Effects of Hypericin on Cultured Primary Normal Human Dermal Fibroblasts Under Increased Oxidative Stress.

Ioanna A Anastasiou, Panagiotis Sarantis, Ioanna Eleftheriadou, Konstantinos N Tentolouris, Iordanis Mourouzis, Michalis V Karamouzis, Konstantinos Pantos, Nikolaos Tentolouris
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Abstract

Introduction: Wound healing is a dynamic process that begins with inflammation, proliferation, and cell migration of a variety of fibroblast cells. As a result, identifying possible compounds that may improve fibroblast cell wound healing capacity is crucial. Hypericin is a natural quinine that has been reported to possess a wide range of pharmacological profiles, including antioxidant and anti-inflammatory, activities. Herein we examined for the first time the effect of hypericin on normal human dermal fibroblasts (NHDFs) under oxidative stress.

Methods: NHDF were exposed to different concentrations of hypericin (0-20 μg/mL) for 24 h. For the oxidative stress evaluation, H2O2 was used as a stressor factor. Cell viability and proliferation levels were evaluated. Immunohistochemistry and flow cytometry were performed to assess cell apoptosis levels and with confocal microscopy we identified the mitochondrial superoxide production under oxidative stress and after the treatment with hypericin. Scratch assay was performed under oxidative stress to evaluate the efficacy of hypericin in wound closure. To gain an insight into the molecular mechanisms of hypericin bioactivity, we analyzed the relative expression levels of genes involved in oxidative response and in wound healing process.

Results: We found that the exposure of NHDF to hypericin under oxidative stress resulted in an increase in cell viability and ATP levels. We found a decrease in apoptosis and mitochondrial superoxide levels after treatment with hypericin. Moreover, treatment with hypericin reduced wound area and promoted wound closure. The levels of selected genes showed that hypericin upregulated the levels of antioxidants genes. Moreover, treatment with hypericin in wound under oxidative stress downregulated the levels of proinflammatory cytokines, and metalloproteinases; and upregulated transcription factors and extracellular matrix genes.

Conclusion: These findings indicated that hypericin possesses significant in vitro antioxidant activity on NHDF and provide new insights into its potential beneficial role in the management of diabetic ulcers.

金丝桃素对氧化应激下培养的原代正常人真皮成纤维细胞的影响。
创面愈合是一个动态的过程,开始于各种成纤维细胞的炎症、增殖和细胞迁移。因此,确定可能改善成纤维细胞伤口愈合能力的化合物是至关重要的。金丝桃素是一种天然奎宁,据报道具有广泛的药理作用,包括抗氧化和抗炎活性。本文首次研究了金丝桃素对氧化应激下正常人真皮成纤维细胞(ndfs)的影响。方法:将NHDF暴露于不同浓度金丝桃素(0 ~ 20 μg/mL)作用24 h。氧化应激评价采用H2O2作为应激因子。评估细胞活力和增殖水平。通过免疫组织化学和流式细胞术评估细胞凋亡水平,并通过共聚焦显微镜鉴定氧化应激和金丝桃素处理后线粒体超氧化物的产生。采用氧化应激下划痕法评价金丝桃素在伤口愈合中的作用。为了深入了解金丝桃素生物活性的分子机制,我们分析了参与氧化反应和伤口愈合过程的基因的相对表达水平。结果:我们发现,在氧化应激下,NHDF暴露于金丝桃素导致细胞活力和ATP水平增加。我们发现金丝桃素治疗后细胞凋亡和线粒体超氧化物水平降低。此外,金丝桃素治疗可减少伤口面积,促进伤口愈合。所选基因水平表明金丝桃素上调抗氧化基因水平。此外,金丝桃素可下调氧化应激创面的促炎细胞因子和金属蛋白酶水平;转录因子和细胞外基质基因的上调。结论:金丝桃素具有显著的体外抗氧化活性,为其在糖尿病溃疡治疗中的潜在有益作用提供了新的认识。
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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