MAPK通路下调IL-1β来发挥抗真菌作用，改善FK，IL-6等.对角膜神经的表达和发挥保护作用。
