Secondary metabolite contents and safety assessment study of the aqueous extract from the Algerian Echium trygorrhizum Pomel roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Species of Echium trygorrhizum Pomel, belonging to the Boraginaceae family, grow wild in North Africa. This plant is used in traditional Algerian medicine for the treatment of Diabetes, Jaundice and Tonsillitis. AIM OF THE STUDY: To our knowledge, no work has been done on the phytochemistry and biological activities of this plant. Moreover, no study has yet corroborated that the use of this plant is safe. Therefore, the present study was carried out to gather information on the various medicinal uses of this plant and to evaluate the total phenolic content and assess its safety after acute and sub-acute toxicity tests with rodents (mice and rats). MATERIAL AND METHODS: An ethnopharmacological survey was carried out using a direct questionnaire. It included some questions relating to the plant. The content of polyphénols, flavonoids and tannins present in the aqueous extract was determined by colorimetric methods. In the acute toxicity tests, three groups each containing five males and five females Albino mice were formed. The control group received water, while the test groups received different doses of aqueous extract (2000 and 5000) mg/kg body weight. In the subacute toxicity study, different doses (250, 500 and 1000 mg/kg) of the aqueous extract were administered to male and female rats for 28 consecutive days. Hematological analysis, biochemical parameters estimation and histopathological examination have been determined at the end of the treatment. RESULTS: Data from the ethnopharmacological survey showed that 25, 31% of people used this plant in Algeria as a traditional medicine for the treatment of Jaundice (100%), Diabetes (28, 33%) and Tonsillitis (10%). Results also revealed that aqueous extract contains high amounts of polyphenols, flavonoids and tannins. The extract did not cause any toxicity during the 14-days observation period after a single dose of 5000 mg/kg was administered to mice, with LD50 values greater than 5000 mg/kg. In a further step, the administration of the aqueous extract at all dose levels (250, 500, and 1000 mg/kg) to male and female rats during the 28-days study did not result in any deaths or behavioral changes, and there were no changes in body weight, relative organ weights, or food consumption as a result of this experiment. However, male rats showed a significant difference in relative liver weight in the high dose group (1000 mg/kg) (p < 0.05) and the satellite group (p < 0.01). In female rats, liver weight significantly increased only in the satellite group compared with the control group (p < 0.01). The results showed an increase in red blood cells (RBC), hemoglobin (Hb), and hematocrit (Hct) in the female groups. However, a clear decrease was observed in plasma ALP enzyme activities in females treated with the doses of 250 and 1000 mg/kg (p < 0.05) after the treatment periods. CONCLUSION: The study indicated that the single dose of 5 g/kg AQE can be considered relatively safe as it did not cause death or any signs of toxicity in mice. Repeated oral administration of AQE at doses below 250 g/kg/day for 28 consecutive days can be considered relatively safe.

Phytochemical Screening, In vitro Antioxidant and Enzyme Inhibitory Properties, and Acute Toxicity of Extracts from the Aerial Parts of Ephedra nebrodensis, a Source of Bioactive Compounds.

BACKGROUND: Due to the strong association between the chemistry of medicinal plants and their biological properties, it is important to determine their phytochemical composition to justify experimental tests. OBJECTIVE: The aim of this study was to evaluate the in vitro antioxidant and the enzyme inhibitory properties and to identify the bioactive compounds present in the extracts of Ephedra nebrodensis growing in Algeria. METHODS: Total phenolic and flavonoids content in these extracts were quantified by Folin- Ciocalteu and aluminum chloride methods. The antioxidant capacity was assessed using DPPH, ABTS, ß-carotene/linoleic acid, CUPRAC and FRAP assays, and in vitro cholinesterase activity against acetylcholinesterase and butyrylcholinesterase were evaluated. The chemical constituents of the extracts were analyzed by high-performance liquid chromatography coupled with mass spectrometric detection and gas chromatography. For the acute toxicity study, extracts were administered to mice at single dose of 2 g/kg and 5 g/kg by gavage. RESULTS: Plant extracts were rich in phenolic compounds. Ethyl acetate extract presented the highest phenolic (238.44 ± 1.50 µg GAE /mg of extract) and flavonoid (21.12 ± 0.00 µg QE /mg of extract) contents. Likewise, ethyl acetate extract showed potent radical scavenging and reducing properties. Ethanol-acetone extract showed inhibitory activity against acetylcholinesterase, and was a potent inhibitor of butyrylcholinesterase. In all extracts, flavonoids were the most abundant compounds. The phytochemical investigation showed the presence of alkaloids (ephedrine and pseudo-ephedrine). In the acute toxicity, the LD50 was superior to 5 g/kg body weight. There were no alterations in the histology of the liver and kidneys. CONCLUSION: This study demonstrated a good antioxidant potential and anticholinesterase activity of aerial parts of E. nebrodensis.

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.

In-vivo anti-inflammatory activity and safety assessment of the aqueous extract of Algerian Erica arborea L. (Ericaceae) aerial parts.

ETHNOPHARMACOLOGICAL RELEVANCE: Erica arborea known as Khlenj in Algeria is a small shrub belonging to Ericaceae family. E. arborea Aqueous extract (EAAE) is used in traditional medicine for anti-inflammatory, diuretic, antimicrobial, and antiulcer purposes. AIM OF THE STUDY: To our knowledge, no data reveal the combination between in-vivo anti-inflammatory and toxicological studies of EAAE. For this purpose, the aim of this study is to evaluate the biological activity cited above and assess its safety. MATERIAL AND METHODS: Anti-inflammatory activity was undergone using carrageenan-induced paw edema and croton oil-induced ear edema. The acute and sub-acute toxicity were conducted following the OECD guidelines 423 and 407, respectively. Phytochemical identification was carried out using HPLC-DAD-MS. Quantitative evaluation of polyphenols; flavonoids and antioxidant activity of EAAE were also determined. RESULTS: Oral administration of EAAE (250 and 500 mg/kg) significantly (p < 0.05) reduced the edema induced by carrageenan. Administration of EAAE dosed at 250 and 500 mg/kg exhibited efficacy in reducing edema induced by croton oil. The acute administration of EAAE at doses of 2000 and 5000 mg/kg did not cause any mortality or adverse effects indicating that the LD50 is above 5000 mg/kg. The prolonged administration of EAAE (500 and 1000 mg/kg) showed a significant reduction in triglycerides levels in male and female rats whereas no significant changes in other biochemical and hematological parameters were observed. Histopathological damages were recorded in both liver and kidney animal's tissues of both sexes treated with medium and maximum doses of EAAE. Phytochemical characterization of EAAE revealed a high amount of phenolic compounds, HPLC-DAD-MS analysis led to the identification of chlorogenic acid and five flavonol glycosides: myricetin pentoside, quercetin-3-O-glucoside, myricetin-3-O-rhamnoside, quercetin-3-O-pentoside, and quercetin-3-O-rhamnoside. CONCLUSION: In the light of the results obtained in this study, EAAE corroborates the popular use to treat the anti-inflammatory impairments. EAAE can be considered as non-toxic in acute administration and exhibited a moderate toxicity in sub-acute administration. High phenolic content and in-vitro antioxidant activity observed indicate that EAAE may reduce oxidative stress markers in-vivo.

Antioxidant and Anti-inflammatory Properties of Algerian Thymelaea microphylla Coss. and Dur. Extracts.

BACKGROUND: Thymelaea microphylla Coss. et Dur. (Thymelaeaceae) (TM) is a rare medicinal plant endemic to Algeria. Leaves decoction is used in folk medicine for anticancer, anti-inflammatory, and antidiabetic properties. OBJECTIVE: Herein, the antioxidant and anti-inflammatory properties of different extracts from leaves and flowers of Algerian TM were evaluated. MATERIALS AND METHODS: The study was carried out by in vitro cell-free assays (antioxidant/radical properties), ex vivo experiments (inhibition of prostaglandin E2 and thromboxane B2 release in human whole blood) and in vitro experiments on cell systems (cytotoxicity on peripheral blood mononuclear cells, and protective effects on human vein endothelial cells exposed to TNF-α). RESULTS: The acetone TM extract showed significant antioxidant properties and excellent anti-inflammatory and cyclooxygenase-inhibitory activity, together with lack of toxicity on normal human blood cells; furthermore, it was able to protect endothelial cells against dysfunction induced by TNF-α, as shown by decrease in cell death, e-selectin expression and leukocyte adhesion. CONCLUSION: On these bases, TM leaves and flowers appear to be a good source of bioactive compounds with significant antioxidant and antiinflammatory capability, and potentially effective in prevention and treatment of pathological conditions related to oxidative stress and inflammation, such as endothelial dysfunction. SUMMARY: Thymelaea microphylla leaves and flowers are a good source of bioactive compounds with significant antioxidant/free radical scavenger and antiinflammatory activity.The acetone extract from leaves and flowers of Algerian Thymelaea microphylla possesses excellent cyclooxygenase-inhibitory activity.This extract is able to protect against endothelial dysfunction, an early event in development of atherosclerosis and vascular diseases. Abbreviations used: TM: Thymelaea microphylla; BCB: Beta-carotene bleaching; AcE: Acetone extract; PGE2: Prostaglandin E2; TxB2: Thromboxane B2; FL: Fluorescein; Cat: Catechin; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azinobis-(3-ethyl-benzothiazolin-6-sulfonic acid)+; Que: Quercetin; ORAC: Oxygen radical absorbance capacity; AAPH: 2,2'-azobis (2-methylpropionamidine)dihydrochloride; PMS/NADH: Phenazine methosulfate/nicotinamide adenine dinucleotide; HUVECs: Human umbilical vein endothelial cells.

Flavonoid profile, antioxidant and cytotoxic activity of different extracts from Algerian Rhamnus alaternus L. bark.

BACKGROUND: Rhamnus alaternus (Rhamnaceae) L. has been traditionally used for treatment of many diseases. OBJECTIVE: In this study, we determined the antioxidant/free radical scavenger properties, the flavonoid profile and the cytotoxicity of aqueous and methanolic extracts obtained by maceration from Algerian R. alaternus bark, like also of aqueous extract prepared by decoction according to the traditional method. This to estimate the usefulness of the drug traditional preparation and compare it with those made in the laboratory. MATERIALS AND METHODS: The antioxidant activity of the extracts was evaluated using five different redox-based assays, all involving one redox reaction with the oxidant. High-performance liquid chromatography/diode array detection/electrospray ionization mass spectrometry analysis was used to identify and quantify the flavonoids content. Cytotoxicity on human monocytic leukemia cells (U937) was also carried out. RESULTS: All the extracts tested showed a good antioxidant/free radical scavenger activity and a similar flavonoid fingerprint. However, the methanolic one presented the best antioxidant activity that can be due to the highest flavonoid amount and significantly reduced the proliferation of leukemia cells. The results confirm that the extract prepared by decoction contains efficient antioxidant compounds and this justifies in part the therapeutic and preventive usefulness. Moreover, the methanolic extract exerted excellent cytotoxicity on U937 that could be attributed to kaempferol and rhamnocitrin glycosides.

Phenotypic and genotypic characterization of some lactic Acid bacteria isolated from bee pollen: a preliminary study.

In the present work, five hundred and sixty-seven isolates of lactic acid bacteria were recovered from raw bee pollen grains. All isolates were screened for their antagonistic activity against both Gram-positive and Gram-negative pathogenic bacteria. Neutralized supernatants of 54 lactic acid bacteria (LAB) cultures from 216 active isolates inhibited the growth of indicator bacteria. They were phenotypically characterized, based on the fermentation of 39 carbohydrates. Using the simple matching coefficient and unweighted pair group algorithm with arithmetic averages (UPGMA), seven clusters with other two members were defined at the 79% similarity level. The following species were characterized: Lactobacillus plantarum, Lactobacillus fermentum, Lactococcus lactis, Pediococcus acidilactici, Pediococcus pentosaceus, and unidentified lactobacilli. Phenotypic characteristics of major and minor clusters were also identified. Partial sequencing of the 16S rRNA gene of representative isolates from each cluster was performed, and ten strains were assigned to seven species: Lactobacillus plantarum, Lactobacillus fermentum, Lactococcus lactis, Lactobacillus ingluviei, Pediococcus pentosaceus, Lactobacillus acidipiscis and Weissella cibaria. The molecular method used failed to determine the exact taxonomic status of BH0900 and AH3133.

Endophytic fungi isolated from wheat (Triticum durum Desf.): evaluation of their antimicrobial activity, antioxidant activity and host growth promotion.

The emergence of antibiotic-resistant micro-organisms calls for inventive research and development strategies. The screening for antimicrobial compounds from endophytes is a promising way to meet the increasing threat of drug-resistant strains of human and plant pathogens. Endophytes may be defined as "microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects". Endophytes are relatively unstudied as potential sources of novel natural products for exploitation in medicine, agriculture, and industry. The purpose of this study was to evaluate several isolated fungi from wheat (Triticum durum Desf.) Mohamed Ben Bachir variety and to select endophytic fungi for further evaluation of its antimicrobial, antioxidant activities and host growth promotion. A total of 20 endophytic fungi have been isolated. Antimicrobial activity was evaluated for crude ethyl acetate extracts using an agar diffusion assay. All extracts showed inhibitory activity on at least one or more pathogenic microorganism, with an average zone of inhibition varied between 7 mm to 25 mm, a large zone of 23 and 25mm against candida albicans and Escherichia coli respectively. The antioxidant capacity of the extracts was evaluated by beta-carotene/linoleic acid assay. Results showed that 70% of these extracts have antioxidant activity, exhibiting 50, 57% to 78, 96% inhibitions. While 30% from them, their inhibitory activity for oxidation of linoleic acid Were less than 50%. Growth promotion ability of these endophytes was tested on seed germination among ten isolates tested, two isolates showed significant growth promotion effects on wheat seeds. From the present work we can conclude that these microorganisms could be promising source of bioactive compounds, growth promotion and warrant further study.