Abstract
The aim of the present research was to study the effects of an ethanolic extract of Salvia sagittata Ruiz & Pav (SSEE), an endemic Ecuadorian plant traditionally used to treat inflammation and different intestinal affections, on primary cultures of porcine aortic endothelial cells (pAECs). pAECs were cultured in the presence of different concentrations (1-200 μg/mL) of SSEE for 24 h, and cytotoxicity was evaluated by the MTT assay. SSEE did not negatively affect cellular viability at any concentration tested. Cell cycle was analyzed and no significant change was observed. Then, the anti-inflammatory effects of SSEE on pAECs were analyzed using a lipopolysaccharide (LPS) as the inflammatory stimulus. Different markers involved in the inflammatory process, such as cytokines and protective molecules, were evaluated by real-time quantitative PCR and Western blot. SSEE showed the ability to restore pAEC physiological conditions reducing interleukin-6 and increasing Heme Oxygenase-1 protein levels. The phytochemical composition of SSEE was also evaluated via HPLC-DAD and spectrophotometric assays. The presence of different phenolic acids and flavonoids was revealed, with rosmarinic acid as the most abundant component. SSEE possesses an interesting antioxidant activity, as assessed through both the Oxygen Radical Absorbance Capacity (ORAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. In conclusion, results suggest that SSEE is endowed with an in vitro anti-inflammatory effect. This represents the initial step in finding a possible scientific support for the traditional therapeutic use of this plant.
Highlights
In the last few years, researches aimed to scientifically define the effects of natural products have been growing, due to the increasing popularity of plant-based Traditional Medicine and because it meets the primary health-care needs for the majority of the population in developing countries [1]
KG, Düren, Germany), and an iScript cDNA Synthesis Kit and iTaq Universal SYBR Green Supermix were used for cDNA synthesis and quantitative real-time PCR (qPCR) analysis, respectively (Bio-Rad Laboratories Inc., Hercules, CA, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Sigma-Aldrich
Folin-Ciocalteu’s phenol reagent, 1,1-diphenyl-2picrylhydrazyl (DPPH), 6-hydroxyl-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), 2,2-Azobis(2-methylpropionamidine) dihydrochloride (AAPH), fluorescein, gallic acid, rutin, phenolic acids (4-hydroxybenzoic, caffeic, chlorogenic, ferulic, gallic, p-coumaric, synapic, syringic, transcinnamic, and rosmarinic acids), quercetin, quercetin-3-O-glucoside, quercetin-3-O-rhamnoside, quercetin-3-O-galactoside, kaempferol, kaempferol-3-O-rutinoside, hesperetin, hesperidin pure standards (>99.5% purity) in powder form, and HPLC-grade solvents were purchased from Sigma-Aldrich
Summary
In the last few years, researches aimed to scientifically define the effects of natural products have been growing, due to the increasing popularity of plant-based Traditional Medicine and because it meets the primary health-care needs for the majority of the population in developing countries [1]. More than 20,000 plant species are used to treat several diseases and are considered as potential reservoirs for new drugs [2]. Recent studies suggest that the historical ethnopharmacological uses of plant-based medicines can represent a useful preliminary screening tool in the field of drug discovery [3]. The flora of mainland Ecuador is extremely rich: an estimated total of 17,000 species have so far been recorded [4, 5] and more than 3,000 medicinal plants are used in different native communities living on the highlands of the Ecuadorian Andes [6]. In most cases, the preparation, doses, and routes of administration of these herbal remedies are only
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