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Cultivo de células en suspensión de Azadirachta indica A. Juss para la producción de metabolitos secundarios con actividad antiplasmodial
dc.rights.license | Atribución-NoComercial 4.0 Internacional |
dc.contributor.advisor | Orozco-Sánchez, Fernando |
dc.contributor.author | Zuleta-Castro, Leidy Carolina |
dc.date.accessioned | 2020-04-27T20:55:39Z |
dc.date.available | 2020-04-27T20:55:39Z |
dc.date.issued | 2020-04-23 |
dc.identifier.uri | https://repositorio.unal.edu.co/handle/unal/77450 |
dc.description.abstract | La malaria causa alrededor de 500.000 muertes anuales en el mundo. El parásito causante de la enfermedad (Plasmodium sp) ha adquirido resistencia a los medicamentos actuales lo que obliga a la continua búsqueda de nuevas sustancias bioactivas para su control. En esta investigación, se establecieron condiciones de cultivo para células de Azadirachta indica que favorecieran la producción de metabolitos secundarios con actividad antiplasmodial in vitro sobre la cepa FCR3 de Plasmodium falciparum. A partir de la biomasa cultivada bajo tales condiciones, se obtuvieron extractos y con ellos se realizaron ensayos de actividad sobre el parásito. Adicionalmente, se llevó a cabo un estudio de actividad biológica para determinar el estadío específico sobre el cual actuaba el extracto más activo y posteriormente, este mismo fue analizado mediante UPLC-ESI-Qtof-MS con el fin de identificar tentativamente algunos compuestos que pudiesen estar implicados en la actividad antiplasmodial. Las mejores combinaciones a nivel de matraz Erlenmeyer fueron: 25°C, 40 gSac/L, sin adición de peróxido de hidrógeno y empleando matraces Erlenmeyer sin bafles (IC50 = 26.16 µg/mL) y 15 °C, empleando medio MS, en presencia del precursor acetato de sodio, sin adición de elicitores y 30 gSac/L) (IC50 = 18.72 µg/mL, mejor actividad a escala de matraz). Sin embargo, la actividad antiplasmodial in vitro fue principalmente promovida en biorreactor de tanque agitado operado a una velocidad de 400 rpm, 30% OD, 30 gSac/L, en oscuridad continua y a 25 °C. Bajo estas condiciones se registró la mejor actividad antiplasmodial de toda la investigación e incluso, se considera promisoria (IC50 = 10.67 µg/mL). Además, este extracto inhibe el parásito en el estadío trofozoíto inmaduro temprano. Finalmente, se identificaron tentativamente 16 metabolitos dentro del extracto, entre limonoides, diterpenoides y derivados de ácidos grasos. De esta forma, se logró establecer condiciones adecuadas para los cultivos celulares de Azadirachta indica que promueven la producción de metabolitos secundarios con actividad antiplasmodial in vitro sobre la cepa FCR3 de Plasmodium falciparum, siendo dicha actividad promisoria y permitiendo perfilar el cultivo de células en suspensión de neem como una nueva alternativa de producción de antimaláricos. |
dc.description.abstract | Malaria causes around 500,000 deaths worldwide each year. It is caused by parasites of the Plasmodium genus, such as Plasmodium falciparum, that has acquired resistance to current medications; thus, forcing a continuous search for new bioactive substances to achieve its control. In this research, conditions for Azadirachta indica cell cultures were established to stimulate the production of secondary metabolites with in vitro antiplasmodial activity against Plasmodium falciparum (FCR3 strain). Extracts were obtained from the biomass cultivated under those conditions and bioactivity assays were carried out. Additionally, the most active extract was evaluated in a Stage-Specific Timing and also analyzed using UPLC-ESI-Qtof-MS in order to detect some compounds probably involved in antiplasmodial activity. The best culture conditions using shake flasks were: 25° C, 40 gSac / L, no addition of hydrogen peroxide and no baffles (IC50 = 26.16 µg / mL), and 15° C, using MS medium, adding sodium acetate precursor, not using elicitors and 30 gSac / L) (IC50 = 18.72 µg / mL, the best activity in this scale). However, in vitro antiplasmodial activity was mainly promoted in a stirred tank bioreactor operated at 400 rpm, 30% DO, 30 gSac / L, in continuous darkness and 25° C. The best antiplasmodial activity was enriched under these conditions and is even considered as promising (IC50 = 10.67 µg / mL). Furthermore, this extract inhibits the parasite in the early immature trophozoite stage. Finally, 16 metabolites were tentatively identified within the extract, including limonoids, diterpenoids, and fatty acid derivatives. In this way, it was possible to establish conditions for Azadirachta indica cell cultures that promote the production of secondary metabolites with in vitro antiplasmodial activity against Plasmodium falciparum (FCR3 strain). This activity is promising and sets the cells suspension cultures of neem as a new alternative to the production of antimalarials. |
dc.description.sponsorship | Colciencias |
dc.format.extent | 115 |
dc.format.mimetype | application/pdf |
dc.language.iso | spa |
dc.rights | Derechos reservados - Universidad Nacional de Colombia |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/4.0/ |
dc.subject.ddc | Biotecnología |
dc.title | Cultivo de células en suspensión de Azadirachta indica A. Juss para la producción de metabolitos secundarios con actividad antiplasmodial |
dc.title.alternative | Azadirachta indica A. Juss cell cultures to produce secondary metabolites with antiplasmodial activity |
dc.type | Otro |
dc.rights.spa | Acceso abierto |
dc.type.driver | info:eu-repo/semantics/other |
dc.type.version | info:eu-repo/semantics/acceptedVersion |
dc.publisher.program | Medellín - Ciencias - Doctorado en Biotecnología |
dc.contributor.corporatename | Universidad Nacional de Colombia - Sede Medellín |
dc.description.degreelevel | Doctorado |
dc.publisher.department | Escuela de biociencias |
dc.publisher.branch | Universidad Nacional de Colombia - Sede Medellín |
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dc.rights.accessrights | info:eu-repo/semantics/openAccess |
dc.subject.proposal | Antimalarial |
dc.subject.proposal | Antimalárico |
dc.subject.proposal | Half maximal inhibitory concentration (IC50) |
dc.subject.proposal | Biorreactor de tanque agitado |
dc.subject.proposal | Concentración inhibitoria 50 (IC50) limonoides |
dc.subject.proposal | Limonoids |
dc.subject.proposal | Plasmodium falciparum |
dc.subject.proposal | Malaria |
dc.subject.proposal | Stirred tank bioreactor |
dc.subject.proposal | Neem |
dc.subject.proposal | Plasmodium falciparum |
dc.type.coar | http://purl.org/coar/resource_type/c_1843 |
dc.type.coarversion | http://purl.org/coar/version/c_ab4af688f83e57aa |
dc.type.content | Text |
oaire.accessrights | http://purl.org/coar/access_right/c_abf2 |
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