Holothuria grisea agglutinin (HGA): the first invertebrate lectin with anti-inflammatory effects.

Holothuria grisea agglutinin (HGA) is a dimeric lectin of molecular mass 228 kDa by gel filtration with monomers of 105 kDa by SDS-PAGE. The lectin is highly thermostable as it retains full activity for 1 h at 70 °C. Unlike other lectins purified from marine invertebrates, the hemagglutination activity of HGA does not require any divalent metal ions. The affinity analysis of HGA showed that only mucin was able to inhibit the hemagglutinating activity. HGA administered intravenously was tested in classical models of nociception and inflammation. HGA was able to inhibit neutrophil migration into the peritoneal cavity induced by carrageenan. This inhibitory effect was 68% at a dose of 1 mg/kg. In acetic acid-induced writhing tests, a significant antinociceptive effect was observed by treatment with HGA (0.1; 1 or 10 mg/kg) reducing constrictions by 27, 90 and 84%, respectively. In formalin tests, HGA at a dose of 10 mg/kg showed antinociceptive effect only in the inflammatory phase (phase 2). Nevertheless, in hot-plate tests, HGA did not show any nociceptive effect. In rota-rod and open-field tests, HGA did not alter the animals' behavior. The treatment with HGA 10 mg/kg presented diminished myeloperoxidase activity activity (81.6% inhibition) and raised the circulating levels of NO by 50.4% when compared with the carrageenan group. HGA has demonstrated the ability to modulate the inflammatory response in models of inflammation in vivo. HGA is the first marine invertebrate lectin that showed an anti-inflammatory effect. This finding opens a new perspective on the potential of lectins from the marine environment.

In vivo anti-inflammatory effect of a sulfated polysaccharide isolated from the marine brown algae Lobophora variegata.

CONTEXT: Lobophora variegata J.V. Lamouroux (Dictyotaceae) is a brown marine alga widely encountered in the Brazilian sea coast that presents high content of fucans. Anti-inflammatory effects of fucans are reported mostly in models in vitro, but little is known about its effects in vivo. OBJECTIVE: To investigate vascular and cellular effects of a sulfated polysaccharide from the brown marine algae L. variegata (SP-Lv) in acute inflammatory models. MATERIALS AND METHODS: SP-Lv was isolated by DEAE-cellulose and analyzed by agarose gel electrophoresis and evaluated for its inhibitory effect on paw edema, vascular permeability, leukocyte migration and peritoneal nitrite content induced by zymosan in Wistar rats. Anticoagulant activities and possible systemic toxicity were also evaluated. RESULTS: SP-Lv inhibited the paw edema (120 min: 1.42 ± 0.11 vs. 0.95 ± 0.05 mL), plasma exudation (21.53 ± 0.62 vs. 11.96 ± 0.68 µg/g), nitrite content (4.42 ± 0.33 vs. 2.86 ± 0.003 µM) and leukocyte migration (5.15 ± 1.21 vs. 1.99 ± 0.16 cells/10(3) mL) induced by zymosan. SP-Lv and L-NAME reduced the paw edema (60-120 min) elicited by L-arginine. However, at 180 min SP-Lv effect was more accentuated and sustained until 240 min, while that of L-NAME was abolished. Similarly to indomethacin, SP-Lv inhibited the entire edema time-course induced by phospholipase A(2), except for the time of 60 min. DISCUSSION AND CONCLUSION: The anti-edematogenic effect of SP-Lv seems to occur via inhibition of nitric oxide synthase and cyclooxygenase activities. These results suggest a potential applicability of polysaccharides from alga origin in acute inflammatory conditions.

Antinociceptive and anti-inflammatory effects of a mucin-binding agglutinin isolated from the red marine alga Hypnea cervicornis.

The agglutinin from the red marine alga Hypnea cervicornis (HCA) was tested in models of nociception and inflammation. The role of carbohydrate-binding sites and the systemic toxicity were assessed. HCA (10(-1), 1, and 10 mg/kg) administered i.v. to mice inhibited writhes induced by acetic acid and, at 10 mg/kg, inhibited the second phase of the formalin test, but did not alter the response latency in the hot-plate test. HCA (1 mg/kg) administered i.v. to rats reduced carrageenan-induced paw edema at 1, 2, and 3 h after challenge, but not edema induced by dextran. The neutrophil migration induced by both N-formyl-methionyl-leucyl-phenylalanine (fMLP) and carrageenan was inhibited by HCA at 10(-1), 1, and 10 mg/kg. The combination of HCA (1 mg/kg) and its ligand mucin reversed the lectin inhibitory effect on carrageenan-induced neutrophil migration and acetic acid-induced writhes. The i.v. treatment of rats with HCA (1 mg/kg) for 7 days did not affect body mass; liver, kidney or heart wet weight; blood leukocyte counts; urea, creatinine or serum transaminase activity; or macroscopy of the organs examined. In short, H. cervicornis agglutinin showed important antinociceptive and anti-inflammatory activity via interaction with the lectin carbohydrate-binding site.

In vitro and in vivo antitumor effect of 5-FU combined with piplartine and piperine.

It has been reported that piplartine and piperine, alkaloid/amide compounds from Piper species, show antitumor activities. In the present paper, the effects of the combination of 5-fluorouracil (5-FU) with piplartine or piperine was determined using in vitro and in vivo experimental models. Hematological and biochemical analyses, as well as histopathological and morphological analyses of the tumor and the organs, including liver, spleen and kidney, were performed in order to evaluate the toxicological aspects associated with different treatments. The incubation of tumor cell lines with 5-FU in the presence of piplartine or piperine produced an increase in growth inhibition, as observed by lower IC50 values for 5-FU. These effects were also observed in vivo, where the combination with piplartine but not piperine with 5-FU led to a higher tumor growth inhibition. The results indicated that either piplartine- or 5-FU-treated animals showed a low inhibition rate when they were used individually at low doses of 28.67% and 47.71%, respectively, but when they were combined at the same dose, the inhibition rate increased significantly to 68.04%. The histopathological analysis showed that the livers and the kidneys of treated animals were only slightly and reversibly affected. Neither the enzymatic activity of transaminases nor the urea levels were significantly modified when compared with the control group. Hematological analysis showed leukopenia after 5-FU treatment, which was reversed by the combined use of piplartine and piperine. These findings indicate that piplartine may enhance the therapeutic effectiveness of chemotherapeutic drugs, and moreover, this combination could improve immunocompetence hampered by 5-FU.