Skowrońska Weronika, Pawłowska Karolina A, Obrębski Maciej, Piwowarski Jakub P, Żyżyńska-Granica Barbara, Granica Sebastian
Microbiota Lab, Department of Pharmaceutical Microbiology and Bioanalysis, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland; Department of Pharmaceutical Biology, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
Microbiota Lab, Department of Pharmaceutical Microbiology and Bioanalysis, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
J Ethnopharmacol. 2025 Aug 15;353(Pt B):120433. doi: 10.1016/j.jep.2025.120433.
The bark of witch hazel (Hamamelis virginiana L.) has been employed in traditional medicine primarily due to its astringent properties, which are attributed to the tannins it contains. It has been utilized to alleviate the symptoms of haemorrhoids, inflammation of the skin and mucous membranes, and minor wounds. However, the potential changes in the chemical composition and biological activity of witch hazel bark extract following incubation with skin microbiota have not yet been analysed.
The objective of the study was to isolate and identify the principal components of the H. virginiana bark extract and monitor the alterations occurring during incubation with the skin microbiota ex vivo. Subsequently, the activity of the extract, its isolated compounds, and the primary metabolite was evaluated to determine their potential for alleviating skin cell inflammation, promoting wound healing, and exhibiting antibacterial effects.
The composition of the extract and the identification of metabolites following incubation with ex vivo skin microbiota were conducted using UHPLC-DAD-ESI-IT-MS. The principal components isolated by preparative HPLC were identified on the basis of their NMR spectra. Biological studies were conducted on human neutrophils ex vivo, stimulated with f-MLP (ROS production) and LPS (IL-1β, CXCL-8 and TNF-α secretion), as well as on HaCaT keratinocytes stimulated with TNF-α/IFN-γ and on fibroblasts (NHDF) stimulated with lipoteichoic acid (IL-6 and CXCL-8 release). The most potent samples were subjected to an in vitro wound healing assay, along with testing for their ability to inhibit the growth of Staphylococcus aureus.
The extract and hamamelitannin exhibited the most pronounced anti-inflammatory activity. Conversely, the principal metabolite generated during incubation with the skin microbiota, ellagic acid, was observed to stimulate the release of inflammatory mediators, particularly by fibroblasts. In the in vitro wound healing assay, both the extract and hamamelitannin supported migration of keratinocytes to the site of injury, with hamamelitannin demonstrating significantly stronger activity. The minimal inhibitory concentration for S. aureus growth was determined to be 0.5 mg/mL for the extract and 0.5 mM for hamamelitannin. Furthermore, both the extract and hamamelitannin were observed to prolong the lag phase and doubling time of the species.
The aqueous extract of Hamamelidis cortex, along with its key component, hamamelitannin, has been identified as a promising agent with anti-inflammatory and wound-healing properties. Nevertheless, the extract and its components may undergo chemical modifications due to the metabolism of the skin microbiota potentially leading to changes in biological activity with undesirable consequences.
北美金缕梅（Hamamelis virginiana L.）的树皮在传统医学中得到应用，主要是因其具有收敛特性，这归因于其所含的单宁。它已被用于缓解痔疮、皮肤和黏膜炎症以及轻微伤口的症状。然而，北美金缕梅树皮提取物与皮肤微生物群孵育后化学成分和生物活性的潜在变化尚未得到分析。
本研究的目的是分离和鉴定北美金缕梅树皮提取物的主要成分，并监测其在体外与皮肤微生物群孵育过程中发生的变化。随后，对提取物、其分离出的化合物以及主要代谢产物的活性进行评估，以确定它们在减轻皮肤细胞炎症、促进伤口愈合和发挥抗菌作用方面的潜力。
使用超高效液相色谱 - 二极管阵列 - 电喷雾 - 离子阱 - 质谱法对提取物的成分以及与体外皮肤微生物群孵育后的代谢产物进行鉴定。通过制备型高效液相色谱分离出的主要成分根据其核磁共振光谱进行鉴定。对体外用人源化甲酰甲硫氨酸 - 亮氨酸 - 苯丙氨酸（f - MLP）刺激的人中性粒细胞（活性氧生成）和脂多糖刺激的人中性粒细胞（白细胞介素 - 1β、趋化因子 - 8和肿瘤坏死因子 - α分泌）、用肿瘤坏死因子 - α/干扰素 - γ刺激的HaCaT角质形成细胞以及用脂磷壁酸刺激的成纤维细胞（NHDF）（白细胞介素 - 6和趋化因子 - 8释放）进行生物学研究。对最有效的样品进行体外伤口愈合试验，并测试它们抑制金黄色葡萄球菌生长的能力。
提取物和金缕梅鞣质表现出最显著的抗炎活性。相反，在与皮肤微生物群孵育过程中产生的主要代谢产物鞣花酸，被观察到会刺激炎症介质的释放，尤其是成纤维细胞释放炎症介质。在体外伤口愈合试验中，提取物和金缕梅鞣质均支持角质形成细胞向损伤部位迁移，金缕梅鞣质表现出明显更强的活性。提取物对金黄色葡萄球菌生长的最低抑菌浓度测定为0.5mg/mL，金缕梅鞣质为0.5mM。此外，提取物和金缕梅鞣质均被观察到会延长该菌种的迟缓期和倍增时间。
北美金缕梅树皮水提取物及其关键成分金缕梅鞣质已被确定为具有抗炎和伤口愈合特性的有前景的药物。然而，由于皮肤微生物群的代谢作用，提取物及其成分可能会发生化学修饰，这可能导致生物活性发生变化并产生不良后果。
