Anti-Inflammatory Effects of Supercritical Carbon Dioxide Extract and Its Isolated Carnosic Acid from Rosmarinus officinalis Leaves

Abstract

Rosemary (Rosmarinus officinalis) leaves possess a variety of bioactivities. Previous studies have shown that the extract of rosemary leaves from supercritical fluid extraction inhibits the expression of inflammatory mediators with apparent dose-dependent responses. In this study, three different extraction conditions (5000 psi at 40, 60, and 80 °C) of supercritical carbon dioxide (SC-CO(2)) toward the extraction of antioxidants from rosemary were investigated. Furthermore, simultaneous comparison of the anti-inflammatory properties between rosemary extract prepared from SC-CO(2) under optimal conditions (5,000 psi and 80 °C) and its purified carnosic acid (CA) using lipopolysaccharide (LPS)-treated murine RAW 264.7 macrophage cells was also presented. Results showed that the yield of 3.92% and total phenolics of 213.5 mg/g extract obtained from the most effective extraction conditions showed a high inhibitory effect on lipid peroxidation (IC(50) 33.4 μg/mL). Both the SC-CO(2) extract and CA markedly suppressed the LPS-induced production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), phosphorylated inhibitor-kappaB (P-IκB), and nuclear factor-kappaB (NF-κB)/p65 in a dose-dependent manner. The five major compounds of verbenone, cirsimaritin, salvigenin, carnosol, and CA existing in the SC-CO(2) extract were isolated by semipreparative HPLC and identified by HPLC-MS/MS analysis. CA was the most abundant recorded compound and the most important photochemical with an anti-inflammatory effect with an IC(50) of 22.5 μM or 7.47 μg/mL presented to the best inhibitory activity on NO production better than that of the 14.50 μg/mL dosage prepared from the SC-CO(2) extract. Nevertheless, the effective inhibition of LPS-induced NF-κB signaling in RAW 264.7 cells from the SC-CO(2) extract extends the potential application of nutraceutical formulation for the prevention of inflammatory diseases.

ObtainingAntioxidants from Botanic Matrices Applying Novel Extraction Techniques

“…The results obtained by Kuo et al [121] corroborate the studies carried out by Vicente et al [89] using rosemary leaves; the authors obtained high CA content in the extract in 1 h of extract ion by SFE. The antio xidant activity o f the extract increased with the ext raction time because the volatile oil is depleted from the vegetal matrix at the beginning of the process, and the phenolic co mpounds, which present higher antioxidant activity, are only later extracted.…”

“…Carnosic acid (CA) and carnosol are the major phenolic diterpenes present in rosemary ext racts obtained by SFE, as shown by Kuo et al [121]. The CA content obtained in the referred study was approximately 110 mg/g extract, resulting in IC 50 of 7.47 µg/cm 3 .…”

Antinociceptive effectiveness of triterpenes from rosemary in visceral nociception

“…The presence of hesperidin as one of the major constituents in an ethanol extract, which was shown to be partially responsible for the antinociceptive activity of R. officinalis assayed in an arthritic model in rats was previously described (Martínez et al, 2009b). Flavonoids and phenolic acids isolated from several medicinal plants are considered to be responsible for the analgesic, anti-inflammatory and anti-spasmodic activities in experimental models of abdominal, arthritic and orofacial pain (Martínez et al, 2009a;Bonjardim et al, 2011;Kuo et al, 2011). Moreover, anti-inflammatory properties of the triterpenes have also been described in several studies (Liu, 1995;Má ñ ez et al, 1997;Costa et al, 2003;Maia et al, 2006a;Altinier et al, 2007).…”

Effect of Rosmarinic acid on sertoli cells apoptosis and serum antioxidant levels in rats after exposure to electromagnetic fields

“…Rosmarinic acid (RA) is used to treat bronchial asthma, peptic ulcers, cataract, arthritis, cancer and rheumatoid arthritis. It is found in large quantities in oregano, lemon balm, sage, marjoram, and rosemary (Kuo et al, 2011). Many studies have shown that rosemary extracts play important roles in antiinflammation, anti-tumor, and anti-proliferation in various in vitro and in vivo settings (Cheng et al, 2011).…”