Maria Inês Rocha1, Maria José Gonçalves2, Carlos Cavaleiro2, Maria Teresa Cruz3, Cláudia Pereira4, Patrícia Moreira3, Lígia Salgueiro2, Artur Figueirinha5. 1. University of Coimbra, Faculty of Sciences and Technology of University of Coimbra, Coimbra, Portugal; University of Coimbra, Faculty of Pharmacy of University of Coimbra, Coimbra, Portugal. 2. University of Coimbra, Faculty of Pharmacy of University of Coimbra, Coimbra, Portugal; CIEPQPF, Research Center for Chemical Processes Engineering and Forest Products, University of Coimbra, Portugal. 3. University of Coimbra, Faculty of Pharmacy of University of Coimbra, Coimbra, Portugal; CNC, Centre for Neuroscience and Cell Biology, University of Coimbra, Portugal. 4. CNC, Centre for Neuroscience and Cell Biology, University of Coimbra, Portugal; CIBB-Center for Innovative Biomedicine and Biotechnology, University of Coimbra, Rua Larga, Faculty of Medicine, Polo I, 1(st)floor, 3004-504 Coimbra, Portugal; Faculty of Medicine, Azinhaga de Santa Comba, Celas, 3000-548, Coimbra, Portugal. 5. University of Coimbra, Faculty of Pharmacy of University of Coimbra, Coimbra, Portugal; LAQV, REQUIMTE, Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal. Electronic address: amfigueirinha@ff.uc.pt.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia campestris L. is widely used in traditional medicine for their anti-inflammatory, antirheumatic, antimicrobial and antioxidant properties. A. campestris subsp. maritima Arcang., a halophyte plant ("madorneira" or "erva-lombrigueira" in Portugal), is traditionally used for gastric disorders, rheumatism and hypertension. AIM OF THE STUDY: The present study aims to characterize the essential oil (EO) and the hydrodistillation residual water (HRW), a by-product of the EO production, of Artemisia campestris subsp. maritima from Portugal and evaluate the antioxidant, antifungal, anti-inflammatory and wound healing activities of both extracts at concentrations without toxicity. MATERIALS AND METHODS: The phenolic profile of HRW was determined by HPLC-PDA-ESI/MSn and the EO was analyzed by gas chromatography (GC-FID and GC-MS). The antioxidant activity of both extracts were determined by several assays (ABTS, NO FRAP, β-carotene and DPPH). The antifungal activity (MIC and MLC) was evaluated against yeasts, dermatophytes and Aspergillus strains using broth macrodilution methods. The anti-inflammatory potential was determined measuring the nitric oxide (NO) production by stimulated mouse leukemic macrophages (RAW 264.7). Cell viability was evaluated for RAW 264.7 and mouse fibroblasts (NIH/3T3). The wound healing activity was evaluated in mouse fibroblasts (NIH/3T3) by the scratch assay. RESULTS: The HRW is mainly characterized by hydroxycinnamic acids and the essential oil is characterized by high amounts of β-pinene (54.5%), cadin-4-en-7-ol (9.5%), Z-β-ocimene (6.0%) and Ƴ-terpinene (4.6%). Both extracts showed high antioxidant potential in different assays. Additionally, only the hydrodistillation residual water showed anti-inflammatory activity (IC50 of 330 μg/mL). On the other hand, only the EO showed antifungal activity, particularly against Epidermophyton floccosum (MIC and MLC values of 0.16 μL/mL), and wound healing activity. Bothe extracts were not cytotoxic to macrophages CONCLUSIONS: The by-product HRW contains safe bioactive compounds with antioxidant and anti-inflammatory effect and the EO shows antioxidant properties, antifungal activity against dermatophytes and wound healing effect in skin cells. Overall, our results support the interest and economic value of two extracts obtained from a Portuguese native species and provide scientific validation to some of its traditional uses.
ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia campestris L. is widely used in traditional medicine for their anti-inflammatory, antirheumatic, antimicrobial and antioxidant properties. A. campestris subsp. maritima Arcang., a halophyte plant ("madorneira" or "erva-lombrigueira" in Portugal), is traditionally used for gastric disorders, rheumatism and hypertension. AIM OF THE STUDY: The present study aims to characterize the essential oil (EO) and the hydrodistillation residual water (HRW), a by-product of the EO production, of Artemisia campestris subsp. maritima from Portugal and evaluate the antioxidant, antifungal, anti-inflammatory and wound healing activities of both extracts at concentrations without toxicity. MATERIALS AND METHODS: The phenolic profile of HRW was determined by HPLC-PDA-ESI/MSn and the EO was analyzed by gas chromatography (GC-FID and GC-MS). The antioxidant activity of both extracts were determined by several assays (ABTS, NO FRAP, β-carotene and DPPH). The antifungal activity (MIC and MLC) was evaluated against yeasts, dermatophytes and Aspergillus strains using broth macrodilution methods. The anti-inflammatory potential was determined measuring the nitric oxide (NO) production by stimulated mouse leukemic macrophages (RAW 264.7). Cell viability was evaluated for RAW 264.7 and mouse fibroblasts (NIH/3T3). The wound healing activity was evaluated in mouse fibroblasts (NIH/3T3) by the scratch assay. RESULTS: The HRW is mainly characterized by hydroxycinnamic acids and the essential oil is characterized by high amounts of β-pinene (54.5%), cadin-4-en-7-ol (9.5%), Z-β-ocimene (6.0%) and Ƴ-terpinene (4.6%). Both extracts showed high antioxidant potential in different assays. Additionally, only the hydrodistillation residual water showed anti-inflammatory activity (IC50 of 330 μg/mL). On the other hand, only the EO showed antifungal activity, particularly against Epidermophyton floccosum (MIC and MLC values of 0.16 μL/mL), and wound healing activity. Bothe extracts were not cytotoxic to macrophages CONCLUSIONS: The by-product HRW contains safe bioactive compounds with antioxidant and anti-inflammatory effect and the EO shows antioxidant properties, antifungal activity against dermatophytes and wound healing effect in skin cells. Overall, our results support the interest and economic value of two extracts obtained from a Portuguese native species and provide scientific validation to some of its traditional uses.