Leguminosae Lectins as Biological Tools in Medical Research: a Review

Abstract Lectins were discovered first in plants and later in other living things, and nowadays it is known that they are present in almost all many life forms. These proteins can bind to specific carbohydrates, which make them perform a number of biological activities and can be used as tools in the study of glycoconjugate structures present on the cell surface, being effective in medical research. Plant lectins, leguminosae lectins particularly, are among the most studied plant proteins. They are very versatile molecules, which show several interesting biological properties. Thus, the present paper reviewed the advances about the leguminosae lectins biological properties studies in the last ten years, taking into account their possible applications in the fields of Clinical Microbiology, Pharmacy and Cancerology through a search in the electronic databases, providing opportunity to exchange information about the theme. Leguminosae lectins can neutralize pathogenic organisms, be they viruses, prokaryotes and eukaryotes, in addition carcinogenic cells, besides decreasing oxidative stress, conditions which increases the possibility of alternative substances for the design of new drugs to be used in current therapeutic, expanding the possibilities of diseases cure.

Avaliação dos Efeitos Cardíacos de Lectina Solúvel em Água (WSMoL) de Sementes de Moringa Oleifera / Evaluation of the Cardiac Effects of a Water-Soluble Lectin (Wsmol) from Moringa Oleifera Seeds

Resumo Fundsamento As sementes de Moringa oleifera , que são utilizadas para clarificação de água, contêm uma lectina chamada WSMoL que tem mostrado atividade antibacteriana e imunomoduladora in vitro . Devido ao seu valor nutritivo e potencial terapêutico, as folhas e as sementes dessa árvore são consumidas em algumas comunidades. Algumas lectinas de plantas não são tóxicas para mamíferos, mas tem sido relatado que outras são prejudiciais quando ingeridas ou administradas por outros meios. Objetivo Como um dos passos necessários para determinar a segurança de WSMoL, nós avaliamos os possíveis efeitos cardiotóxicos desta proteína purificada. Métodos Durante 21 dias consecutivos, a WSMoL foi administrada a camundongos por gavagem. Foram investigadas as funções eletrofisiológicas, mecânicas e metabólicas in vivo e ex vivo por meio de registros eletrocardiográficos, ressonância magnética nuclear e respirometria de alta resolução. Resultados O tratamento com WSMoL não induziu alterações nos níveis de glicose no sangue ou peso corporal em comparação com o grupo controle. Adicionalmente, as relações peso cardíaco/peso corporal e peso cardíaco/comprimento tibial estavam semelhantes em ambos os grupos. A ingestão de lectina também não modificou a tolerância à glicose ou resistência à insulina. Não foram observadas alterações nos parâmetros eletrocardiográficos ou na duração do potencial de ação cardíaco. Os corações dos camundongos dos grupos controle e WSMoL mostraram função ventricular esquerda preservada. Além disso, a WSMoL não induziu alterações na função mitocondrial (em todos os casos, p > 0,05). Conclusões A administração de WSMoL demonstrou ter um perfil de segurança cardíaca. Estes resultados contribuem à avaliação de segurança do uso de sementes de M. oleifera para tratar água, visto que essa lectina está presente na preparação empregada por algumas populações com esse fim. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Abstract Background Moringa oleifera seeds, which are used for water clarification, contain a lectin named WSMoL which has shown in vitro antibacterial and immunomodulatory activity. Due to their nutritional value and therapeutic potential, the leaves and seeds of this tree are eaten in some communities. Some plant lectins are non-toxic to mammals, but others have been reported to be harmful when ingested or administered by other means. Objective As one of the steps needed to define the safety of WSMoL, we evaluated possible cardiotoxic effects of this purified protein. Methods: WSMoL was administered for 21 consecutive days to mice by gavage. Electrophysiological, mechanical, and metabolic cardiac functions were investigated by in vivo and ex vivo electrocardiographic recordings, nuclear magnetic resonance, and high-resolution respirometry. Results The treatment with WSMoL did not induce changes in blood glucose levels or body weight in comparison with control group. Moreover, the heart weight/body weight and heart weight/tibia length ratios were similar in both groups. Lectin ingestion also did not modify glucose tolerance or insulin resistance. No alterations were observed in electrocardiographic parameters or cardiac action potential duration. The heart of mice from the control and WSMoL groups showed preserved left ventricular function. Furthermore, WSMoL did not induce changes in mitochondrial function (in all cases, p > 0.05). Conclusions The administration of WSMoL demonstrated a cardiac safety profile. These results contribute to the safety evaluation of using M. oleifera seeds to treat water, since this lectin is present in the preparation employed by some populations to this end. (Arq Bras Cardiol. 2020; [online].ahead print, PP.0-0)

Effect of ScLL and 15d-PGJ2 on viability and cytokine release in LPS-stimulated fibroblasts: an in vitro study

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.

Effect of lectin (ScLL) on fibroblasts stimulated with LPS - an in vitro study

Abstract: The lectin (ScLL) extracted from the Synadenium carinatum plant has been evaluated as an immunomodulator in diseases such as asthma, neosporosis and leishmaniasis. However, it has not yet been evaluated in the oral cavity. This study evaluated the effect of ScLL on viability, proliferation and release of IL-10 in human gingival fibroblasts (HGF) stimulated with lipopolysaccharide (LPS). HGF were stimulated with LPS 1 µg/ml and treated with ScLL in concentrations of 10, 5 and 2 µg/ml for 1 and 5 h, and evaluated by flow cytometry for viability, apoptosis (initial/advanced) and necrosis. The supernatant was collected to detect release of IL-10 by ELISA. The proliferation was assessed with the BrdU assay. Positive control consisted of cells maintained in Dulbecco's Modified Eagles Medium (DMEM), and the negative control, of those kept in tap water. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found for ScLL concentrations regarding viability or initial and advanced apoptosis (p=0.455). All the groups, including the positive control, had a significantly lower necrosis parameter than negative control at 5 h (p < 0.001). No difference was found for proliferation among the experimental groups (p = 0.832). ScLL at 5 and 2 µg/ml resulted in a lower release of IL-10 than positive and negative controls at 5 h (p = 0.047). The results indicated that ScLL concentrations tested were not cytotoxic, and had no effect on proliferation and release of IL-10 parameters. A thorough understanding of ScLL, regarding its immunomodulatory potential, may open the door to new perspectives for dentistry.

Purification of a novel chitin-binding lectin with antimicrobial and antibiofilm activities from a Bangladeshi cultivar of potato (Solanum tuberosum).

A new chitin-binding lectin was purified from a Bangladeshi cultivar ‘Deshi’ of potato (Solanum tuberosum L.) through anion-exchange and affinity chromatographies using a chitin column. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the molecular mass of the lectin as 20,000 Daltons. This molecular mass was almost half of the molecular masses of chitin-binding lectins derived from other potatoes. The lectin showed both bactericidal and growth-inhibiting activities against Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli, Salmonella enteritidis and Shigella boydii) pathogenic bacteria. It also showed antifungal activity against Rhizopus spp., Penicillium spp. and Aspergillus niger. Biofilm produced by the bacterium Pseudomonas aeruginosa was dose-dependently reduced by 5-20% in 24 h after administration of the lectin, which was attributed to the glycan-binding property of the lectin having affinity to GlcNAc polymers. It was the first observation that any potato lectin prevented biofilm formation by P. aeruginosa and, therefore, could have possible applications in clinical microbiology and biomedical science.

Antinociceptive properties in mice of a lectin isolated from the marine alga Amansia multifida Lamouroux

The antinociceptive effects of a lectin (LEC) isolated from the marine alga Amansia multifida were determined in Swiss mice. The LEC (1, 5, and 10 mg/kg) inhibited acetic acid-induced abdominal writhings in a dose-dependent manner after intraperitoneal or oral administration. A partial but significant inhibition of writhings was observed after the combination of LEC (10 mg/kg) with avidin (1 mg/kg), a potent inhibitor of the hemmaglutinant activity of the lectin. However, total writhing inhibition was demonstrable in the group of mice treated with LEC plus mannose (1 mg/kg), as compared to LEC alone or to control groups. Furthermore, avidin and mainly mannose also play a role in antinociception, somehow facilitating the interaction of LEC with its active cell sites. In the formalin test, although both phases of the response were significantly inhibited, the effect of LEC was predominant during phase 2, causing inhibition of licking time that ranged from 48 to 88 percent after oral (5 and 10 mg/kg) and intraperitoneal (1 to 5 mg/kg) administration. As is the case with morphine, the effect of LEC (2 mg/kg) was reversed by naloxone (2 mg/kg), indicating the involvement of the opioid system. LEC was also effective in the hot-plate test, producing inhibitory responses to the thermal stimulus, and its effects were blocked by naloxone. In the pentobarbital-induced sleeping time, although LEC did not alter the onset of sleep significantly, it increased the time of sleep within the same dose range compared to control. These results show that LEC presents antinociceptive effects of both central and peripheral origin, possibly involving the participation of the opioid system.

Differential effect of plant lectins on mast cells of different origins

Histamine release induced by plant lectins was studied with emphasis on the carbohydrate specificity, external calcium requirement, metal binding sites, and mast cell heterogeneity and on the importance of antibodies bound to the mast cell membrane to the lectin effect. Peritoneal mast cells were obtained by direct lavage of the rat peritoneal cavity and guinea pig intestine and hamster cheek pouch mast cells were obtained by dispersion with collagenase type IA. Histamine release was induced with concanavalin A (Con A), lectins from Canavalia brasiliensis, mannose-specific Cymbosema roseum, Maackia amurensis, Parkia platycephala, Triticum vulgaris (WGA), and demetallized Con A and C. brasiliensis, using 1-300 æg/ml lectin concentrations applied to Wistar rat peritoneal mast cells, peaking on 26.9, 21.0, 29.1, 24.9, 17.2, 10.7, 19.9, and 41.5 percent, respectively. This effect was inhibited in the absence of extracellular calcium. The lectins were also active on hamster cheek pouch mast cells (except demetallized Con A) and on Rowett nude rat (animal free of immunoglobulins) peritoneal mast cells (except for mannose-specific C. roseum, P. platycephala and WGA). No effect was observed in guinea pig intestine mast cells. Glucose-saturated Con A and C. brasiliensis also released histamine from Wistar rat peritoneal mast cells. These results suggest that histamine release induced by lectins is influenced by the heterogeneity of mast cells and depends on extracellular calcium. The results also suggest that this histamine release might occur by alternative mechanisms, because the usual mechanism of lectins is related to their binding properties to metals from which depend the binding to sugars, which would be their sites to bind to immunoglobulins. In the present study, we show that the histamine release by lectins was also induced by demetallized lectins and by sugar-saturated lectins (which would avoid their binding to other sugars). Additionally, the lectins also released histamine from Rowett nude mast cells that are free of immunoglobulins.