Camila Camponogara1, Cássia R Silva2, Indiara Brusco1, Mariana Piana3, Henrique Faccin4, Leandro Machado de Carvalho4, André Schuch5, Gabriela Trevisan6, Sara Marchesan Oliveira7. 1. Laboratory Neurotoxicity and Psychopharmacology, Graduate Program in Biological Sciences: Toxicological Biochemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil. 2. Graduate Program in Genetics and Biochemistry, Institute of Genetics and Biochemistry, Federal University of Uberlandia, Uberlandia, MG, Brazil. 3. Phytochemical Research Laboratory, Graduate Program in Pharmaceutical Sciences, Health Sciences Center, Federal University of Santa Maria, Santa Maria, RS, Brazil. 4. Graduate Program in Chemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil. 5. Graduate Program in Biological Sciences: Toxicological Biochemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil. 6. Graduate Program in Physiology and Pharmacology, Federal University of Santa Maria, Santa Maria, Camobi, Santa Maria, RS, Brazil. 7. Laboratory Neurotoxicity and Psychopharmacology, Graduate Program in Biological Sciences: Toxicological Biochemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil; Graduate Program in Biological Sciences: Toxicological Biochemistry, Center of Natural and Exact Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil. Electronic address: saramarchesan@ufsm.br.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Inflammatory skin diseases treatments currently used cause adverse effects. Nasturtium officinale (watercress) is used popularly as an anti-inflammatory. However, until now, no study proved its effectiveness as a topical treatment to inflammatory skin diseases. The topical anti-inflammatory activity of N. officinale crude extract leaves (NoE) on an irritant contact dermatitis (ICD) model croton oil-induced in mice was investigated. MATERIALS AND METHODS: ICD models were induced by a single (1 mg/ear; acute) or repeated (0.4 mg/ear; chronic; 9 days total) croton oil application. NoE and dexamethasone solutions' (diluted in acetone; 20 μL/ear) or NoE gel, dexamethasone gel and base gel (15 mg/ear) were topically applied immediately after croton oil application. The NoE topical anti-inflammatory effect was evaluated for inflammatory parameters (ear edema, inflammatory cells infiltration, and inflammatory cytokines levels). NoE topical anti-inflammatory mechanism (NF-κB pathway and effect glucocorticoid-like) were assessed by western blot and ear edema analyses, respectively. UHPLC-MS/MS chromatography, gels accelerated stability and preliminary study of adverse effects was also performed. RESULTS: UHPLC-MS/MS of the NoE revealed the presence of coumaric acid, rutin, and ferulic acid. NoE gels stability study showed no relevant changes at low temperature. NoE, dexamethasone, NoE gel and dexamethasone gel inhibited the ear edema croton oil-induced by 82 ± 6% (1 mg/ear), 99 ± 1% (0.1 mg/ear), 81 ± 8% (3%) and 70 ± 6% (0.5%) for the acute model, and 49 ± 7% (1 mg/ear), 80 ± 4% (0.1 mg/ear), 41 ± 8% (3%) and 46 ± 14% (0.5%) for the chronic model, respectively. The same treatments also reduced the inflammatory cells infiltration by 62 ± 3% (1 mg/ear), 97 ± 2% (0.1 mg/ear), 60 ± 3% (3%) and 66 ± 6% (0.5%) for the acute model, respectively, and 25 ± 8% (1 mg/ear) to NoE and 83 ± 13% to dexamethasone to the chronic model. NoE and NoE gel reduced the pro-inflammatory cytokines levels (acute ICD model) by 62 ± 5% and 71 ± 3% (MIP-2) and 32 ± 3% and 44 ± 4% (IL-1β), while dexamethasone solution's and gel reduced by 79 ± 7% and 44 ± 4% to MIP-2 and 98 ± 2% and 83 ± 9% to IL-1β, respectively. NoE' and dexamethasone' solutions inhibited the reduction of IkB-α protein expression induced by croton oil by 100% and 80 ± 14%, respectively. Besides, the mifepristone (glucocorticoid receptor antagonist) pre-treatment prevented the topical anti-edematogenic effect of NoE' and dexamethasone' solutions by 61 ± 5% to NoE and 78 ± 16% to dexamethasone. The repeated topical application of NoE did not cause adverse effects. CONCLUSION: Our results suggest the N. officinale use in the cutaneous inflammatory process treatment and demonstrate the NoE potential to develop a promising topical anti-inflammatory agent to treat inflammatory disorders.
ETHNOPHARMACOLOGICAL RELEVANCE: Inflammatory skin diseases treatments currently used cause adverse effects. Nasturtium officinale (watercress) is used popularly as an anti-inflammatory. However, until now, no study proved its effectiveness as a topical treatment to inflammatory skin diseases. The topical anti-inflammatory activity of N. officinale crude extract leaves (NoE) on an irritant contact dermatitis (ICD) model croton oil-induced in mice was investigated. MATERIALS AND METHODS: ICD models were induced by a single (1 mg/ear; acute) or repeated (0.4 mg/ear; chronic; 9 days total) croton oil application. NoE and dexamethasone solutions' (diluted in acetone; 20 μL/ear) or NoE gel, dexamethasone gel and base gel (15 mg/ear) were topically applied immediately after croton oil application. The NoE topical anti-inflammatory effect was evaluated for inflammatory parameters (ear edema, inflammatory cells infiltration, and inflammatory cytokines levels). NoE topical anti-inflammatory mechanism (NF-κB pathway and effect glucocorticoid-like) were assessed by western blot and ear edema analyses, respectively. UHPLC-MS/MS chromatography, gels accelerated stability and preliminary study of adverse effects was also performed. RESULTS: UHPLC-MS/MS of the NoE revealed the presence of coumaric acid, rutin, and ferulic acid. NoE gels stability study showed no relevant changes at low temperature. NoE, dexamethasone, NoE gel and dexamethasone gel inhibited the ear edema croton oil-induced by 82 ± 6% (1 mg/ear), 99 ± 1% (0.1 mg/ear), 81 ± 8% (3%) and 70 ± 6% (0.5%) for the acute model, and 49 ± 7% (1 mg/ear), 80 ± 4% (0.1 mg/ear), 41 ± 8% (3%) and 46 ± 14% (0.5%) for the chronic model, respectively. The same treatments also reduced the inflammatory cells infiltration by 62 ± 3% (1 mg/ear), 97 ± 2% (0.1 mg/ear), 60 ± 3% (3%) and 66 ± 6% (0.5%) for the acute model, respectively, and 25 ± 8% (1 mg/ear) to NoE and 83 ± 13% to dexamethasone to the chronic model. NoE and NoE gel reduced the pro-inflammatory cytokines levels (acute ICD model) by 62 ± 5% and 71 ± 3% (MIP-2) and 32 ± 3% and 44 ± 4% (IL-1β), while dexamethasone solution's and gel reduced by 79 ± 7% and 44 ± 4% to MIP-2 and 98 ± 2% and 83 ± 9% to IL-1β, respectively. NoE' and dexamethasone' solutions inhibited the reduction of IkB-α protein expression induced by croton oil by 100% and 80 ± 14%, respectively. Besides, the mifepristone (glucocorticoid receptor antagonist) pre-treatment prevented the topical anti-edematogenic effect of NoE' and dexamethasone' solutions by 61 ± 5% to NoE and 78 ± 16% to dexamethasone. The repeated topical application of NoE did not cause adverse effects. CONCLUSION: Our results suggest the N. officinale use in the cutaneous inflammatory process treatment and demonstrate the NoE potential to develop a promising topical anti-inflammatory agent to treat inflammatory disorders.