ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Nimbolide is one of hundreds of phytochemicals that have been identified within the neem tree (Azadirachta indica A. Juss). As an evergreen tree native to the Indian subcontinent, components of the neem tree have been used for millennia in traditional medicine to treat dental, gastrointestinal, urinary tract, and blood-related ailments, ulcers, headaches, heartburn, and diabetes. In modern times, natural oils and extracts from the neem tree have been found to have activities against a variety of microorganisms, including human pathogens. AIM OF THE STUDY: Helicobacter pylori, a prevalent gastric pathogen, shows increasing levels of antibiotic resistance. Thus, there is an increasing demand for novel therapeutics to treat chronic infections. The in vitro activity of neem oil extract against H. pylori was previously characterized and found to be bactericidal. Given the numerous phytochemicals found in neem oil extract, the present study was designed to define and characterize specific compounds showing bactericidal activity against H. pylori. MATERIALS AND METHODS: Azadirachtin, gedunin, and nimbolide, which are all common in neem extracts, were tested for antimicrobial activity; the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for nine strains of H. pylori. The specific properties of nimbolide were further characterized against H. pylori strain G27. Bactericidal kinetics, reversibility, effectiveness at low pH, and activity under bacteriostatic conditions were examined. The hemolytic activity of nimbolide was also measured. Finally, neem oil extract and nimbolide effectiveness against H. pylori biofilms were examined in comparison to common antibiotics used to treat H. pylori infection. RESULTS: Nimbolide, but not azadirachtin or gedunin, was effective against H. pylori; MICs and MBCs against the nine tested strains ranged between 1.25-5 µg/mL and 2.5-10 µg/mL, respectively. Additionally, neem oil extract and nimbolide were both effective against H. pylori biofilms. Nimbolide exhibited no significant hemolytic activity at biologically relevant concentrations. The bactericidal activity of nimbolide was time- and dose-dependent, independent of active H. pylori growth, and synergistic with low pH. Furthermore, nimbolide-mediated H. pylori cell death was irreversible after exposure to high nimbolide concentrations (80 µg/mL, after 2 h of exposure time and 40 µg/mL after 8 h of exposure). CONCLUSIONS: Nimbolide has significant bactericidal activity against H. pylori, killing both free living bacterial cells as well as cells within a biofilm. Furthermore, the lack of hemolytic activity, synergistic activity at low pH and bactericidal properties even against bacteria in a state of growth arrest are all ideal pharmacological and biologically relevant properties for a potential new agent. This study underscores the potential of neem oil extract or nimbolide to be used as a future treatment for H. pylori infection.
Subject(s)
Azadirachta/chemistry , Helicobacter Infections/drug therapy , Helicobacter pylori/drug effects , Limonins/pharmacology , Anti-Bacterial Agents/pharmacology , Drug Discovery , Glycerides/pharmacology , Humans , Medicine, Traditional/methods , Microbial Sensitivity Tests , Phytochemicals/pharmacology , Plant Extracts/pharmacology , Plant Leaves , Terpenes/pharmacologyABSTRACT
Objective:To find the cytotoxic and apoptotic effects of neem oil extract on the selected cancerous (A-549, PC-3 and DU-145) and noncancerous (NIH3T3 and CCD-18Co) cell lines.Methods:Viability and cytotoxic effect induced by the extract was measured by using MTT assay and apoptotic effect of the extract was evaluated by using Hoechst 33342 and propidium iodide dual staining through a fluorescent microscope and activity of caspases 3, 8 and 9 through colorimetric assay kits.Results:The results showed that neem oil extract significantly reduced the viability in all selected cancer cells treated with varying concentrations of extract as compared with untreated cells and had less effect on noncancerous cell lines. It significantly increased the percentage of necrotic and apoptotic cells, and caspases 3, 8 and 9 activities in all cancer cells treated with extract as compared with untreated cells whereas no effect on noncancerous cell lines. It suggested that neem oil extract exerted a higher cytotoxic effect on cancer cells than normal cells and lower concentration induced apoptosis only in cancer cells. One of the apoptosis-inducing mechanism was through the activation of caspases signaling pathways.Conclusion:Conclusively, it implies that neem oil extract may contain one or more potential agents that can be used as a safe and effective anticancer therapy.
ABSTRACT
Objective: To find the cytotoxic and apoptotic effects of neem oil extract on the selected cancerous (A-549, PC-3 and DU-145) and noncancerous (NIH3T3 and CCD-18Co) cell lines. Methods: Viability and cytotoxic effect induced by the extract was measured by using MTT assay and apoptotic effect of the extract was evaluated by using Hoechst 33342 and propidium iodide dual staining through a fluorescent microscope and activity of caspases 3, 8 and 9 through colorimetric assay kits. Results: The results showed that neem oil extract significantly reduced the viability in all selected cancer cells treated with varying concentrations of extract as compared with untreated cells and had less effect on noncancerous cell lines. It significantly increased the percentage of necrotic and apoptotic cells, and caspases 3, 8 and 9 activities in all cancer cells treated with extract as compared with untreated cells whereas no effect on noncancerous cell lines. It suggested that neem oil extract exerted a higher cytotoxic effect on cancer cells than normal cells and lower concentration induced apoptosis only in cancer cells. One of the apoptosis-inducing mechanism was through the activation of caspases signaling pathways. Conclusion: Conclusively, it implies that neem oil extract may contain one or more potential agents that can be used as a safe and effective anticancer therapy.
ABSTRACT
To evaluate the toxic effect of commercial formulations of neem oil, Azadirachta indica A. Juss, pre-pupae (PP), young pupae (YP) and old pupae (OP) of Diatraea saccharalis F. (Lepidoptera: Crambidae) were sprayed with the diluted extract in distilled water at concentrations of 0.0, 0.3, 0.5, 1.0 and 2.0%. The neem extract caused concentration-dependent effects on mortality of pupae, and the pupae that failed to emerge in adults had multiple abnormalities. The longevity of pupae that emerged in adults (YP and OP group) did not differ from the control group. The abnormalities found in adults were related to mortality in all treatments, except at the concentration of 1.0%. Fertility was assessed according to the oviposition of adult females from the YP group that showed no abnormalities, through the evaluation of the number of deposited eggs and the rate of undeveloped eggs. The results showed a reduction in the number of eggs laid and an increase in the percentage of undeveloped eggs. These results show that neem oil has a high potential to control the toughest stage of the sugarcane borer and also reduces the further development. Therefore, commercial formulations of neem oil have a toxic effect on pupae and adults of D. saccharalis.(AU)
Para avaliar o efeito tóxico de formulações comerciais de óleo de neem, Azadirachta indica A. Juss, pré-pupas (PP), pupas jovens (PJ) e pupas velhas (PV) da Diatraea saccharalis F. (Lepidoptera: Crambidae) foram pulverizadas com o extrato diluído em água destilada, em concentrações de 0,0, 0,3, 0,5, 1,0 e 2,0%. O neem provocou diferentes efeitos sobre a mortalidade de pupas, dependendo da concentração. As pupas que não conseguiram emergir em adultos apresentaram anormalidades múltiplas. Quanto às pupas que emergiram em adultos (grupos PJ e PV), foi calculada a sua longevidade, que não diferiu da do grupo controle. As anormalidades encontradas em adultos estão relacionadas com a mortalidade em todos os tratamentos com exceção da concentração de 1,0%. A fecundidade foi avaliada de acordo com a oviposição de adultos fêmeas do grupo PJ, que não apresentaram anormalidades; dentro dos ovos depositados foi avaliado o número de ovos não desenvolvidos. Os resultados demonstraram redução no número de ovos depositados e aumento na porcentagem de ovos não desenvolvidos. Esses resultados mostraram que o óleo de neem tem elevado potencial para o controle do estágio mais resistente da broca da cana-de-açúcar, além de reduzir o aparecimento das fases subsequentes. Portanto, formulações comerciais de óleo de neem apresentam um efeito tóxico em pupas e adultos de D. Saccharalis.(AU)
Subject(s)
Azadirachta , Saccharum , Toxicity , Insecticides , InsectaABSTRACT
Zearalenone, a mycotoxin produced by fungi of the genus Fusarium, including F. graminearum, triggers reproduction disorders in certain animals and hyperestrogen syndromes in humans. Current research investigates three concentrations of neem oil extract (0.1, 0.25 and 0.5 percent) in reducing the production of zearalenone. Neem oil extract decreased zearalenone amount in the three concentrations but highest inhibition (59.05 percent) occurred at 0.1 percent.
ABSTRACT
Zearalenone, a mycotoxin produced by fungi of the genus Fusarium, including F. graminearum, triggers reproduction disorders in certain animals and hyperestrogen syndromes in humans. Current research investigates three concentrations of neem oil extract (0.1, 0.25 and 0.5%) in reducing the production of zearalenone. Neem oil extract decreased zearalenone amount in the three concentrations but highest inhibition (59.05%) occurred at 0.1%.
