Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Lodz, Muszyńskiego 1, 90-151, Lodz, Poland.
Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236, Lodz, Poland.
J Ethnopharmacol. 2024 Jan 30;319(Pt 2):117219. doi: 10.1016/j.jep.2023.117219. Epub 2023 Sep 22.
Gaultheria procumbens L. is a polyphenolic-rich medicinal and food plant. Its leaves, stems, and fruits are traditional anti-inflammatory, antipyretic, antioxidant, and antimicrobial herbal medicines used to treat internal and external inflammation-related ailments, including rheumatic diseases, influenza, the common cold, fever, and skin and periodontal problems. Moreover, G. procumbens leaf extract is used for skin care as an anti-ageing and anti-wrinkle ingredient.
Various environmental factors, especially solar ultraviolet radiation, accelerate skin ageing by promoting oxidative stress and inflammation. Despite the dermoprotective and anti-ageing applications, the impact of G. procumbens on human dermal fibroblasts is unknown. Therefore, the study aimed to evaluate the anti-inflammatory, antioxidant, and photoprotective activity of G. procumbens standardised leaf, stem, and fruit extracts in cellular models, including human dermal fibroblasts (Hs68 cells) under UVA-irradiation, the primary pro-ageing skin stressor.
Hs68 fibroblasts were pre-treated (24h) with G. procumbens extracts (0.5-100 μg/mL) or reference compounds followed by UVA-irradiation (8 J/cm). Cell viability and metabolic activity were measured by CCK-8 and MTT assays in human Hs68 and mouse L929 fibroblasts, respectively. The ROS level, SOD, and GST activities were estimated by fluorescence and spectrophotometric techniques. The pro-inflammatory potential (NF-κB transcription factor activation) was checked using THP1-Blue™ NF-κB cells, and the anti-inflammatory activity was studied by measuring IL-8, ICAM-1, and NF-κB levels and phosphorylation of Erk kinase in LPS-stimulated Hs68 cells by spectrophotometry and confocal microscopy. The UVA-induced DNA damage and cell migration were evaluated by comet and scratch assays, respectively.
The extracts did not affect the metabolic activity of mouse L929 fibroblasts and the viability of unirradiated human Hs68 cells. Additionally, the extracts noticeably enhanced the viability of UVA-irradiated Hs68 cells up to 115-120% (p < 0.001) for stem and leaf extract at 25 μg/mL. All extracts in a wide concentration range (0.5-100 μg/mL) did not activate monocytes or induce the NF-κB transcription factor in LPS-stimulated Hs68 fibroblasts. On the other hand, the extracts (5-25 μg/mL) restored the activity of endogenous antioxidant enzymes, i.e., SOD and GST, up to 120-140% (p < 0.001) in the UVA-irradiated Hs68 cells. Moreover, a statistically significant reduction of ROS, IL-8, ICAM-1, and NF-κB levels by up to 48%, 88%, 43%, and 39%, respectively (p < 0.001) and strong suppression of Erk kinase activation was observed for the extracts (25-50 μg/mL) in LPS-stimulated human fibroblasts. The total DNA damage (% tail DNA) in irradiated Hs68 cells was also strongly decreased by up to 66-69% (p < 0.001) at 50 μg/mL. However, the treatment with the extracts did not relevantly enhance the cell migration of Hs68 fibroblasts.
The results suggest that G. procumbens may effectively protect human skin fibroblast from UVA irradiation. The leaf and stem extracts were the most potent antioxidants, while fruit and stem extracts revealed the strongest anti-inflammatory activity. The observed effects support the traditional use of aerial plant parts (leaves, stems, and fruits) in treating inflammation-related skin disorders cross-linked with oxidative stress and the topical application of Gaultheria extracts as anti-ageing agents intended for skin care.
Gaultheria procumbens L. 是一种多酚含量丰富的药用和食用植物。其叶子、茎和果实是传统的抗炎、解热、抗氧化和抗菌草药，用于治疗与炎症相关的内部和外部疾病，包括风湿性疾病、流感、普通感冒、发烧以及皮肤和牙周问题。此外，G. procumbens 叶提取物还可用于皮肤护理，作为抗衰老和抗皱成分。
各种环境因素，特别是太阳紫外线辐射，通过促进氧化应激和炎症加速皮肤衰老。尽管具有皮肤保护和抗衰老作用，但人们对 G. procumbens 对人类真皮成纤维细胞的影响知之甚少。因此，本研究旨在评估 G. procumbens 标准化叶、茎和果实提取物在细胞模型中的抗炎、抗氧化和光保护活性，包括在原发性皮肤衰老应激物 UVA 辐射下的人真皮成纤维细胞（Hs68 细胞）。
用 G. procumbens 提取物（0.5-100μg/mL）或参考化合物对 Hs68 成纤维细胞（24h）进行预处理，然后用 UVA 照射（8J/cm）。用 CCK-8 和 MTT 测定法分别在人 Hs68 和小鼠 L929 成纤维细胞中测定细胞活力和代谢活性。通过荧光和分光光度技术评估 ROS 水平、SOD 和 GST 活性。使用 THP1-Blue™NF-κB 细胞检查促炎潜力（NF-κB 转录因子激活），通过分光光度法和共聚焦显微镜测量 LPS 刺激的 Hs68 细胞中 IL-8、ICAM-1 和 NF-κB 水平以及 Erk 激酶磷酸化来研究抗炎活性。通过彗星和划痕试验分别评估 UVA 诱导的 DNA 损伤和细胞迁移。
提取物不影响小鼠 L929 成纤维细胞的代谢活性和未照射的人 Hs68 细胞的活力。此外，提取物显著提高了 UVA 照射的 Hs68 细胞的活力，最高可达 115-120%（p<0.001），在 25μg/mL 时为茎和叶提取物。所有提取物在广泛的浓度范围内（0.5-100μg/mL）都不会激活单核细胞或诱导 LPS 刺激的 Hs68 成纤维细胞中的 NF-κB 转录因子。另一方面，提取物（5-25μg/mL）将内源性抗氧化酶的活性提高至 120-140%（p<0.001），在 UVA 照射的 Hs68 细胞中为 SOD 和 GST。此外，在 LPS 刺激的人成纤维细胞中，提取物（25-50μg/mL）还观察到 ROS、IL-8、ICAM-1 和 NF-κB 水平分别降低 48%、88%、43%和 39%（p<0.001），Erk 激酶激活也得到强烈抑制。照射的 Hs68 细胞中的总 DNA 损伤（%尾巴 DNA）也降低了 66-69%（p<0.001），在 50μg/mL 时达到最高。然而，提取物处理并没有显著增强 Hs68 成纤维细胞的迁移。
结果表明，G. procumbens 可能有效地保护人皮肤成纤维细胞免受 UVA 辐射。叶和茎提取物是最有效的抗氧化剂，而果实和茎提取物则表现出最强的抗炎活性。观察到的效果支持传统上使用植物的地上部分（叶子、茎和果实）来治疗与炎症相关的皮肤疾病，这些疾病与氧化应激有关，并将 Gaultheria 提取物作为抗衰老剂应用于皮肤护理，以达到抗衰老的目的。
