RESUMO
Tuberculosis (TB) is a life threatening infectious disease which is prevalent throughout the world. Mycobacterium bovis based Bacille Calmette-Gue´rin (BCG) is the only vaccine available against TB however, this (single) vaccine is not enough to eradicate it. Furthermore, numbers of researches from different parts of the World have shown its efficacy as variable. Hence other (better) vaccines like DNA vaccines are needed in addition to BCG in order to achieve desired goal of TB eradication. The current study was aimed to develop subunit based DNA vaccines against TB and to check their efficacy. Two constructs Bfrb-pND14 and Mpt32-pND14 were made and used as DNA vaccines. Endotoxin free DNA preparations were made and used in immunization studies. Twenty Balb/c female mice of age eight weeks were used in trial. Two experimental groups each comprising eight animals were used to inoculate Mpt32-pND14 and Bfrb-pND14 vaccines respectively. A group of four animals was used as negative control. Animals were bled through tail periodically and finally through cardiac puncture before euthanization. Antibodies were confirmed through dot blot and Agar Gel Immuno Diffusion test (AGID). All the animals immunized with both vaccines were found positive as tested through dot blot and AGID. The results of this study have indicated that both the M. tb genes have produced strong immune response in mice model through pND14 vector and proved themselves as good subunit based DNA vaccines.
Assuntos
Proteínas de Bactérias/administração & dosagem , Ferritinas/administração & dosagem , Mycobacterium tuberculosis/genética , Vacinas contra a Tuberculose/administração & dosagem , Tuberculose/prevenção & controle , Vacinas de DNA/administração & dosagem , Animais , Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Feminino , Ferritinas/genética , Ferritinas/imunologia , Imunização , Imunogenicidade da Vacina , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/imunologia , Tuberculose/imunologia , Tuberculose/microbiologia , Vacinas contra a Tuberculose/imunologia , Vacinas de DNA/imunologiaRESUMO
Background The aim of this experiment was to evaluate the cytotoxic, thrombolytic, analgesic, sedative-hypnotic and anxiolytic activities of the methanolic extract of Ficus cunia leaves. Methods Primary phytochemical screening was accomplished by using established methods. Cytotoxicity was studied by brine shrimp lethality test, and the thrombolytic assay was conducted through clot lysis method with human blood. The in vivo action was done using mice of both sexes. The analgesic activity was evaluated by acetic acid-induced writhing test and formalin-induced paw licking test. Open field, hole cross and thiopental Na-induced sleeping time test were used to examine the sedative-hypnotic activity, and elevated plus maze (EPM) and hole board test were used to identify the anxiolytic activity. Results The results elicited that the extract contained several phytochemicals such as alkaloid, flavonoid, and tannin. The extract was found to have a median lethal concentration (LC50) value of 55.48 µg/mL in the brine shrimp lethality bioassay. It was also assessed for antithrombotic activity when compared with streptokinase; it has significant (p < 0.001) thrombolytic effect (34.72 ± 1.74%) contrasted with standard streptokinase (67 ± 1.56%). The extract at doses of 200 and 400 mg/kg produced inhibition of 32.58% and 46.63% in acetic acid-induced pain and 45.88 and 61.18% in formalin-induced pain. The sedative and hypnotic activities on the central nervous system of the methanol extract of F. cunia (MEFC) leaves were evaluated. The extract delivered critical sedative impact at the doses of 200 and 400 mg/kg (by oral route) treated with reference to the substance diazepam, and the hypnotic impact was also observed in the case of mice. MEFC at its maximum dose (400 mg/kg) significantly (p < 0.01) increased the time spent in the open arms of the EPM. In the hole board test, there was a dose-dependent (at 200 and 400 mg/kg) and a significant (p < 0.05 and p < 0.01) increase in the number of head pokes in comparison to control. Conclusions The results of the present study gave a helpful baseline in progression for the possible use of MEFC as a cytotoxic, thrombolytic, analgesic, sedative-hypnotic and anxiolytic drug.
Assuntos
Analgésicos/farmacologia , Fibrinolíticos/farmacologia , Ficus/química , Metanol/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Animais , Artemia/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Feminino , Humanos , Hipnóticos e Sedativos/farmacologia , Camundongos , Dor/tratamento farmacológico , Fitoterapia/métodosRESUMO
Background: The present study was conducted to investigate the antinociceptive activity of methanol extract of Macaranga denticulata (Met.MD) in an animal model, followed by molecular docking analysis. Methods: Antinociceptive activity was determined by acetic acid-induced writhing and formalin-induced licking test in mice. Then, molecular docking study was performed to identify compounds having maximum activity against the COX-1 enzyme using Schrödinger Maestro (version 10.1) to determine docking fitness. Results: A preliminary phytochemical analysis of Met.MD revealed that it contained alkaloids, carbohydrates, phenols, flavonoids, tannins, and terpenoids. Met.MD exhibited a dose-dependent and statistically significant antinociceptive activity in the acetic acid and formalin test at the doses of 200 and 400 mg/kg. In addition, our docking study showed that macarangin had the best fitness score of -5.81 with COX-1 enzyme among six major compounds of M. denticulata. Conclusions: Results of the present study confirmed the potential antinociceptive activity of M. denticulata leaf extract in both in vivo and in silico models.