Phenolic profile, safety assessment, and anti-inflammatory activity of Salvia verbenaca L.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia species are known to have anti-inflammatory properties, and are traditionally used for healing purposes. Salvia verbenaca is an Algerian plant used for healing wounds and ulcers. AIM OF THE STUDY: This work aims to assess the acute and subacute safety of S. verbenaca and its possible anti-inflammatory activity as a mechanism contributing to its traditional applications. MATERIALS AND METHODS: Lethal toxicity of S. verbenaca hydromethanolic extract was evaluated against Artemia salina larvae, while acute and subacute toxicity were orally tested on mice. The anti-inflammatory activity was screened ex vivo using membrane stabilization and in vivo using xylene induced ear edema as an acute inflammation model. The antiradical, reducing power and iron chelating activities of S. verbenaca were also investigated in vitro, and phenolic compounds were determined using UHPLC-DAD-ESI-MSn. RESULTS: Salvia verbenaca extract contained high amounts of phenolic compounds (206 µg GAE/mg extract). The in vitro antioxidant activity showed promising radical scavenging ability, iron chelating (IC50: 189 µg/mL), reducing power and strong anti-lipid-peroxidation effect (IC50: 111 µg/mL). The extract had potential cytotoxic effect against Artemia salina larvae (LC50: 30 µg/mL), but did not exhibit any acute/subacute toxicity effect on mice. Salvia verbenaca inhibited hypotonic and heat induced hemolysis and also reduced 50% of xylene induced ear edema at 600 mg/kg bw. Rosmarinic acid and caffeoylmalic acid were identified as the major compounds. CONCLUSION: Salvia verbenaca hydromethanolic extract was found to be safe at acute and subacute levels. Its in vitro/in vivo antioxidant activity, membrane stabilizing properties and anti-inflammatory activity may be an important aspect of its wound healing and anti-ulcer traditional use.

Thymus algeriensis Bioss & Reut: Relationship of phenolic compounds composition with in vitro/in vivo antioxidant and antibacterial activity.

Thymus algeriensis Bioss & Reut is an Algerian native plant traditionally used for culinary and medicinal purposes. To evaluate the in vivo antioxidant and antimicrobial properties of T. algeriensis, phenolic compounds were extracted using hydromethanolic solutions and administrated to Swiss albinos mice. It was observed that the plasma antioxidant capacity increased, as well as catalase and glutathione levels, whereas malondialdehyde decreased. In vitro assays confirmed that the extract scavenged 2,2-diphenyl-1-picrylhydrazyl, DPPH• (7 µg/mL), chelated (EC50: 512 µg/mL) and reduced iron ions (5.3 mM FeSO4/g), and inhibited ß-carotene bleaching (90% at 2 mg/mL). Antibacterial effects were also observed towards Salmonella typhimurium and Proteus mirabilis. However, the methanolic fraction obtained by reversed solid phase cartridge, showed antibacterial activity against Escherichia coli (MIC = 9.37 mg/mL), Proteus mirabilis (MIC = 4.68 mg/mL), Salmonella typhimurium (MIC = 7.06 mg/mL), Micrococcus luteus (MIC = 7.03 mg/mL), and Bacillus cereus (MIC = 2.34 mg/mL). UHPLC-DAD-ESI-MSn analysis showed that these properties could result from rosmarinic acid, caffeoyl rosmaniric acid, and kaempferol and eriodictyol glycoside derivatives. These results pave the way for the understanding of T. algeriensis traditional applications and its use for the development of novel food applications.

Acute, sub-acute toxicity and antioxidant activities (in vitro and in vivo) of Reichardia picroide crude extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Reichardia picroide is a species mainly used for alimentary purposes, but it is traditionally known to be used as hypoglycemiant, diuretic, depurative, galactagogue and tonic. AIM OF THE STUDY: To date, there are no studies corroborating both its safety and antioxidant activities. The objective of the present study, thus, was to assess the safety profile of Reichardia picroide methanolic extract (RPE) and as well as on its antioxidant and antihemolytic activities. MATERIALS AND METHODS: The acute toxicity of RPE was carried out based on OECD guidelines 425. Signs accompanying toxicity and possible death of animals were monitored for two weeks to ascertain the median lethal dose (LD50) of the RPE. In sub-acute toxicity study, the extract was administered by gavage at the doses of 250, 500 and 1000mg/kg/day for 21 consecutive days. The antioxidant activity of RPE was investigated through various methods both in vitro and in vivo. RESULTS: The admistrated doses did not produce mortality or changes in general behaviors of the tested males and females mice. The LD50 was found to be superior to 5000mg/kg DW. Moreover, daily administration of RPE at doses ranged from 500 to 1000mg/kg could result in alteration of liver and kidney histology. Significant decrease in liver enzymes (ALT and AST), urea and creatinine levels in female plasma was recorded. The RPE was, in vitro, strong in DPPH scavenging and hemolytic inhibition, benificial in lipid peroxidation inhibition and reducing power. In addition, it exhibited, in vivo, a strong effect on GSH level increasing and lipid peroxidation inhibition in liver and kidney. CONCLUSIONS: It can be suggested, based on the results of this study, that the crude extract of Reichardia picroide was non-toxic in acute administration and the use of this extract is safe at doses ≤ 250mg/kg. This study supports the application of Reichardia picroides in alimentary and traditional medicine purposes. Moreover, antioxidant activity results suggested that Reichardia picroide had potent antioxidant activities and could be utilized as new natural antioxidant in food and therapeutics.

Antioxidant Potentials and Xanthine Oxidase Inhibitory Effect of Two Furanocoumarins Isolated from Tamus communis L.

In this investigation, the screening of two furanocoumarins; 5,8- dimethoxypsoralen (1) and heraclinin (2), isolated from the methanol root-extracts of Tamus communis L for their antioxidant activity and xanthine oxidase inhibitory effect was carried out, using different assays such as DPPH free radical scavenging effect, ß- carotene / linoleic acid, xanthine oxidase (XO) inhibition and in addition to blood total antioxidant capacity. Results revealed that the two compounds have significant DPPH radical scavenging activity and effective inhibition of linoleic acid oxidation in a dose-dependent manner; 5,8-dimethoxypsoralen exhibited the highest activity with an I% = 72.69 ± 1.88%. These results indicate that the isolated compounds inhibit xanthine oxidase activity and scavenge superoxide radicals with heraclinin (2) as the more potent xanthine oxidase inhibitor, and 5,8-dimethoxypsoralen (1) as the more effective on cytochrome c reduction, the two tested compounds can effectively protect erythrocytes against hemolytic injury induced by AAPH. These results are promising for further studies of the biological and pathological effects of these natural products.

Comparison of xanthine oxidase levels in synovial fluid from patients with rheumatoid arthritis and other joint inflammations.

OBJECTIVE: To search whether xanthine oxido-reductase (XOR) present in the synovium is also liberated, to determine its activity in synovial fluid and to establish a possible relationship between XOR levels in rheumatoid arthritis (RA) and non-RA patients. METHODS: This study was carried out in the Laboratory of Immunology, University Ferhat Abbas, Setif, Algeria from 2001-2008. This study is a retrospective controlled study matching cases with RA to non rheumatoid joint inflammations. Synovial fluid (SF) samples were collected with consent of the patients, at Setif University Hospital, from adults suffering from RA (n=36) or only with joint inflammations (n=52). After its detection in SF with indirect enzyme-linked immunosorbent assay (ELISA) and dot-immunobinding, using anti-bovine XOR as first antibodies, XOR was assayed with capture ELISA. RESULTS: Xanthine oxidoreductase is found in all studied SF. Capture ELISA showed levels up to 0.762 and 0.143 mg/mL in SF of RA and other joint inflammations patients, respectively. In most cases, more than 50% of synovial XOR is present as oxidase form. Positive correlation was observed between enzyme level and the disease severity since RA patients had a significantly high enzyme amount compared to patients with other less severe arthritic pathologies. CONCLUSION: These results suggest that the enzyme could well be involved in joint inflammation probably by producing reactive oxygen species.

Antioxidant properties and xanthine oxidase inhibitory effects of Tamus communis L. root extracts.

This study was conducted to search for xanthine oxidase (XO) inhibitors from the root extracts of Tamus communis L. traditionally used in folk medicine in Algeria. Root extracts with different solvents were screened for purified milk xanthine oxidase inhibition. The root extracts (methanol, chloroform and ethyl acetate) and proteins, obtained in distilled water, inhibited bovine, sheep and human milk XO from three species in a concentration-dependent manner, with an additional superoxide scavenging capacity, which reached its highest level with ethyl acetate extract (IC(50) = 0.15, 0.04 and 0.09 g/L) for bovine XO, sheep XO and human XO, respectively. The antioxidant potential was confirmed with the non-enzymatic method, total radical-trapping antioxidant parameter (TRAP) assay, which showed that the Tamus communis L. extracts have a potential antioxidant activity in the same order obtained by using the reduction of cytochrome c, an enzymatic method, in which the antioxidant activity followed a decreasing order: ethyl acetate extract > chloroform extract > protein.