RESUMEN
Lectins are proteins widely distributed among plants, animals and microorganisms that have the ability to recognize and interact with specific carbohydrates. They have varied biological activities, such as the inhibition of the progression of infections caused by fungi, bacteria, viruses and protozoa, which is related to the interaction of these proteins with the carbohydrates present in the cell walls of these microorganisms. Leishmaniasis are a group of endemic infectious diseases caused by protozoa of the genus Leishmania. In vitro and in vivo tests with promastigotes and amastigotes of Leishmania demonstrated that lectins have the ability to interact with glycoconjugates present on the cell surface of the parasite, it prevents their development through various mechanisms of action, such as the production of ROS and alteration of membrane integrity, and can also interact with defense cells present in the human body, thus showing that these molecules can be considered alternative pharmacological targets for the treatment of leishmaniasis. The objective of the present work is to carry out a bibliographic review on lectins with leishmanicidal activity, emphasizing the advances and perspectives of research in this theme. Through the analysis of the selected studies, we were able to conclude that lectins have great potential for inhibiting the development of leishmaniasis. However, there are still few studies on this subject.
Asunto(s)
Lectinas , Leishmania , Leishmaniasis , Leishmania/efectos de los fármacos , Humanos , Lectinas/farmacología , Lectinas/química , Lectinas/metabolismo , Leishmaniasis/tratamiento farmacológico , Leishmaniasis/parasitología , Animales , Antiprotozoarios/farmacología , Antiprotozoarios/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
INTRODUCTION: Trypsin inhibitors (TIs) have the ability to competitively or non-competitively bind to trypsin and inhibit its action. These inhibitors are commonly found in plants and are used in protease inhibition studies involved in biochemical pathways of pharmacological interest. OBJECTIVES: This work aimed to purify a trypsin inhibitor from Bauhinia pulchella seeds (BpuTI), describing its kinetic mechanism and anticoagulant effect. METHODS: Affinity chromatography, protein assay, and SDS-PAGE were used to purify the inhibitor. Mass spectrometry, inhibition assays, and enzyme kinetics were used to characterize the inhibitor. In vitro assays were performed to verify its ability to prolong blood clotting time. RESULTS: Affinity chromatography on a Trypsin-Sepharose 4B column gave a yield of 43.1. BpuTI has an apparent molecular mass of 20 kDa with glycosylation (1.15%). Protein identification was determined by MS/MS, and BpuTI showed similarity to several Kunitz-type trypsin inhibitors. BpuTI inhibited bovine trypsin as an uncompetitive inhibitor with IC50 (3 x 10-6 M) and Ki (1.05 x 10-6 M). Additionally, BpuTI showed high stability to temperature and pH variations, maintaining its activity up to 100ºC and in extreme pH ranges. However, the inhibitor was susceptible to reducing agents, such as DTT, which completely abolished its activity. BpuTI showed an anticoagulant effect in vitro at a concentration of 33 µM, prolonging clotting time by 2.6 times. CONCLUSION: Our results suggest that BpuTI can be a biological tool to be used in blood clotting studies.
Asunto(s)
Bauhinia , Inhibidores de Tripsina , Animales , Bovinos , Inhibidores de Tripsina/farmacología , Inhibidores de Tripsina/química , Bauhinia/metabolismo , Tripsina/análisis , Tripsina/química , Tripsina/metabolismo , Espectrometría de Masas en Tándem , Semillas/química , Anticoagulantes/farmacología , Anticoagulantes/análisis , Anticoagulantes/químicaRESUMEN
Lectins are proteins of non-immunological origin with the ability to bind to carbohydrates reversibly. They emerge as an alternative to conventional antifungals, given the ability to interact with carbohydrates in the fungal cell wall inhibiting fungal growth. The lectin from D. violacea (DVL) already has its activity described as anti-candida in some species. Here, we observed the anti-candida effect of DVL on C. albicans, C. krusei and C. parapsilosis and its multiple mechanisms of action toward the yeasts. Additionally, it was observed that DVL induces membrane and cell wall damage and ROS overproduction. DVL was also able to cause an imbalance in the redox system of the cells, interact with ergosterol, inhibit ergosterol biosynthesis, and induce cytochrome c release from the mitochondrial membrane. These results endorse the potential application of DVL in developing a new antifungal drug to fight back against fungal resistance.
Asunto(s)
Dioclea , Lectinas , Lectinas/farmacología , Candida/metabolismo , Dioclea/metabolismo , Lectinas de Plantas/farmacología , Lectinas de Plantas/metabolismo , Antifúngicos/farmacología , Carbohidratos , Semillas/metabolismo , Ergosterol , Candida albicans , Pruebas de Sensibilidad MicrobianaRESUMEN
DVL is a Man/Glc-binding lectin from Dioclea violacea seeds that has the ability to interact with the antibiotic gentamicin. The present work aimed to evaluate whether the DVL has the ability to interact with neomycin via CRD and to examine the ability of this lectin to modulate the antibiotic effect of neomycin against multidrug-resistant strains (MDR). The hemagglutinating activity test revealed that neomycin inhibited the hemagglutinating activity of DVL with a minimum inhibitory concentration of 50 mM, indicating that the antibiotic interacts with DVL via the carbohydrate recognition domain (CRD). DVL immobilized on cyanogen bromide-activated Sepharose® 4B bound 41 % of the total neomycin applied to the column, indicating that the DVL-neomycin interaction is efficient for purification processes. Furthermore, the minimum inhibitory concentrations (MIC) obtained for DVL against all strains studied were not clinically relevant. However, when DVL was combined with neomycin, a significant increase in antibiotic activity was observed against S. aureus and P. aeruginosa. These results demonstrate the first report of lectin-neomycin interaction, indicating that immobilized DVL has the potential to isolate neomycin by affinity chromatography. Moreover, DVL increased the antibiotic activity of neomycin against MDR, suggesting that it is a potent adjuvant in the treatment of infectious diseases.
Asunto(s)
Dioclea , Fabaceae , Humanos , Masculino , Lectinas/farmacología , Antibacterianos/farmacología , Dioclea/química , Neomicina/farmacología , Lectinas de Plantas/química , Staphylococcus aureus/metabolismo , Fabaceae/metabolismoRESUMEN
The use of schiff base complex against microbial agentes a has recently received more attention as a strategy to combat infections caused by multidrug-resistant bacteria and leishmania. This study aimed to evaluate the toxicity, antibacterial and leishmanicidal activities of the nickel (II) chloride schiff base complex ([Ni(L2)] against Leishmania amazonensis promastigote, multi-resistant bacterial strains and evaluate to modulate antibiotic activity against multi-resistant bacterial. The schiff base complex was characterized by the techniques of elemental analysis, Fourier transform infrared spectroscopy (FTIR), UV-vis absorption spectroscopy and thermal analysis (TGA/DTG/DSC). The [Ni(L2)] complex presented moderate toxicity in saline artemia (LC50 = 150.8 µg/mL). In leishmanicidal assay, the NiL2 complex showed values of IC50 of (6.079 µg/mL ± 0.05656 at the 24 h), (0.854 µg/mL ± 0.02474, 48 h) and (1.076 µg/mL ± 0.04039, 72 h). In antibacterial assay, the [Ni(L2)] complex presented significant inhibited the bacterial growth of P. aeruginosa (MIC = 256 µg/mL). However, [Ni(L2)] complex did not present clinically relevant minimum inhibitory concentration (MIC ≥1024 µg/mL) against S. aureus and E. coli. The combination of [Ni(L2)] complex and antibacterial drugs resulted in the increased antibiotic activity of gentamicin and amikacin against S. aureus and E.coli multi-resistant strains. Thus, our results showed that [Ni(L2)] complex is a promising molecule for the development of new therapies associated with aminoglycoside antibiotics and in disease control related to resistant bacteria and leishmaniasis.
Asunto(s)
Antibacterianos/farmacología , Complejos de Coordinación/farmacología , Bases de Schiff/farmacología , Tripanocidas/farmacología , Amicacina/farmacología , Animales , Antibacterianos/química , Artemia/efectos de los fármacos , Complejos de Coordinación/química , Sinergismo Farmacológico , Escherichia coli/efectos de los fármacos , Gentamicinas/farmacología , Leishmania infantum/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Níquel/química , Pruebas de Sensibilidad Parasitaria , Pseudomonas aeruginosa/efectos de los fármacos , Bases de Schiff/química , Staphylococcus aureus/efectos de los fármacos , Tripanocidas/químicaRESUMEN
The lectins are carbohydrate-binding proteins that are highly specific to sugar groups associated to other molecules. In addition to interacting with carbohydrates, a number of studies have reported the ability of these proteins to modulate the activity of several antibiotics against multidrug-resistant (MDR) strains. In this study, we report the enhanced antibacterial activity of the gentamicin against MDR strains when complexed with a lectin from Canavalia ensiformis seeds (ConA). Hemagglutination activity test and intrinsic fluorescence spectroscopy revealed that the gentamicin can interact with ConA most likely via the carbohydrate recognition domain (CRD) with binding constant (Kb) value estimated of (0.44 ± 0.04) x 104 M-1. Furthermore, the minimum inhibitory concentrations (MIC) obtained for ConA against all strains studied were not clinically relevant (MIC ≥ 1024 µg/mL). However, when ConA was combined with gentamicin, a significant increase in antibiotic activity was observed against Staphylococcus aureus and Escherichia coli. The present study showed that ConA has an affinity for gentamicin and modulates its activity against MDR strains. These results indicate that ConA improves gentamicin performance and is a promising candidate for structure/function analyses.
Asunto(s)
Canavalia , Gentamicinas , Antibacterianos/farmacología , Gentamicinas/farmacología , Lectinas , Pruebas de Sensibilidad MicrobianaRESUMEN
This study evaluated the effect on enamel demineralization of 10% hydrogen peroxide (H2 O2 ) gels containing different concentrations of sodium trimetaphosphate (TMP) and sodium fluoride (NaF) combined with the daily use of fluoridated or placebo dentifrice. Bovine enamel blocks were selected by surface hardness (n = 72) and randomly assigned to one of the following experimental treatments: 10% H2 O2 ; 10% H2 O2 + 3% TMP + 0.1% NaF; and 10% H2 O2 + 0.3% TMP + 0.05% NaF, each with or without fluoridated dentifrice. H2 O2 -based gels were applied for 30 min d-1 followed by treatment with dentifrice (1 min). Enamels blocks were stored in artificial saliva at 37°C between sessions during the 14 days of experiment. Percentage of surface hardness loss (%SH) was calculated, and the blocks were cut into halves to analyze cross-sectional hardness (ΔKHN). Polarized light microscopy images were obtained of the longitudinal sections of the samples. Enamel treated with fluoridated dentifrice presented lower hardness loss than those treated with placebo dentifrice (%SH and ΔKHN). Use of TMP- and NaF-based gels, regardless of concentration, led to the lowest %SH values. Specimens treated with 10% H2 O2 gel had the highest %SH and ΔKHN values. Gels with 10% H2 O2 + 3% TMP + 0.1% NaF showed the lowest ΔKHN values. Microscopy images clearly showed that the addition of TMP and NaF to the H2 O2 -based gels was effective in reducing the loss of hardness, and the fluoridated dentifrice helped minimize it in all treatments.
Asunto(s)
Blanqueadores , Desmineralización Dental , Animales , Cariostáticos , Bovinos , Estudios Transversales , Esmalte Dental , Fluoruros , Fluoruros Tópicos , Dureza , Peróxido de Hidrógeno , Fluoruro de Sodio , Desmineralización Dental/prevención & controlRESUMEN
Lectins are a group of widely distributed and structurally heterogeneous proteins of nonimmune origin. These proteins have the ability to interact with glycans present on cell surfaces and elicit diverse biological activities. Machaerium acutifolium lectin (MaL) is an N-acetyl-D-glucosamine-binding lectin that exhibits antinociceptive activity via transient receptor potential cation channel subfamily V member 1 (TRPV1). Lectins that have the ability to recognize and interact with N-acetyl-D-glucosamine residues are potential candidates for studies of fungicidal activity. In this work, we show that MaL has antifungal activity against Candida species, and we describe its mode of action towards Candida parapsilosis. MaL inhibited the growth of C. albicans and C. parapsilosis. However, MaL was more potent against C. parapsilosis. The candidacidal mode of action of MaL on C. parapsilosis involves enhanced cell permeabilization, alteration of the plasma membrane proton-pumping ATPase function (H+-ATPase), induction of oxidative stress, and DNA damage. MaL also exhibited antibiofilm activity and noncytotoxicity to Vero cells. These results indicate that MaL is a promising candidate for the future development of a new, natural, and safe drug for the treatment of infections caused by C. parapsilosis.
Asunto(s)
Antifúngicos/farmacología , Candida parapsilosis/metabolismo , Estructuras de la Membrana Celular/química , Fabaceae/química , Lectinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Animales , Antifúngicos/administración & dosificación , Antifúngicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Candida albicans/metabolismo , Candida parapsilosis/citología , Candida parapsilosis/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Estructuras de la Membrana Celular/metabolismo , Chlorocebus aethiops , Medios de Cultivo/análisis , Medios de Cultivo/química , Daño del ADN , Lectinas/administración & dosificación , Lectinas/aislamiento & purificación , Microscopía Electrónica de Rastreo , Propidio/metabolismo , Semillas/química , Células VeroRESUMEN
The use of natural products together with standard antimicrobial drugs has recently received more attention as a strategy to combat infectious diseases caused by multidrug-resistant (MDR) microorganisms. This study aimed to evaluate the capacity of a galactose-binding lectin from Vatairea macrocarpa seeds (VML) to modulate antibiotic activity against standard and MDR Staphylococcus aureus and Escherichia coli bacterial strains. The minimum inhibitory concentration (MIC) obtained for VML against all strains was not clinically relevant (MIC ≥ 1024 µg/mL). However, when VML was combined with the antibacterial drugs gentamicin, norfloxacin and penicillin, a significant increase in antibiotic activity was observed against S. aureus, whereas the combination of VML and norfloxacin presented decreased and, hence, antagonistic antibiotic activity against E. coli. By its inhibition of hemagglutinating activity, gentamicin (MIC = 50 mM) revealed its interaction with the carbohydrate-binding site (CBS) of VML. Using molecular docking, it was found that gentamicin interacts with residues that constitute the CBS of VML with a score of - 120.79 MDS. It is this interaction between the antibiotic and the lectin's CBS that may be responsible for the enhanced activity of gentamicin in S. aureus. Thus, our results suggest that the VML can be an effective modulating agent against S. aureus. This is the first study to report the effect of lectins as modulators of bacterial sensitivity, and as such, the outcome of this study could lay the groundwork for future research involving the use of lectins and conventional antibiotics against such infectious diseases such as community-acquired methicillin-resistant S. aureus (MRSA).
Asunto(s)
Antibacterianos/farmacología , Interacciones Farmacológicas , Fabaceae/química , Galectinas/farmacología , Proteínas de Plantas/farmacología , Staphylococcus aureus/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Semillas/químicaRESUMEN
Gentamicin is an aminoglycoside antibiotic used to treat infections of various origins. In the last few decades, the constant use of gentamicin has resulted in increased bacterial resistance and nephrotoxicity in some cases. In this study, we examined the ability of Dioclea violacea lectin (DVL) in modulate the antimicrobial activity of gentamicin and reduce the nephrotoxicity induced by this drug. The minimum inhibitory concentration (MIC) obtained for DVL against all strains studied was not clinically relevant (MIC ≥ 1024 µg/mL). However, when DVL was combined with gentamicin, a significant increase in antibiotic action was observed against Staphylococcus aureus and Escherichia coli. DVL also reduced antibiotic tolerance in S. aureus during 10 days of continuous treatment. In addition, DVL presented a nephroprotective effect, reducing sodium excretion, N-Gal expression and urinary protein, that are important markers of glomerular and tubular injuries. Taken together, studies of inhibition of hemagglutinating activity, fluorescence spectroscopy and molecular docking revealed that gentamicin can interact with DVL via the carbohydrate recognition domain (CRD), suggesting that the results obtained in this study may be directly related to the interaction of DVL-gentamicin and with the ability of the lectin to interact with glycans present in the cells of the peritoneum.
Asunto(s)
Antibacterianos/farmacología , Dioclea/química , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Gentamicinas/farmacología , Riñón/patología , Lectinas de Plantas/farmacología , Sustancias Protectoras/farmacología , Animales , Antibacterianos/química , Bacterias/efectos de los fármacos , Gentamicinas/química , Hemaglutinación/efectos de los fármacos , Riñón/efectos de los fármacos , Riñón/lesiones , Riñón/fisiopatología , Masculino , Pruebas de Sensibilidad Microbiana , Simulación del Acoplamiento Molecular , Lectinas de Plantas/química , Lectinas de Plantas/aislamiento & purificación , Conejos , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Espectrometría de FluorescenciaRESUMEN
Lectins have been studied in the past few years as an alternative to inhibit the development of pathogenic bacteria and gastrointestinal nematodes of small ruminants. The development of new antibacterial and anthelmintic compounds is necessary owing to the increase in drug resistance among important pathogens. Therefore, this study aimed to evaluate the capacity of a glucose/mannose-binding lectin from Parkia platycephala seeds (PPL) to inhibit the development of Haemonchus contortus and to modulate antibiotic activity against multi-resistant bacterial strains, thereby confirming its efficacy when used in combination with gentamicin. PPL at the concentration of 1.2â¯mg/mL did not show inhibitory activity on H. contortus in the egg hatch test or the exsheathment assay. However, it did show significant inhibition of H. contortus larval development with an IC50 of 0.31â¯mg/mL. The minimum inhibitory concentration (MIC) obtained for PPL against all tested bacterial strains was not clinically relevant (MICâ¯≥â¯1024⯵g/mL). However, when PPL was combined with gentamicin, a significant increase in antibiotic activity was observed against S. aureus and E.coli multi-resistant strains. The inhibition of hemagglutinating activity by gentamicin (MICâ¯=â¯50â¯mM) revealed that it may be interacting with the carbohydrate-binding site of PPL. It is this interaction between the antibiotic and lectin carbohydrate-binding site that may be responsible for the enhanced activity of gentamicin against multi-resistant strains. It can be concluded that PPL showed selective anthelmintic effect, inhibiting the development of H. contortus larvae and that it increased the effect of the antibiotic gentamicin against multi-resistant bacterial strains, thus constituting a potential therapeutic resource against resistant bacterial strains and H. contortus.
Asunto(s)
Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Fabaceae/química , Haemonchus/efectos de los fármacos , Haemonchus/crecimiento & desarrollo , Lectinas/farmacología , Extractos Vegetales/farmacología , Animales , Antihelmínticos/farmacología , Gentamicinas/farmacología , Haemonchus/microbiología , Larva/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Semillas/química , Staphylococcus aureus/efectos de los fármacosRESUMEN
A new mannose/N-acetyl-dglucosamine-specific lectin, named MaL, was purified from seeds of Machaerium acutifolium by precipitation with ammonium sulfate, followed by affinity and ion-exchange chromatography. MaL haemagglutinates either native rabbit erythrocytes or those treated with proteolytic enzymes. MaL is highly stable by the ability to maintain its haemagglutinating activity after exposure to temperatures up to 50⯰C. The lectin haemagglutinating activity was optimum between pH 6.0 and 7.0 and inhibited after incubation with d-mannose and N-acetyl-d-glucosamine and α-methyl-d-mannopyranoside. MaL is a glycoprotein with relative molecular mass of 29â¯kDa (α-chain), 13â¯kDa (ß-chain) and 8â¯kDa (γ-chain) with secondary structure composed of 3% α-helix, 44% ß-sheet, 21% ß-turn, and 32% coil. The orofacial antinociceptive activity of the lectin was also evaluated. MaL (0.03â¯mgâ¯mL-1) reduced orofacial nociception induced by capsaicin, an effect that occurred via carbohydrate recognition domain interaction, suggesting an interaction of MaL with the transient receptor potential cation channel subfamily V member 1 (TRPV1) receptor. Our results confirm the potential pharmacological relevance of MaL as an inhibitor of acute orofacial mediated by TRPV1.
Asunto(s)
Acetilglucosamina/química , Fabaceae/química , Dolor Facial/tratamiento farmacológico , Lectinas/aislamiento & purificación , Lectinas/uso terapéutico , Manosa/química , Canales Catiónicos TRPV/metabolismo , Secuencia de Aminoácidos , Animales , Fenómenos Biofísicos , Cromatografía de Afinidad , Electroforesis en Gel de Poliacrilamida , Femenino , Lectinas/química , Masculino , Estructura Secundaria de Proteína , Conejos , Espectrometría de Masas en Tándem , Pez CebraRESUMEN
Haemonchus contortus is one of the most economically important parasites infecting small ruminants worldwide. This nematode has shown a great ability to develop resistance to anthelmintic drugs, calling for the development of alternative control approaches. Because lectins recognize and bind to specific carbohydrates and glycan structures present in parasites, they can be considered as an alternative to develop new antiparasitic drugs. Accordingly, this work aimed to investigate the anthelmintic effect of Canavalia brasiliensis (ConBr) lectin against H. contortus and to evaluate a possible interaction of ConBr with glycans of this parasite by molecular docking. ConBr showed significant inhibition of H. contortus larval development with an IC50 of 0.26 mg mL-1. Molecular docking assays revealed that glycans containing the core trimannoside [Man(α1-3)Man(α1-6)Man] of H. contortus interact in the carbohydrate recognition domain of ConBr with an interaction value of MDS = -248.77. Our findings suggest that the inhibition of H. contortus larval development is directly related to the recognition of the core trimannoside present in the glycans of these parasites. This work is the first to report on the structure-function relationships of the anthelmintic activity of plant lectins.