Antioxidant, Antihemolytic, Antihyperuricemic, Anti-inflammatory Activity of Algerian Germander Methanolic Extract

The Germander (Teucrium polium) is commonly used as a medicinal plant in Algeria against a variety of human diseases. This study aims to investigate the antioxidant, anti-hemolytic and antihyperuricemic effects of the Algerian germander (Teucrium polium L.) extract. T. polium witch was collected from Bordj Bouarreridj, Algeria and extracted with methanol to give the methanolic extract (TPME). The objective of this work is to disassemble, at first, the antioxidant effect of TPME in vivo and in vitro, secondarily to evaluate the anti-inflammatory and finally to study for the first time the hypouricemic activity. The quantification of polyphenols and flavonoids showed that the TPME contains 160.72±0.78 µg EAG/mg of polyphenols and 37.96±0.317 µg EQ/mg of flavonoids. The antioxidant activities were carried out in mice by an in vivo assay, the plasma ability to inhibit DPPH radical and FRAP. TPME showed a protective effect against oxidative stress in erythrocytes. The total antioxidant defence system appears to be enhanced in the plasma, by increased FRAP levels probably due to higher levels of polyphenols in the Teucrium polium extract. The treated group showed an essential activity in the DPPH test compared to Vit C and control groups (28.64±5.84% vs 47.27±6.78% and 21.42±3.89%, respectively). The total antioxidant capacity of plasma and red blood cells was determined using the kinetics of hemolysis by the determination of HT50 (hemolysis half-life). The HT50 which was 179.6±10.53 min for treated group for, 158.2±3.85 for Vit C group and 146.5±1.78 min for the control, respectively. The present work demonstrated that Teucrium polium extract exerts a strong in vivo free radical scavenging and antioxidant activities. These activities are probably related to polyphenols and flavonoids. Hyperuricemia witch is induced by injection of potassium oxonate "PO", the uric acid, urea and creatinine were measured in plasma and supernatant of the liver. To evaluate their hypouricemic effect, TPME was administered intraperitoneally to potassium oxonate-induced hyperuricemic mice at a dose of 50 mg/kg body weight. TPME caused a decrease in plasma uric acid (3.3±0.18 mg/l) compared to control group (1.48±0.07 mg/l), almost the same value of uric acid of "PO" group. For "OP" group, value of uric acid in plasma is increased 4 times (6.33±1.22 mg/l) and almost 2 times for liver supernatant (31.36± 5.4 mg/l), the administration of 10 mg/kg of allopurinol decreased uric acid levels to normal (1.89±0.32 mg /l, 16.36±1.03 mg /l, respectively for plasma and supernatant). The findings data for the supernatant didn’t show any significant decrease in plasma and liver uric acid comparing the urea level of "OP" group (0.48 g/l); we can conclude that the rate of urea and creatinine after treatment with plant extract is normal and that the results of this study indicate the absence of renal damage in mice. The present study was undertaken to evaluate the anti-inflammatory effect of TPME in vivo The administration of TPME (100 mg/kg body wt.), reduced ear edema induced by phorbol myristate acetate (PMA), achieving a low degree of anti-inflammatory activity (%I = 18.46±1.59%), the effect was comparable with that of diclofenac used as a reference drug (%I = 38.84±1.87 %). The histopathological analysis indicated that the treatment with TPME led to a moderate decrease of the inflammatory infiltrate with a persistence of the oedema, against the injection of diclofenac, led to a significant reduction of the leucocytes. These results support the use of this plant in traditional medicine for inflammation disorder.

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 beta-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 IC[50]=57.83 +/- 1.59 micro g/ml. The fractions isolated from U. maritime [L.] presented an IC[50] ranging between 499.23 and 39.68 micro g/ml. In beta-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 [IC[50]=0.67 +/- 0.01 mg/ml] and on the cytochrome c reduction [IC[50]=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

Kinetics of Inhibition of Xanthine Oxidase by Globularia alypum and its Protective Effect against Oxonate-Induced Hyperuricemia and Renal Dysfunction in Mice.

In the present study, Globularia alypum (GA) was extracted with solvent of varying polarity allowed its separation into four subtractions: crude extract (CrE) chloroform extract (ChE), ethyl acetate extract (AcE) and aqueous extracts (AqE). The results showed that AcE had the highest content of phenolic compounds. The inhibitory activity of the extracts on xanthine oxidase (XO) was evaluated, the results obtained showed that the inhibition is dose-dependent and the AcE had a very significant inhibitory effect (0.069 ± 0.003 mg/ml) followed by CrE and AqE (0.081 ± 0.000 et 0.088 ± 0.002 mg/ml, respectively). The CrE, ChE and AqE inhibit competitively XO, whereas AcE is a non-competitive inhibitor. Hyperuricemia was induced by intraperitoneally injection of potassium oxonate, the uric acid, urea and creatinine were measured in serum and liver supernatant. The ChE, AcE, and AqE extracts had reduced significantly the plasma and liver uric acid levels (decreased 44.76%, 43.03% and 31.31%, respectively). Comparing these results with those obtained in vitro, the ChE extract with the lowest inhibitory effect in vitro (IC50 = 0,123 mg / ml) was the most effective extract in vivo. In summary, there was a contradiction between inhibition in vitro and in vivo, this inconsistency or difference may be due to the difference in the bioavailability of flavonoids or natural substances and their extensive metabolism in mice.

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

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. 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. 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. These results suggest that the enzyme could well be involved in joint inflammation probably by producing reactive oxygen species

Anti-xanthine oxidase antibodies in sera and synovial fluid of patients with rheumatoid arthritis and other joint inflammations

To study anti-bovine milk xanthine oxidoreductase XOR antibody levels in synovial fluid as well as in serum of patients suffering from rheumatoid affections to assess a possible correlation between antibody titres and severity of disease. Sera and synovial fluids were collected from volunteer donors at Setif University Hospital, Setif, Algeria from 2001-2007 with the consent of patients. Human IgG and IgM levels of free and bound anti-bovine milk XOR antibodies were determined using bovine XOR as antigen, with enzyme-linked immunosorbent assay ELISA. Serum IgG anti-bovine milk XOR titres in 30 healthy normal subjects 2.74 +/- 2.31 microgram/mL are in agreement with that reported in the literature. Immunoglobulin G and IgM anti-bovine milk XOR antibody titres were found to be significantly higher in serum from patients with rheumatoid arthritis RA, and latex positives subjects. Synovial IgM antibody titres to bovine XOR were found to be significantly higher in rheumatoid arthritis patients compared to patients with other joint inflammations. In rheumatoid arthritis patients, high concentrations of antibodies against XOR were noticed. These antibodies may play a major role in RA by inhibiting both xanthine and NADH oxidase activities of XOR. They may also play a key role in eliminating XOR from serum and synovial fluid positive role but unfortunately, immune complex formation could also activate complement and participate in self maintenance of inflammation