Antioxidant and anti-inflammatory potentials of Ammodaucus leucotrichus Coss. & Durieu seeds' extracts: In vitro and in vivo studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Ammodaucus leucotrichus Coss. & Durieu (Apiaceae) is traditionally used in southern Algeria as a remedy against a wide range of disease due to its health-promoting properties. AIM OF THE STUDY: To investigate anti-oxidant and anti-inflammatory potentials of plant methanolic extract and its fractions in vitro and in vivo. MATERIALS AND METHODS: Anti-radical activity was assessed in vitro using ABTS•+, superoxide anion (O2•-) and nitric oxide radical (•NO). Lipid peroxidation inhibition was also investigated in the linoleic acid system. Enzyme inhibition assay was performed against α-amylase and α-glucosidase. The anti-inflammatory effect of extracts was screened in vitro through thermal induction of human serum albumin, and in vivo on a skin acute inflammation model induced by λ-carrageenan paw injection, xylene and croton oil topical application. Analgesic effect was evaluated by acetic acid-induced writhing test. RESULTS: The highest contents of polyphenols and flavonoids was recorded by the crude extract (77.14 ± 0.01 µg GAE/mg E and 19.59 ± 0.08 µg QE/mg E, respectively). Among the extracts, ethyl acetate extract showed a promising anti-radical activity of ABTS•+, O2•- and •NO, in addition to a remarkable inhibition activity of the tested enzymes. Meanwhile, all extracts effectively protected linoleic acid against lipid peroxidation and human serum albumin structure in thermal condition even at low concentration (0.31 mg/ml). Oral administration of 200 mg/kg of crude extract successfully inhibited acetic acid induced nociception and reduced edema formation induced by xylene and carrageenan. However, a dose-dependent manner was observed to decrease ear edema by a microscopic examination in croton oil induced acute inflammation. Nitrite and malondialdehyde levels together with catalase activity were modulated in the presence of plant-derived bioactive compounds. CONCLUSIONS: This study showed that Ammodaucus leucotrichus is potentially rich source of anti-oxidant and anti-inflammatory bioactive compounds.

In vitro Anti-hemolytic Effect, in vivo Anti-inflammatory and in vitro Anti-oxidant Activity of Anchusa azurea Mill.

BACKGROUND: Anchusa azurea Mill. (AA) is a medicinal plant largely used traditionally in folk medicine in Algeria; it is locally named hamham. It is effective in the treatment of various diseases. OBJECTIVES: The aim of the present study is to determine the antioxidant, anti-inflammatory, and anti- hemolytic effects of phenolic fractions from Anchusa azurea Mill. METHODS: In this study, various extracts from Anchusa azurea Mill. (AA) using solvents with increasing polarity were prepared. The quantification of polyphenols and flavonoids was determined. The anti-radical activity of the different extracts was evaluated using DPPH and by measuring the inhibition of the oxidative degradation of ß-carotene. The In-vitro antihemolytic effect of the plant extracts is determined (CrE, ChE, AcE, and AqE). For each extract, four concentrations were tested: 10.59, 21.18, 42.37, 84.74 µg/ml. Vitamin C is used as a standard. The free-radical attack was measured by measuring the HT50 (Half-Hemolysis Time). The anti-inflammatory effect using PMA on mice of the methanolic extract (CrE) was evaluated. RESULTS: The quantification of polyphenols and flavonoids showed that ethyl acetate extract (AcE) contains a higher amount of polyphenols. However, chloroform extract (ChE) presents a higher amount of flavonoids. AcE showed an important scavenging activity using the DPPH radical (IC50= 68.35 µg/ml). The results showed that AcE also exhibited a significant inhibition effect on the oxidation of ß-carotene/linoleic acid (84.33 %). All extracts increased the HT50 values (Half-Hemolysis Time) in a dose-dependent manner. The three highest concentrations (21.18, 42.37, and 84.74 µg / ml) of ChE caused a very significant delay (p ≤ 0.001) of hemolysis compared to the negative control and the positive control "VIT C". The anti-inflammatory effect of using PMA on mice showed that the methanolic extract (CrE) of AA reduced the weight of the ear edema. CONCLUSION: This plant has a strong pharmacological power, which supports its traditional medicinal use.

Antioxidant, Anti-inflammatory, and Analgesic Activities of Alcoholic Extracts of Ephedra nebrodensis From Eastern Algeria

Objectives: Ephedra nebrodensis (Ephedraceae) presents a wide range of biological activities. It is used to treat respiratory problems and hepatic pathologies in traditional medicine. The aim of this study is to evaluate the antioxidant, in vitro and in vivo anti-inflammatory and analgesic properties of two hydro-alcoholic extracts of E. nebrodensis in mice. Materials and Methods: The antioxidant capacity of hydro-methanolic (HM) and hydro-ethanolic (HE) extracts of E. nebrodensis was evaluated via assays of their superoxide radical scavenging capacity and ferrous ion chelating activity. The in vitro anti-inflammatory activity of the extracts (5, 10, and 20 mg/kg) was also determined using the bovine serum albumin denaturation test. Croton oil-induced ear edema was then employed to evaluate the in vivo anti-inflammatory effect of the extracts (200 and 400 mg/kg). Finally, the analgesic activity of the extracts (200 and 400 mg/kg) was determined by the acetic acid-induced torsion test. Results: The hydro-alcoholic extracts of E. nebrodensis present significant antioxidant activity. The HE and HM could inhibit protein denaturation by 82.99%±20.21% and 56.25%±2.12%, respectively. The extracts (HM and HE) also show strong anti-inflammatory effects in vivo and could reduce ear edema by 70.37%±2.00% and 72.22%±1.94%, respectively. The HM extract (72.51%±2.43%) demonstrates greater pain inhibitory effects than HE (70.76%±2.58%). Conclusion: The hydro-alcoholic extracts of E. nebrodensis produce antioxidant, anti-inflammatory, and analgesic effects. These results confirm the traditional use of the herb in the treatment of various diseases.

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.

Antioxidant and anti-inflammatory effect of Asphodelus microcarpus methanolic extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Asphodelus microcarpus is an important medicinal plant belonging to family Liliaceae. This plant is used in traditional medicine to treat abscesses by local application of the powder; the roots are used against white spots, and specifically used for ear pain. AIM OF THE STUDY: The aim of the present study was to assess the in vitro antioxidant effects and the in vivo anti-inflammatory activity of the different parts methanolic extracts of the Asphodelus microcarpus: aerial part (APME), leaf (LME), stem flowers (SFME) and root (RME). MATERIAL AND METHODS: The antioxidant potency of extracts was evaluated by DPPH (2, 2-diphenyl-l-picrylhydrazyl), ABTS, ß-carotene bleaching assays, iron chelating, free hydroxyl radicals (HO•) assays, and reducing power. The anti-inflammatory effect of the extracts was evaluated using the carrageenan induced rat paw edema and xylene-induced ear edema in mice. RESULTS: The estimation of polyphenols and flavonoids showed that the leave methanolic extract contains a high amount of polyphenols and flavonoids: 755.3 ±â€¯0.036 mg Gallic acid equivalent and 42.2 ±â€¯0.043 mg Rutin equivalent/g of dried material, respectively. Oral administration of the APME and RME extract produced significant (p < 0.05) anti-edematogenic effect with a dose of 500 mg/kg in the carrageenan induced paw edema after 6 h (58.04%, 58.75%, respectively). APME, LME; SFME and RME extracts at 100, 300 and 500 mg/kg, exhibited significant (p < 0.05) inhibition of xylene induced ear edema. CONCLUSION: the present study confirms the use of A. microcarpus in traditional medicine as anti-inflammatory agent.

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.

INVESTIGATION OF ANTIHEMOLYTIC, XANTHINE OXIDASE INHIBITION, ANTIOXIDANT AND ANTIMICROBIAL PROPERTIES OF SALVIA VERBENACA L. AERIAL PART EXTRACTS.

BACKGROUND: In this study, Salvia verbenaca L. aerial part extracts (SVEs): were screened for their antihemolytic, xanthine oxidase (XO) inhibition, antioxidant and antimicrobial activities. MATERIAL AND METHODS: To investigate SVEs antihemolytic activity, the 2,2,-azobis (2-amidinopropane) dihydrochloride (AAPH) was used to induce erythrocyte oxidative hemolysis. In XO inhibition test, xanthine was used as substrate and cytochrome c for generating superoxide anions. The antioxidant activity of SVEs was examined by means of reducing power, DPPH free radical scavenging and iron chelating assays. In addition, SVEs were tested for their antimicrobial effects by evaluating antibacterial and antifungal activities. RESULTS: Ethyl acetate extract (EAE) contains the highest amount of total polyphenols and flavonoids (661.78 ± 4.00 mg GAE / g E) and (28.81 ± 0.38 mg QE / g E) respectively. In antihemolytic test EAE was the most active extract with an HT50 value of 165 min. SVEs gave significant inhibitory effects on XO, especially the chloroform extract (ChE) with IC50 value of 0.0088 ± 0.000 mg/ml. EAE was the most active extract in reducing power essay (EC50: 0.0047 ± 0.000 mg/ml) and in DPPH radical scavenging essay (IC50: 0.0086 ± 0.000 mg/ml). Finally, the EAE has inhibited the growth of nine bacterial strains with inhibition zone diameters of (12 to 16 mm), but no activities have found against fungal strains. CONCLUSION: S. verbenaca could be considered as a potential source of natural antihemolytic, enzyme modulator, antioxidant and antibacterial agents.

Chromatographic fractionation, antioxidant and antibacterial activities of Urginea maritima methanolic extract.

The present work concerns a phytochemical study of Urginea maritima L. from Algeria, and an evaluation of antioxidant activity of the methanolic extract (UMME) and its chromatographic fractions. UMME was fractionated using open glass chromatography on silica gel and antioxidant effects were evaluated using DPPH and ß-carotene/linoleate assays. The phytochemical screening revealed that the bulb of plant contains flavonoids, glycosides, tannins, reducing compounds, anthraquinones combined, anthocyanins, mucilage, triterpenes and steroids. DPPH method showed that the UMME has a scavenger effect on radical DPPH with an IC50=57.83±1.59µg/ml. The fractions isolated from U. maritima (L.) presented an IC50 ranging between 499.23 and 39.68µg/ml. In ß-carotene/linoleate test, UMME and fractions give an I% =69.56±0.08% and between 31.29±0.49% and 90.79±0.29%, respectively. UMME showed a high inhibitory effect on the xanthine oxidase (IC50=0.67±0.01 mg/ml) and on the cytochrome c reduction (IC50=0.68 mg/ml). Wide range of phytochemical constituents in Urginea maritima were detected in methanolic extract which exhibited antioxidant and antibacterial activity. This plant could serve as pilot for the development of novel agents for pathological disorders.

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.

Free radical scanvenging and antioxidant effects of some anthraquinone derivatives.

In this study, the screening of five anthraquinones (purpurin, xanthopurpurin, rubiadin, kermisic acid and flavokermisic acid), for their free radical scavenging and antioxidant effects was carried out, using three complementary methods. DPPH (2,2'-diphenyl-1-picrylhydrazyl) revealed that purpurin has a scavenging effect with IC50 = 3.491 ± 0.014 µg/ml. Results of ß-carotene/linoleic acid assay showed that kermisic and flavokermisic acids have significant inhibition of lipid peroxidation with I % = 76.1 ± 1.5% and 68.6 ± 2.5%, respectively. In addition, the ferrous ion chelating test showed that only purpurin, with small concentrations, interferes in a dose dependant manner with the formation of Fe2+-ferrozine complex. These results are promising for further studies of the biological and pathological effects of these natural products.

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.