RESUMO
The present study aimed to evaluate the anti-staphylococcal, antibiofilm, cytotoxicity and trypanocidal activity, mechanisms of parasite death and immunomodulatory effect of CrataBL encapsulated into liposomes (CrataBL-Lipo). CrataBL-Lipo were prepared by the freeze-thaw technique and characterized. Anti-staphylococcal and antibiofilm activities of CrataBL and CrataBL-Lipo were evaluated against standard and clinical strains of Staphylococcus aureus susceptible and resistant. Thus, broth microdilution method was performed to determine the Minimum Inhibitory Concentration (MIC). Antibiofilm activity at subinhibitory concentrations was evaluated using the crystal violet staining method. Cytotoxicity of CrataBL-Lipo was verified in L929 fibroblasts and J774A.1 macrophages by determining the inhibitory concentration necessary to kill 50 % of cells (IC50). Trypanocidal activities of CrataBL-Lipo was evaluated in Trypanosoma cruzi and the efficacy was expressed as the concentration necessary to kill 50 % of parasites (EC50). The mechanisms of parasite death and immunomodulatory effect of CrataBL-Lipo were evaluated using flow cytometry analysis. CrataBL-Lipo presented Ø of 101.9 ± 1.3 nm (PDI = 0.245), ζ of +33.8 ± 1.3 mV and %EE = 80 ± 0.84 %. CrataBL-Lipo presented anti-staphylococcal activity (MIC = 0.56 mg/mL to 0.72 mg/mL). CrataBL-Lipo inhibited 45.4 %-75.6 % of biofilm formation. No cytotoxicity of CrataBL-Lipo was found (IC50 > 100 mg/L). CrataBL-Lipo presented EC50 of 1.1 mg/L, presenting autophagy, apoptosis and necrosis as death profile. In addition, CrataBL-Lipo reduced the production of IL-10 and TNF-α levels, causing an immunomodulatory effect. CrataBL-Lipo has a therapeutic potential for the treatment of staphylococcal infections and Chagas disease exhibiting a high degree of selectivity for the microorganism, and immunomodulatory properties.
Assuntos
Antibacterianos , Biofilmes , Lipossomos , Testes de Sensibilidade Microbiana , Staphylococcus aureus , Tripanossomicidas , Trypanosoma cruzi , Biofilmes/efeitos dos fármacos , Trypanosoma cruzi/efeitos dos fármacos , Animais , Camundongos , Staphylococcus aureus/efeitos dos fármacos , Linhagem Celular , Antibacterianos/farmacologia , Tripanossomicidas/farmacologia , Macrófagos/efeitos dos fármacos , Lectinas/farmacologia , Fibroblastos/efeitos dos fármacos , Concentração Inibidora 50 , Sobrevivência Celular/efeitos dos fármacosRESUMO
Punica granatum, popularly known as pomegranate, is a fruit tree with wide worldwide distribution, containing numerous phytochemicals of great medicinal value. The aim of the present study was to determine the phytochemical profile and antioxidant potential of a protein fraction (PF) derived from P. granatum sarcotesta which is rich in lectin. In addition, the acute oral toxicity, genotoxicity and antigenotoxicity of this protein fraction (PF) from P. granatum sarcotesta was measured. The phytochemical profile of PF was determined using HPLC. The in vitro antioxidant effect was assessed using the methods of total antioxidant capacity (TAC) and DPPH and ABTS+ radical scavenging. Acute oral toxicity was determined in female Swiss mice administered a single dose of 2000 mg/kg. This PF was examined for genotoxicity and antigenotoxicity at doses of 500, 1000 and 2000 mg/kg, utilizing mouse peripheral blood cells. Phytochemical characterization detected a high content of ellagic acid and antioxidant capacity similar to that of ascorbic acid (positive control). PF was not toxic (LD50 >2000 mg/kg) and did not exert a genotoxic effect in mice. PF protected the DNA of peripheral blood cells against damage induced by cyclophosphamide. In conclusion, this PF fraction exhibited significant antioxidant activity without initiating toxic or genotoxic responses in mice.
Assuntos
Antioxidantes , Extratos Vegetais , Punica granatum , Animais , Camundongos , Antioxidantes/farmacologia , Feminino , Extratos Vegetais/toxicidade , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Punica granatum/química , Lectinas/toxicidade , Testes de Mutagenicidade , Dano ao DNA/efeitos dos fármacos , Testes de Toxicidade AgudaRESUMO
Wounds encompass physical, chemical, biological, induced damages to the skin or mucous membranes. In wound treatment, combating infections is a critical challenge due to their potential to impede recovery and inflict systemic harm on patients. Previously, the essential oil extracted from Psidium glaziovianum (PgEO) demonstrated antinociceptive and anti-inflammatory attributes, along with negligible oral toxicity. Hence, our study aimed to assess the effects of topically applying a gel formulation containing PgEO to excisional wounds in mice. Additionally, an in vitro antimicrobial assessment was conducted. The formulated gel underwent characterization and toxicological evaluation on erythrocytes, as well as a dermal irritation test. Its antimicrobial activity was tested against both gram-positive and gram-negative bacteria, as well as fungi. Subsequently, an assessment of its efficacy in excisional wound healing was conducted in mice. The findings of this investigation highlight the gel's efficacy against both gram-positive and gram-negative bacteria, as well as fungi. Moreover, this study underscores that the PgEO-gel treatment enhances skin wound healing, potentially due to its capacity to trigger antioxidant enzymes and suppress pro-inflammatory cytokines. Furthermore, the gel exhibited minimal toxicity to erythrocytes and skin irritation. These findings hold promise for prospective preclinical and clinical trials across diverse wound types. In conclusion, this study sheds light on the potential therapeutic applications of the gel formulation containing essential oil from P. glaziovianum in the context of wound healing.
Assuntos
Óleos Voláteis , Psidium , Humanos , Animais , Camundongos , Antibacterianos , Estudos Prospectivos , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Cicatrização , Óleos Voláteis/farmacologiaRESUMO
This study characterized the binding mechanisms of the lectin cMoL (from Moringa oleifera seeds) to carbohydrates using spectroscopy and molecular dynamics (MD). The interaction with carbohydrates was studied by evaluating lectin fluorescence emission after titration with glucose or galactose (2.0-11 mM). The Stern-Volmer constant (Ksv), binding constant (Ka), Gibbs free energy (∆G), and Hill coefficient were calculated. After the urea-induced denaturation of cMoL, evaluations were performed using fluorescence spectroscopy, circular dichroism (CD), and hemagglutinating activity (HA) evaluations. The MD simulations were performed using the Amber 20 package. The decrease in Ksv revealed that cMoL interacts with carbohydrates via a static mechanism. The cMoL bound carbohydrates spontaneously (ΔG < 0) and presented a Ka on the order of 102, with high selectivity for glucose. Protein-ligand complexes were stabilized by hydrogen bonds and hydrophobic interactions. The Hill parameter (h~2) indicated that the binding occurs through the cMoL dimer. The loss of HA at urea concentrations at which the fluorescence and CD spectra indicated protein monomerization confirmed these results. The MD simulations revealed that glucose bound to the large cavity formed between the monomers. In conclusion, the biotechnological application of cMoL lectin requires specific methods or media to improve its dimeric protein structure.
Assuntos
Simulação de Dinâmica Molecular , Moringa oleifera , Ligação Proteica , Sementes , Moringa oleifera/química , Sementes/química , Lectinas de Plantas/química , Multimerização Proteica , Carboidratos/química , Dicroísmo Circular , Lectinas/química , Lectinas/metabolismo , Espectrometria de Fluorescência , Conformação Proteica , Termodinâmica , Ligação de HidrogênioRESUMO
AIMS: We investigated the putative fungistatic and fungicidal activities of pomegranate sarcotesta lectin (PgTeL) against Cryptococcus neoformans B3501 (serotype D), specifically the ability of PgTeL to inhibit yeast capsule and biofilm formation in this strain. METHODS AND RESULTS: PgTeL showed a minimum inhibitory concentration of 172.0 µg ml-1, at which it did not exhibit a fungicidal effect. PgTeL concentrations of 4.0-256.0 µg ml-1 reduced biofilm biomass by 31.0%-64.0%. Furthermore, 32.0-256.0 µg ml-1 PgTeL decreased the metabolic activity of the biofilm by 32.0%-93.0%. Scanning electron microscopy images clearly revealed disruption of the biofilm matrix. Moreover, PgTeL disrupted preformed biofilms. At concentrations of 8.0-256.0 µg ml-1, PgTeL reduced metabolic activity in C. neoformans by 36.0%-92.0%. However, PgTeL did not inhibit the ability of B3501 cells to form capsules under stress conditions. CONCLUSIONS: PgTeL inhibited biofilm formation and disrupted preformed biofilms, demonstrating its potential for use as an anticryptococcal agent.
Assuntos
Criptococose , Cryptococcus neoformans , Punica granatum , Lectinas/farmacologia , Punica granatum/metabolismo , Plâncton/metabolismo , Biofilmes , Testes de Sensibilidade Microbiana , Antifúngicos/farmacologia , Antifúngicos/metabolismoRESUMO
Melanoma is difficult to treat with chemotherapy, prompting the need for new treatments. Protease inhibitors have emerged as promising candidates as tumor cell proteases promote metastasis. Researchers have developed a chimeric form of the Bauhinia bauhinioides kallikrein inhibitor, rBbKIm, which has shown negative effects on prostate tumor cell lines DU145 and PC3. Crataeva tapia bark lectin, CrataBL, targets sulfated oligosaccharides in glycosylated proteins and has also demonstrated deleterious effects on prostate and glioblastoma tumor cells. However, neither rBbKIm nor its derived peptides affected the viability of SK-MEL-28, a melanoma cell line, while CrataBL decreased viability by over 60%. Two peptides, Pep. 26 (Ac-Q-N-S-S-L-K-V-V-P-L-NH2) and Pep. 27 (Ac-L-P-V-V-K-L-S-S-N-Q-NH2), were also tested. Pep. 27 suppressed cell migration and induced apoptosis when combined with vemurafenib, while Pep. 26 inhibited cell migration and reduced nitric oxide and the number of viable cells. Vemurafenib, a chemotherapy drug used to treat melanoma, was found to decrease the release of interleukin 8 and PDGF-AB/BB cytokines and potentiated the effects of proteins and peptides in reducing these cytokines. These findings suggest that protease inhibitors may be effective in blocking melanoma cells and highlight the potential of CrataBL and its derived peptides.
Assuntos
Melanoma , Masculino , Humanos , Vemurafenib/farmacologia , Melanoma/tratamento farmacológico , Linhagem Celular Tumoral , Apoptose , Citocinas/farmacologia , Inibidores de Proteases/farmacologiaRESUMO
The ethanolic extract from Croton blanchetianus leaves has been shown to have antinociceptive activity in mice. Here, we investigated the antinociceptive activity of an ethyl acetate fraction (EAF) from this extract in mice and the possible pathways involved in the analgesic effect. Adverse effects on behavior and motor coordination were also evaluated. The EAF was characterized by liquid chromatography coupled with mass spectrometry and evaluated (12.5, 25, and 50â mg/kg per os) in the acetic acid-induced abdominal writhing, formalin, hot plate, and tail immersion assays. Naloxone, atropine, glibenclamide, prazosin, or yohimbine was pre-administered to mice to investigate the involved pathways in the formalin test. The open-field, rotarod, and elevated plus-maze tests were used to assess behavior and locomotion. The main components of the EAF were quercetin-3-O-(2-rhamnosyl) rutinoside, hyperoside, quercetin rutinoside pentoside, and quercetin hexoside deoxyhexoside. EAF showed antinociceptive effects in all models and was effective against both neurogenic and inflammatory pain. The reversion of the effects in the formalin test by naloxone and atropine revealed that the EAF acted via the opioid and cholinergic systems. In the open-field test, the behavior of the animals treated with the EAF was like that of control, except at the highest dose, when hypnosis, eyelid ptosis, decreased walking, hygiene, and rearing behaviors were observed. No muscle relaxant effect was observed, but an anxiogenic effect was observed at all doses. This study provides new scientific evidence on the pharmacological properties of C.â blanchetianus leaves and their potential for the development of phytomedicines with analgesic properties.
Assuntos
Croton , Euphorbiaceae , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Analgésicos Opioides/farmacologia , Animais , Colinérgicos , Flavonoides , Camundongos , Extratos Vegetais/uso terapêutico , Folhas de PlantaRESUMO
Lippia alba is an aromatic shrub known to produce a diversity of essential oils, which can be classified into chemotypes. This study reports on the insecticidal activity of essential oil from L. alba leaves collected at Caatinga and its major compound against termite Nasutitermes corniger and maize weevil Sitophilus zeamais. The chromatographic analysis revealed the presence of 19 compounds, with 1,8-cineole being the most common (70.01%). When ingested, the oil promoted the mortality of N. corniger (LC50: 18.25 and 8.4 nL/g for workers and soldiers, respectively). The compound 1,8-cineole was also termiticidal for workers (LC50: 13.7 nL/g). The oil inhibited the activity of N. corniger exoglucanase, xylanase, and proteases. Toxicity by ingestion to S. zeamais was detected for the oil (LC50: 0.297 µL/g) but not for 1,8-cineole; however, both the oil and 1,8-cineole showed anti-nutritional effects. Fumigant effects of the oil and 1,8-cineole against S. zeamais (LC50 of 78.0 and 13.64 µL/L in air, respectively) were detected. This is the first record of a chemotype VI oil from L. alba collected at Caatinga and the first report of the insecticidal activity of a chemotype VI oil. Our study demonstrates that essential oil from L. alba and 1,8 cineole have the potential for the development of natural insecticides.
Assuntos
Inseticidas , Isópteros , Lippia , Óleos Voláteis , Gorgulhos , Animais , Eucaliptol , Inseticidas/farmacologia , Óleos Voláteis/farmacologia , Folhas de PlantaRESUMO
Previous studies have characterized a saline extract from Microgramma vacciniifolia rhizome and its lectin (MvRL)-rich fraction with low acute toxicity. In the present study, we evaluated these preparations for acute toxicity (1,000â mg/kg) and antinociceptive and anti-inflammatory activities (100-400â mg/kg for the extract and 25-50â mg/kg for the fraction). No signs of toxicity were observed. Both the extract and fraction increased the latency period for nociception in the hot plate assay, decreased writhing induced by acetic acid, and promoted analgesic effects in phases 1 and 2 of the formalin test. The antinociceptive mechanism was attributed to interactions with opioid receptors and K+ ATPase channels. The extract and fraction decreased carrageenan-induced paw edema in 46.15 % and 77.22 %, respectively, at the highest doses evaluated. Furthermore, the fraction was shown to act on the bradykinin pathway. The ability to decrease leukocyte migration after treatment was also verified in the peritonitis and air pouch models. In exudates collected from air pouches, decreased tumor necrosis factor (TNF)-α and increased interleukin (IL)-10 levels were noted. Both the extract and fraction also effectively inhibited the development of granulomatous tissue. In conclusion, the substances investigated in this study can be used for the development of novel therapeutic options for pain and inflammatory processes.
Assuntos
Analgésicos/farmacologia , Anti-Inflamatórios/farmacologia , Edema/tratamento farmacológico , Lectinas/química , Extratos Vegetais/farmacologia , Rizoma/química , Analgésicos/química , Analgésicos/isolamento & purificação , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Carragenina , Movimento Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Edema/induzido quimicamente , Leucócitos/efeitos dos fármacos , Camundongos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Receptores Opioides/metabolismo , Salinidade , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
The glutathione S-transferases (GSTs) are enzymes involved in several distinct biological processes. In insects, the GSTs, especially delta and epsilon classes, play a key role in the metabolism of xenobiotics used to control insect populations. Here, we investigated its potential role in temephos resistance, examining the GSTE2 gene from susceptible (RecL) and resistant (RecR) strains of the mosquito Aedes aegypti, vector for several pathogenic arboviruses. Total GST enzymatic activity and the GSTE2 gene expression profile were evaluated, with the GSTE2 cDNA and genomic loci sequenced from both strains. Recombinant GSTE2 and mutants were produced in a heterologous expression system and assayed for enzyme kinetic parameters. These proteins also had their 3D structure predicted through molecular modeling. Our results showed that RecR has a profile of total GST enzymatic activity higher than RecL, with the expression of the GSTE2 gene in resistant larvae increasing six folds. Four exclusive RecR mutations were observed (L111S, I150V, E178A and A198E), which were absent in the laboratory susceptible strains. The enzymatic activity of the recombinant GSTE2 showed different kinetic parameters, with the GSTE2 RecR showing an enhanced ability to metabolize its substrate. The I150V mutation was shown to induce significant changes in catalytic parameters and a 3D modeling of GSTE2 mapped two of the RecR changes (L111S and I150V) near the enzyme's catalytic pocket, also implying an impact on its catalytic activity. Our results reinforce a potential role for GSTE2 in the metabolic resistance phenotype while contributing to the understanding of the molecular basis for the resistance mechanism.
Assuntos
Aedes , Inseticidas , Animais , Resistência a Inseticidas , Mosquitos Vetores , TemefósRESUMO
Our objectives were to identify genes of the multi-drug efflux system and to evaluate the antimicrobial activities of polypyrrole nanoparticles (PPy-NPs) and aqueous extract of Moringa oleifera against Staphylococcus spp. isolated from dairy farms in Northeast Brazil. Initially, 162 Staphylococcus spp. isolates were subjected to in vitro antimicrobial sensitivity tests. Of these, 35 presented antimicrobial multi-drug resistance phenotypes. These 35 isolates were then referred for the detection of norA, norB, norC, msrA, mgrA, tet-38, and lmrS genes, all of which feature in multi-drug efflux systems. In the isolates carrying the genes, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs and Moringa oleifera aqueous extract were determined. In the molecular analysis of the 35 isolates norA, norC, tet-38, and msrA genes were detected and for the other genes norB, lmrS and mgrA there was no amplification. Antimicrobial activity was verified of PPy-NPs and aqueous extract of Moringa oleifera in Staphylococcus spp. carrying multi-drug efflux system genes. We concluded that there are multi-drug efflux system genes present in the Staphylococcus spp. from the agricultural environment in Northeast Brazil, and that aqueous extract of Moringa oleifera and PPy-NPs show bactericidal activity against these isolates.
Assuntos
Antibacterianos/farmacologia , Moringa oleifera/química , Nanopartículas/química , Extratos Vegetais/farmacologia , Polímeros/química , Pirróis/química , Staphylococcus/efeitos dos fármacos , Animais , Antibacterianos/química , Bovinos , Indústria de Laticínios , Farmacorresistência Bacteriana Múltipla , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Extratos Vegetais/química , Staphylococcus/genéticaRESUMO
Aedes aegypti control is a key component of the prophylaxis of dengue fever and other diseases. Moringa oleifera seeds contain a water-soluble lectin (WSMoL) with larvicidal and ovicidal activities against this insect. In this study, A. aegypti individuals were exposed at the third larval instar for 24â¯h to the water extract (0.1-1.0â¯mg/mL of protein) or lectin-rich fraction (0.05-0.6â¯mg/mL of protein) containing WSMoL, and then their survival and development were followed for 9 days post-exposure. The feeding capacity of adult females that developed from the treated larvae and the hatching success of eggs laid by them were also evaluated. Further, any alterations to the midgut histology of treated larvae, pupae, and adults were investigated. The extract and fraction induced the death of A. aegypti larvae along the post-exposure period. Both preparations also delayed the developmental cycle. The midguts of treated larvae and pupae showed disorganization and epithelial vacuolization, while in treated adults, the epithelium was underdeveloped compared to control. Unlike in control mosquitos, proliferating cells were not detected in treated larvae, and appeared in lower numbers in treated pupae than in control pupae. Adult females that developed from larvae treated with the fraction gained less weight after a blood meal compared with control. The amount of eggs laid by females that developed from larvae treated with both the extract and fraction was significantly lower than in control. In addition, the eggs showed lower hatching rates. In conclusion, females that developed from larvae treated with both the water extract and lectin-rich fraction showed reduced engorgement after a blood meal, with the consequent impairment of their fertility and fecundity. These results were probably due to the damage to midgut organization and impairment of the remodeling process during metamorphosis.
Assuntos
Aedes/efeitos dos fármacos , Lectinas/farmacologia , Moringa oleifera/química , Extratos Vegetais/farmacologia , Aedes/crescimento & desenvolvimento , Aedes/fisiologia , Animais , Feminino , Fertilidade/efeitos dos fármacos , Inseticidas/química , Inseticidas/farmacologia , Intestinos/efeitos dos fármacos , Intestinos/crescimento & desenvolvimento , Intestinos/patologia , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lectinas/química , Controle de Mosquitos , Extratos Vegetais/química , Pupa/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Pupa/fisiologia , Sementes/química , Água/químicaRESUMO
Currently available drugs for treatment of glioblastoma, the most aggressive brain tumor, remain inefficient, thus a plethora of natural compounds have already been shown to have antimalignant effects. However, these have not been tested for their impact on tumor cells in their microenvironment-simulated cell models, e.g., mesenchymal stem cells in coculture with glioblastoma cell U87 (GB). Mesenchymal stem cells (MSC) chemotactically infiltrate the glioblastoma microenvironment. Our previous studies have shown that bone-marrow derived MSCs impair U87 growth and invasion via paracrine and cell-cell contact-mediated cross-talk. Here, we report on a plant-derived protein, obtained from Crataeva tapia tree Bark Lectin (CrataBL), having protease inhibitory/lectin activities, and demonstrate its effects on glioblastoma cells U87 alone and their cocultures with MSCs. CrataBL inhibited U87 cell invasion and adhesion. Using a simplified model of the stromal microenvironment, i.e., GB/MSC direct cocultures, we demonstrated that CrataBL, when added in increased concentrations, caused cell cycle arrest and decreased cocultured cells' viability and proliferation, but not invasion. The cocultured cells' phenotypes were affected by CrataBL via a variety of secreted immunomodulatory cytokines, i.e., G-CSF, GM-CSF, IL-6, IL-8, and VEGF. We hypothesize that CrataBL plays a role by boosting the modulatory effects of MSCs on these glioblastoma cell lines and thus the effects of this and other natural lectins and/or inhibitors would certainly be different in the tumor microenvironment compared to tumor cells alone. We have provided clear evidence that it makes much more sense testing these potential therapeutic adjuvants in cocultures, mimicking heterogeneous tumor-stroma interactions with cancer cells in vivo. As such, CrataBL is suggested as a new candidate to approach adjuvant treatment of this deadly tumor.
Assuntos
Capparaceae/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Casca de Planta/química , Extratos Vegetais/farmacologia , Lectinas de Plantas/farmacologia , Inibidores de Proteases/farmacologia , Adesão Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Citocinas/biossíntese , Glioblastoma/metabolismo , Humanos , Células-Tronco Mesenquimais/metabolismo , Metaloproteases/antagonistas & inibidores , Óxido Nítrico/biossíntese , Extratos Vegetais/química , Lectinas de Plantas/química , Inibidores de Proteases/químicaRESUMO
The Alpinia purpurata inflorescence contains a lectin (ApuL), which has immunomodulatory activities on human cells. In the present work, it was evaluated the antibacterial and antifungal effects of ApuL against human pathogens. ApuL showed bacteriostatic activity against non-resistant (UFPEDA-02) and an oxacillin-resistant isolate (UFPEDA-672) of Staphylococcus aureus with minimal inhibitory concentrations (MIC50) of 50 and 400⯵g/mL, respectively. In addition, it showed bactericidal effect on the non-resistant isolate (minimal bactericidal concentration: 200⯵g/mL). For Candida albicans and Candida parapsilosis, ApuL showed fungistatic effect (MIC50: 200 and 400⯵g/mL, respectively). The lectin was able to impair the viability of the microorganism cells, as indicated by propidium iodide (PI) staining. Analysis of growth curves, protein leakage, and ultrastructural changes supported that ApuL acts through distinct mechanisms on S. aureus isolates. Ultrastructural analysis of ApuL-treated Candida cells revealed malformations with elongations and bulges. ApuL-oxacillin combination showed synergistic effect on the oxacillin-resistant isolates UFPEDA-670 and 671, which were not sensitive to lectin alone. Synergism was also detected for ApuL-ceftazidime against a multidrug-resistant isolate of Pseudomonas aeruginosa. Synergistic action of ApuL-fluconazole was detected for C. parapsilosis, which was insensitive to the drug alone. Biofilm formation by S. aureus non-resistant isolate and C. albicans was remarkably inhibited by ApuL at sub-inhibitory concentrations. In conclusion, ApuL showed differential effects on non-resistant and resistant bacterial isolates, was active against Candida species, and showed synergistic action in combination with antibiotics.
Assuntos
Alpinia/química , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Lectinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Candida albicans/crescimento & desenvolvimento , Candida albicans/fisiologia , Sinergismo Farmacológico , Lectinas/isolamento & purificação , Testes de Sensibilidade Microbiana , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/fisiologiaRESUMO
People broadly use medicinal plants although their safety and efficacy are often neglected. Several Cactaceae plants are used for medicinal purposes, but Pilosocereus gounellei (xique-xique) has been little studied for their pharmacological properties. In this study, a saline extract from Pilosocereus gounellei stem was evaluated for acute toxicity, effects on motor function, and antinociceptive activity in mice. Thin-layer chromatography and high-performance liquid chromatography revealed the presence of flavonoids and reducing sugars. No death of mice was noted with oral treatments at 2000 and 5000â¯mg/kg. Body weight gain, food intake, and water consumption were not affected with treatment at 2000â¯mg/kg. The number of segmented leukocytes was higher in the treated mice than in control, and reduced serum levels of cholesterol and triglycerides were detected after both extract treatments. No histological alterations in the liver, heart, lung, kidney, and spleen were detected. Open field and rotarod tests showed that the extract did not affect motor function and exploratory activity. Antinociceptive activity was detected in acetic acid-induced writhing, tail-immersion, and formalin tests, indicating interference with both non-inflammatory and inflammatory pain. In conclusion, the P. gounellei extract has antinociceptive activity in mice without showing toxicity and altering motor coordination.
Assuntos
Analgésicos/uso terapêutico , Cactaceae , Dor/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Ácido Acético , Analgésicos/toxicidade , Animais , Artemia/efeitos dos fármacos , Colesterol/sangue , Feminino , Formaldeído , Temperatura Alta/efeitos adversos , Masculino , Camundongos , Dor/etiologia , Extratos Vegetais/toxicidade , Caules de Planta/química , Desempenho Psicomotor/efeitos dos fármacos , Cloreto de Sódio/química , Triglicerídeos/sangueRESUMO
BACKGROUND: Spondias tuberosa is a plant that produces a fruit crop with high economic relevance at Brazilian Caatinga. Its roots and leaves are used in folk medicine. METHODS: Chemical composition of a hexane extract from S. tuberosa leaves was evaluated by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and 1H nuclear magnetic resonance (NMR). Antioxidant potential was investigated by DPPH and ABTS assays. Antifungal action on Candida species was evaluated determining the minimal inhibitory concentration (MIC50) and putative mechanisms were determined by flow cytometry analysis. In addition, hemolytic activity on human erythrocytes was assessed and the concentration required to promote 50% hemolysis (EC50) was determined. RESULTS: Phytochemical analysis by TLC showed the presence of flavonoids, hydrolysable tannins, saponins and terpenes. The HPLC profile of the extract suggested the presence of gallic acid (0.28 ± 0.01 g%) and hyperoside (1.27 ± 0.01 g%). The representative 1H NMR spectrum showed saturated and unsaturated fatty acids among the main components. The extract showed weak and moderate antioxidant activity in DPPH (IC50: 234.00 µg/mL) and ABTS (IC50: 123.33 µg/mL) assays, respectively. It was able to inhibit the growth of C. albicans and C. glabrata with MIC50 of 2.0 and 0.078 mg/mL, respectively. The treatment of C. glabrata cells with the extract increased levels of mitochondrial superoxide anion, caused hyperpolarization of mitochondrial membrane, and compromised the lysosomal membrane. Weak hemolytic activity (EC50: 740.8 µg/mL) was detected. CONCLUSION: The results demonstrate the pharmacological potential of the extract as antioxidant and antifungal agent, aggregating biotechnological value to this plant and stimulating its conservation.
Assuntos
Anacardiaceae/química , Antifúngicos/farmacologia , Antioxidantes/farmacologia , Membranas Mitocondriais/efeitos dos fármacos , Extratos Vegetais/farmacologia , Candida/citologia , Candida/efeitos dos fármacos , Hexanos , Lisossomos/efeitos dos fármacosRESUMO
Biotechnological techniques allow the investigation of alternatives to outdated chemical insecticides for crop protection; some investigations have focused on the identification of molecules tailored from nature for this purpose. We, herein, describe the negative effects of water-soluble lectin from Moringa oleifera seeds (WSMoL) on Anagasta kuehniella development. The chitin-binding lectin, WSMoL, impaired the larval weight gain by 50% and affected the activity of the pest's major digestive enzymes. The commitment of the digestive process became evident after controlled digestion studies, where the capacity of protein digestion was compromised by >90%. Upon acute exposure, the lectin was not resistant to digestion; however, chronic ingestion of WSMoL was able to reverse this feature. Thus, we show that resistance to digestion may not be a prerequisite for a lectin's ability to exert negative effects on larval physiology. The mechanism of action of WSMoL involves binding to chitin with possible disruption to the peritrophic membrane, causing disorder between the endo- and ectoperitrophic spaces. Additionally, results suggest that WSMoL may trigger apoptosis in gut cells, leading to the lower enzymatic activity observed in WSMoL-fed larvae. Although assays employing an artificial diet did not demonstrate effects of WSMoL on A. kuehniella mortality, this lectin may hold potential for exerting insecticide effects on other pest insects, as well for use in other experimental approaches, such as WSMoL-expressing plants. Moreover, the use of WSMoL with other biotechnological tools, such as 'pyramid' crops, may represent a strategy for delaying the evolution of pest resistance to transgenic crops, since its multiple site targets could act in synergism with other insecticide compounds.
Assuntos
Inseticidas/farmacologia , Lectinas/farmacologia , Moringa oleifera/química , Mariposas/fisiologia , Animais , Quitina/química , Fenômenos Fisiológicos do Sistema Digestório/efeitos dos fármacos , Inseticidas/química , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lectinas/química , Mariposas/efeitos dos fármacos , Mariposas/crescimento & desenvolvimento , Sementes/químicaRESUMO
Essential oil from fresh leaves of the shrub Piper corcovadensis (Miq.) C. DC was obtained in 0.21% (w/w) yield by hydrodistillation in a Clevenger type apparatus. Thirty-one components, accounting for 96.61% of the leaf oil, were identified by gas chromatography-mass spectrometry. The major constituents of the oil were 1-butyl-3,4-methylenedioxybenzene (30.62%), terpinolene (17.44%), trans -caryophyllene (6.27%), α-pinene (5.92%), δ-cadinene (4.92%), and Limonene (4.46%). Bioassays against larvae of the Dengue mosquito (Aedes aegypti) revealed that leaf oil (LC50 = 30.52 ppm), terpinolene (LC50 = 31.16 ppm), and pure 1-butyl-3,4-methylenedioxybenzene (LC50 = 22.1 ppm) possessed larvicidal activities and are able to interfere with the activity of proteases from L4 gut enzymes. Additionally, the essential oil exhibited a strong oviposition deterrent activity at 50 and 5 ppm. This paper constitutes the first report of biological activities associated with the essential oil of leaves of P. corcovadensis.
Assuntos
Aedes , Mosquitos Vetores , Óleos Voláteis/química , Piper/química , Aedes/efeitos dos fármacos , Aedes/fisiologia , Animais , Monoterpenos Bicíclicos , Bioensaio , Monoterpenos Cicloexânicos , Cicloexenos/análise , Dengue/transmissão , Eletroforese em Gel de Poliacrilamida , Cromatografia Gasosa-Espectrometria de Massas , Larva/enzimologia , Limoneno , Monoterpenos/análise , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/fisiologia , Óleos Voláteis/isolamento & purificação , Óleos Voláteis/farmacologia , Oviposição/efeitos dos fármacos , Peptídeo Hidrolases/análise , Folhas de Planta/química , Sesquiterpenos Policíclicos , Inibidores de Proteases/farmacologia , Sesquiterpenos/análise , Terpenos/análise , Tripsina/efeitos dos fármacos , Tripsina/metabolismoRESUMO
Moringa oleifera flower extract, with trypsin inhibitor activity, is a larvicidal agent on Aedes aegypti. This work reports the isolation of trypsin inhibitor (M. oleifera flower trypsin inhibitor (MoFTI)) and its effect on A. aegypti egg hatching, viability of newly hatched larvae, survival of pupae, and growth of inhabitant bacteria from midgut of fourth-instar larvae (L4). MoFTI (K i, 2.4 µM), isolated by affinity chromatography on trypsin-agarose column, was an 18.2 kDa polypeptide on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Flower extract (at concentrations of 8.5-17.0 mg/mL) reduced egg hatchability while MoFTI (0.05-0.5 mg/mL) did not affect the hatching rate. Mortality of newly hatched larvae ranged from 3.5 to 19.1 % in the presence of the extract (4.0-17.0 mg/mL) and was also promoted by MoFTI (LC50, 0.3 mg/mL). After 72 h, larvae incubated with extract at 13.0 and 17.0 mg/mL were at stages L2 and L1, respectively, while in control they reached L3 instar. In the presence of MoFTI, at all concentrations tested, the larvae did not pass the first instar. Flower extract and MoFTI did not interfere on pupae survival. The extract and MoFTI inhibited the growth of L4 gut bacteria (minimum inhibitory concentrations of 3.47 and 0.031 mg/mL, respectively) but only the inhibitor showed bactericide effect (minimum bactericidal concentration of 1.0 mg/mL). The findings reported herein indicate that MoFTI constitutes a larvicidal principle from M. oleifera flowers against A. aegypti newly hatched larvae and is an antibacterial agent active against the microbiota from L4 gut.
Assuntos
Aedes/crescimento & desenvolvimento , Aedes/microbiologia , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Inseticidas/farmacologia , Moringa oleifera/química , Inibidores da Tripsina/farmacologia , Aedes/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Sistema Digestório/microbiologia , Flores/química , Inseticidas/análise , Inseticidas/isolamento & purificação , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/microbiologia , Óvulo/efeitos dos fármacos , Óvulo/fisiologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Pupa/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Tripsina/metabolismo , Inibidores da Tripsina/isolamento & purificaçãoRESUMO
The water-soluble lectin isolated from Moringa oleifera seeds (WSMoL) is a larvicidal, ovicidal, and oviposition-stimulating agent against Aedes aegypti under laboratory conditions. This study investigated the effect of WSMoL in traps for the capture of A. aegypti eggs and adult females under semi-field conditions and determined whether gravid females could detect WSMoL by an olfactory response. WSMoL was isolated according to a previously described procedure using chitin chromatography. The bioassays were performed in large cages (12.5 m(3)). Two traps for collection of eggs (ovitrap) or adult mosquitoes (MosquiTRAP(TM)) were placed in a cage. One was filled with WSMoL (0.1 mg/mL) and the other with tap water (negative control). An infusion of Panicum maximum leaves was used as a positive control. Forty gravid females were then released in each cage. After 2 (for oviposition) or 3 h (for female capture), the traps were removed, and the number of eggs or females was counted. An olfactometry assay was performed to investigate whether the effect of WSMoL on gravid females was linked to an olfactory response. WSMoL showed an oviposition-stimulating effect (65 ± 14%) that was similar (p < 0.05) to that promoted by the P. maximum infusion (67 ± 11%). The efficiency of MosquiTRAP(TM) in capturing gravid females was not increased by WSMoL. The olfactometry assay indicated that the response of females to WSMoL did not involve the stimulation of olfactory sensilla. WSMoL effectively captured eggs when used in ovitraps under semi-field conditions; this property, together with the ovicidal and larvicidal activities of this lectin, makes it an interesting candidate for A. aegypti control.