In Vitro Anti-Inflammatory Effect ofSalvia sagittataEthanolic Extract on Primary Cultures of Porcine Aortic Endothelial Cells

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.

Clinopodium tomentosum(Kunth) Govaerts Leaf Extract Influencesin vitroCell Proliferation and Angiogenesis on Primary Cultures of Porcine Aortic Endothelial Cells

“…Moreover, CTEE showed a good antioxidant activity, as a results from both DPPH (3.72 mmol TE/g) and ORAC (4.14 mmol TE/g) assays, as shown in Table 2. TPC in CTEE resulted very similar to that reported for a similar ethanolic extract of Salvia sagittata, a species belonging to the same family [34], and was of the same order of magnitude as that reported in a Thymus vulgaris methanolic extract [50].…”

“…Total flavonoid content (TFC) was determined according to Zhishen et al [41] with some modifications [34]. Results were expressed as mmol rutin equivalents (RE)/g extract.…”

“…The recovery values of phenolic acids in spiked samples ranged from 78.8 to 92.2% (RSD < 9:8%, n = 6). The chromatographic method for the analysis of flavonoids was adapted from Wojdyło et al [43], as previously reported [34]. Gradient elution was carried out with a mixture of 4.5% formic acid and acetonitrile.…”

Effects of Hydrogen Sulfide Donor NaHS on Porcine Vascular Wall-Mesenchymal Stem Cells

“…The pVW-MSCs were cultured for 21 days in the presence or absence of NaHS, at each change of the medium the NaHS treatment was repeated. After 21 days, the pVW-MSCs were lysed then the RNA extraction and cDNA synthesis were carried out as previously described [45]. The gene expression analysis of VW-MSC markers (thy-1 cell surface antigen (CD90), Nestin, neuron-glial antigen 2 (NG2), platelet-derived growth factor receptor β (PDGFRβ) and α-smooth muscle actin (αSMA)) was performed by quantitative real time PCR (qPCR) as previously reported by us [26].…”

“…The gene expression analysis of VW-MSC markers (thy-1 cell surface antigen (CD90), Nestin, neuron-glial antigen 2 (NG2), platelet-derived growth factor receptor β (PDGFRβ) and α-smooth muscle actin (αSMA)) was performed by quantitative real time PCR (qPCR) as previously reported by us [26]. The gene expression analysis was conducted as previously reported by using two different reference genes (β-Actin (β-Act) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)) [45]. Primer sequences, expected PCR product lengths and accession numbers are reported in Table 1.…”