The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide

Perinatal asphyxia (PA) is a clinical condition characterized by oxygen deprivation, following PA brain damage progresses, leading to behavioural cognitive disabilities affecting surviving neonates. There is still lack of consensus on suitable therapeutic approaches and protocols preventing the prog...

Descripción completa

Detalles Bibliográficos
Autor: Lespay Rebolledo, Carolyne Andrea
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2019
País:Chile
OAI Identifier:oai:repositorio.anid.cl:10533/241975
Acceso en línea:https://hdl.handle.net/10533/241975
Access Level:acceso abierto
Palabra clave:Medicina y Ciencias de la Salud
Otras Ciencias Médicas
id CL_d4dd5f562061eea8af2bdbb4ff7ccc71
oai_identifier_str oai:repositorio.anid.cl:10533/241975
network_acronym_str CL
network_name_str Chile
repository_id_str
dc.title.es_CL.fl_str_mv The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
title The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
spellingShingle The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
Lespay Rebolledo, Carolyne Andrea
Medicina y Ciencias de la Salud
Otras Ciencias Médicas
Otras Ciencias Médicas
title_short The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
title_full The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
title_fullStr The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
title_full_unstemmed The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
title_sort The progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamide
dc.creator.none.fl_str_mv Lespay Rebolledo, Carolyne Andrea
author Lespay Rebolledo, Carolyne Andrea
author_facet Lespay Rebolledo, Carolyne Andrea
author_role author
dc.contributor.advisor.none.fl_str_mv Herrera-Marschitz, Mario Guillermo
dc.contributor.institution.es_CL.fl_str_mv UNIVERSIDAD DE CHILE
dc.subject.oecd1n.es_CL.fl_str_mv Medicina y Ciencias de la Salud
topic Medicina y Ciencias de la Salud
Otras Ciencias Médicas
Otras Ciencias Médicas
dc.subject.oecd2n.es_CL.fl_str_mv Otras Ciencias Médicas
dc.subject.oecd3n.es_CL.fl_str_mv Otras Ciencias Médicas
description Perinatal asphyxia (PA) is a clinical condition characterized by oxygen deprivation, following PA brain damage progresses, leading to behavioural cognitive disabilities affecting surviving neonates. There is still lack of consensus on suitable therapeutic approaches and protocols preventing the progression of neuronal damage. Reduction of oxidative stress is one of the identified molecular targets, based on the fact that the immature brain is highly susceptible to free radicals, showing scarcely developed antioxidant defences. In the present thesis it was proposed the hypothesis that progression of brain damage during a delayed cell death period in vulnerable brain areas of animals exposed to perinatal asphyxia is associated with sustained oxidative stress along development, impairing redox homeostasis. Nicotinamide enhances the response of glutathione-dependent enzymes by a pentose phosphate dependent pathway, preventing asphyxia-dependent brain damage. The hypothesis was evaluated using a rat model of severe perinatal asphyxia. The first part of the present Thesis evaluated the effect of global perinatal asphyxia on several parameters of oxidative stress and cell death in rat brain tissue sampled at an extended neonatal period of 14 days. Brain samples (mesencephalon, telencephalon and hippocampus) were assayed for glutathione (reduced and oxidized levels; spectrophotometry), tissue reducing capacity (potassium ferricyanide reducing assay, FRAP), catalase (the key enzyme protecting against oxidative stress and reactive oxygen species, Western blots and ELISA) and cleaved caspase-3 (the key executioner of apoptosis, Western blots) levels. It was found that global PA produced a regionally specific and sustained increase in GSSG/GSH ratio, a regionally specific decrease in tissue reducing capacity and a regionally and time specific decrease of catalase activity and increase of cleaved caspase-3 levels, indicating a long-term impairment in redox homeostasis. Multivariate analysis demonstrated that progression of brain damage along postnatal days is importantly influenced by changes on glutathione ratio, cleaved caspase-3 levels and catalase activity. In the second part of the Thesis, the experimental approach focused on the damage induced in hippocampus at P1 and P14. Glutathione, glutathione reductase (GR), glutathione peroxydase (GPx) (all by spectrophotometry), catalase (Western blots and ELISA), TIGAR (Western blots), calpain (fluorescence), and XRCC1 (Western blots) were assayed for characterizing the glutathione-dependent redox pathway and for determining the effect of nicotinamide; a precursor of NAD+ and NADP+, on death cell mechanisms in hippocampus. It was found that global PA produced (i) a sustained increase of GSSG levels and GSSG/GSH ratio at P1 and P14; (ii) a decrease of GR, GPx, and catalase activity at P1 and P14; (iii) a decrease at P1, followed by an increase at P14 of TIGAR levels; (iv) an increase of calpain activity at P14; and (v) an increase of XRCC1 levels, but only at P1. (vi) Nicotinamide prevented the effect of PA on GSSG levels and GSSG/GSH ratio, and on GR, GPx, and catalase activity, also on the enhancement of TIGAR levels and calpain activity observed at P14. The present thesis demonstrates that PA induces a sustained oxidative stress along development in mesencephalon and hippocampus, both vulnerable brain areas of animals exposed to PA, whereas telencephalon, a resistant brain area, displayed an increasing of antioxidant response mediated by glutathione and FRAP. The progression of brain damage during a delayed cell death period is associated mainly with glutathione ratio, cleaved caspase-3 levels and catalase activity. Nicotinamide enhanced the response of glutathione-dependent enzymes by a pentose phosphate dependent pathway, preventing asphyxia-dependent brain damage by a reduction of death cell mechanisms mediated by calpain. In conclusion the present thesis demonstrates that changes in redox environment induced by PA are sustained along the time, contributing to progression of brain damage, but also resistance to further damage during the delayed death cell period. The changes in redox environment can be modulated, controlling the cell metabolism induced by pentose phosphate dependent pathways.
publishDate 2019
dc.date.issued.es_CL.fl_str_mv 2019
dc.date.accessioned.none.fl_str_mv 2020-06-15T15:28:42Z
2022-08-16T20:03:56Z
dc.date.available.none.fl_str_mv 2020-06-15T15:28:42Z
2022-08-16T20:03:56Z
dc.type.driver.es_CL.fl_str_mv info:eu-repo/semantics/doctoralThesis
dc.type.openaire.none.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.tesis.none.fl_str_mv Tesis
format doctoralThesis
status_str publishedVersion
dc.identifier.folio.es_CL.fl_str_mv 21140281
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/10533/241975
identifier_str_mv 21140281
url https://hdl.handle.net/10533/241975
dc.relation.none.fl_str_mv instname: Conicyt
reponame: Repositorio Digital RI2.0
dc.relation.projectid.es_CL.fl_str_mv info:eu-repo/grantAgreement//21140281
dc.relation.set.none.fl_str_mv info:eu-repo/semantics/dataset/hdl.handle.net/10533/93488
dc.relation.uri.es_CL.fl_str_mv http://repositorio.uchile.cl/bitstream/handle/2250/173585/Progressive-brain-damage-observed-following-perinatal-asphyxia-is-sustained-by-a-long-term-impairment-of-redox-homeostasis.pdf?sequence=1&isAllowed=y
dc.rights.es_CL.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.uri.*.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/cl/
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/cl/
bitstream.url.fl_str_mv https://repositorio.anid.cl/bitstreams/d366fe4d-34a3-41db-8524-c777eb03236b/download
https://repositorio.anid.cl/bitstreams/dad1cbf3-a950-4846-b0ae-5141051e1c2a/download
https://repositorio.anid.cl/bitstreams/ce02910a-224a-4c42-ad48-79144ff18d14/download
https://repositorio.anid.cl/bitstreams/9d58d5d3-b259-44de-a96a-c7e7ac339d8a/download
https://repositorio.anid.cl/bitstreams/54653267-7032-4140-a8f9-4ef91adefc34/download
bitstream.checksum.fl_str_mv 593a6e7305c66c56041a9f9e15a649c1
9761646f553f57cccf9e902ea18318be
f97bcfdf58f3e17b5cec231112dab5b1
76194014d11f4d3af085087b2262d489
1c03a04a7811ae0728ccc34059450028
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositorio ANID
repository.mail.fl_str_mv aletelier@anid.cl
_version_ 1843518758929301504
spelling UNIVERSIDAD DE CHILELespay Rebolledo, Carolyne Andrea2019https://hdl.handle.net/10533/241975http://purl.org/coar/access_right/c_abf2Otras Ciencias MédicasOtras Ciencias MédicasMedicina y Ciencias de la SaludThe progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamideHerrera-Marschitz, Mario GuillermoUNIVERSIDAD DE CHILEChileLespay Rebolledo, Carolyne Andrea2020-06-15T15:28:42Z2022-08-16T20:03:56Z2020-06-15T15:28:42Z2022-08-16T20:03:56Z2019Perinatal asphyxia (PA) is a clinical condition characterized by oxygen deprivation, following PA brain damage progresses, leading to behavioural cognitive disabilities affecting surviving neonates. There is still lack of consensus on suitable therapeutic approaches and protocols preventing the progression of neuronal damage. Reduction of oxidative stress is one of the identified molecular targets, based on the fact that the immature brain is highly susceptible to free radicals, showing scarcely developed antioxidant defences. In the present thesis it was proposed the hypothesis that progression of brain damage during a delayed cell death period in vulnerable brain areas of animals exposed to perinatal asphyxia is associated with sustained oxidative stress along development, impairing redox homeostasis. Nicotinamide enhances the response of glutathione-dependent enzymes by a pentose phosphate dependent pathway, preventing asphyxia-dependent brain damage. The hypothesis was evaluated using a rat model of severe perinatal asphyxia. The first part of the present Thesis evaluated the effect of global perinatal asphyxia on several parameters of oxidative stress and cell death in rat brain tissue sampled at an extended neonatal period of 14 days. Brain samples (mesencephalon, telencephalon and hippocampus) were assayed for glutathione (reduced and oxidized levels; spectrophotometry), tissue reducing capacity (potassium ferricyanide reducing assay, FRAP), catalase (the key enzyme protecting against oxidative stress and reactive oxygen species, Western blots and ELISA) and cleaved caspase-3 (the key executioner of apoptosis, Western blots) levels. It was found that global PA produced a regionally specific and sustained increase in GSSG/GSH ratio, a regionally specific decrease in tissue reducing capacity and a regionally and time specific decrease of catalase activity and increase of cleaved caspase-3 levels, indicating a long-term impairment in redox homeostasis. Multivariate analysis demonstrated that progression of brain damage along postnatal days is importantly influenced by changes on glutathione ratio, cleaved caspase-3 levels and catalase activity. In the second part of the Thesis, the experimental approach focused on the damage induced in hippocampus at P1 and P14. Glutathione, glutathione reductase (GR), glutathione peroxydase (GPx) (all by spectrophotometry), catalase (Western blots and ELISA), TIGAR (Western blots), calpain (fluorescence), and XRCC1 (Western blots) were assayed for characterizing the glutathione-dependent redox pathway and for determining the effect of nicotinamide; a precursor of NAD+ and NADP+, on death cell mechanisms in hippocampus. It was found that global PA produced (i) a sustained increase of GSSG levels and GSSG/GSH ratio at P1 and P14; (ii) a decrease of GR, GPx, and catalase activity at P1 and P14; (iii) a decrease at P1, followed by an increase at P14 of TIGAR levels; (iv) an increase of calpain activity at P14; and (v) an increase of XRCC1 levels, but only at P1. (vi) Nicotinamide prevented the effect of PA on GSSG levels and GSSG/GSH ratio, and on GR, GPx, and catalase activity, also on the enhancement of TIGAR levels and calpain activity observed at P14. The present thesis demonstrates that PA induces a sustained oxidative stress along development in mesencephalon and hippocampus, both vulnerable brain areas of animals exposed to PA, whereas telencephalon, a resistant brain area, displayed an increasing of antioxidant response mediated by glutathione and FRAP. The progression of brain damage during a delayed cell death period is associated mainly with glutathione ratio, cleaved caspase-3 levels and catalase activity. Nicotinamide enhanced the response of glutathione-dependent enzymes by a pentose phosphate dependent pathway, preventing asphyxia-dependent brain damage by a reduction of death cell mechanisms mediated by calpain. In conclusion the present thesis demonstrates that changes in redox environment induced by PA are sustained along the time, contributing to progression of brain damage, but also resistance to further damage during the delayed death cell period. The changes in redox environment can be modulated, controlling the cell metabolism induced by pentose phosphate dependent pathways.21140281https://hdl.handle.net/10533/241975instname: Conicytreponame: Repositorio Digital RI2.0info:eu-repo/grantAgreement//21140281info:eu-repo/semantics/dataset/hdl.handle.net/10533/93488http://repositorio.uchile.cl/bitstream/handle/2250/173585/Progressive-brain-damage-observed-following-perinatal-asphyxia-is-sustained-by-a-long-term-impairment-of-redox-homeostasis.pdf?sequence=1&isAllowed=yinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/3.0/cl/Medicina y Ciencias de la SaludOtras Ciencias MédicasOtras Ciencias MédicasThe progressive brain damage observed following perinatal asphyxa is sustained by a long-term impairment of redox homeostasis: effect of nicotinamideinfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionTesisTesishttps://hdl.handle.net/10533/241975f3102eca-a7ca-4626-8d05-977034825eb6virtual::57610-1f3102eca-a7ca-4626-8d05-977034825eb6virtual::57610-1LICENSElicense.txttext/plain1779https://repositorio.anid.cl/bitstreams/d366fe4d-34a3-41db-8524-c777eb03236b/download593a6e7305c66c56041a9f9e15a649c1MD51ORIGINALTesis C Lespay R.pdfThesisapplication/pdf5536519https://repositorio.anid.cl/bitstreams/dad1cbf3-a950-4846-b0ae-5141051e1c2a/download9761646f553f57cccf9e902ea18318beMD52CC-LICENSElicense_rdfapplication/octet-stream1232https://repositorio.anid.cl/bitstreams/ce02910a-224a-4c42-ad48-79144ff18d14/downloadf97bcfdf58f3e17b5cec231112dab5b1MD53TEXTTesis C Lespay R.pdf.txtExtracted texttext/plain278081https://repositorio.anid.cl/bitstreams/9d58d5d3-b259-44de-a96a-c7e7ac339d8a/download76194014d11f4d3af085087b2262d489MD54THUMBNAILTesis C Lespay R.pdf.jpgIM Thumbnailimage/jpeg4002https://repositorio.anid.cl/bitstreams/54653267-7032-4140-a8f9-4ef91adefc34/download1c03a04a7811ae0728ccc34059450028MD5510533/241975oai:repositorio.anid.cl:10533/2419752023-07-24 04:03:12.165http://creativecommons.org/licenses/by-nc-nd/3.0/cl/info:eu-repo/semantics/openAccesshttps://repositorio.anid.clRepositorio ANIDaletelier@anid.clTGljZW5jaWEgZGUgRGlzdHJpYnVjacOzbiBObyBFeGNsdXNpdmEKCkFsIGZpcm1hciB5IHByZXNlbnRhciBlc3RhIGxpY2VuY2lhICwgdXN0ZWQgKGVsIGF1dG9yKGVzKSBvIHRpdHVsYXIgZGUgZGVyZWNob3MgZGUgYXV0b3IpIGdhcmFudGl6YSBhIGxhIENvbWlzacOzbiBOYWNpb25hbCBkZSBJbnZlc3RpZ2FjacOzbiBDaWVudMOtdGljYSB5IFRlY25vbMOzZ2ljYSAoQ09OSUNZVCkgZWwgZGVyZWNobyBubyBleGNsdXNpdm8gZGUgcmVwcm9kdWNpciAsIHRyYWR1Y2lyIChjb21vIHNlIGRlZmluZSBtw6FzIGFiYWpvKSAsIHkvbyBkaXN0cmlidWlyIHN1IGRvY3VtZW50byAoaW5jbHV5ZW5kbyBlbCByZXN1bWVuKSBlbiB0b2RvIGVsIG11bmRvIGVuIGZvcm1hIGltcHJlc2EgeSBlbiBmb3JtYXRvIGVsZWN0csOzbmljbyB5IGVuIGN1YWxxdWllciBtZWRpbywgaW5jbHV5ZW5kbywgcGVybyBubyBsaW1pdGFkbyBhLCBhdWRpbyBvIHbDrWRlby4KClVzdGVkIGFjZXB0YSBxdWUgQ09OSUNZVCBwdWVkZSwgc2luIGFsdGVyYXIgc3UgY29udGVuaWRvLCBjb252ZXJ0aXJsbyBhIGN1YWxxdWllciBtZWRpbyBvIGZvcm1hdG8gcGFyYSBlbCBmaW4gZGUgbGEgY29uc2VydmFjacOzbi4KClRhbWJpw6luIGFjZXB0YSBxdWUgQ09OSUNZVCBwdWVkZSB0ZW5lciBtw6FzIGRlIHVuYSBjb3BpYSBkZSBlc3RlIGRvY3VtZW50byBwYXJhIGZpbmVzIGRlIHNlZ3VyaWRhZCwgY29waWFzIGRlIHNlZ3VyaWRhZCB5IGNvbnNlcnZhY2nDs24uCgpVc3RlZCBkZWNsYXJhIHF1ZSBlbCBkb2N1bWVudG8gZXMgdW4gdHJhYmFqbyBvcmlnaW5hbCwgeSBxdWUgdXN0ZWQgdGllbmUgZWwgZGVyZWNobyBkZSBvdG9yZ2FyIGxvcyBkZXJlY2hvcyBjb250ZW5pZG9zIGVuIGVzdGEgbGljZW5jaWEuIFRhbWJpw6luIGRlY2xhcmEgcXVlIHN1IHBldGljacOzbiBubywgYSBsbyBtZWpvciBkZSBzdSBjb25vY2ltaWVudG8sIGluZnJpbmdlIGxvcyBkZXJlY2hvcyBkZSBhdXRvciBkZSBuYWRpZS4KClNpIGVsIGRvY3VtZW50byBjb250aWVuZSBtYXRlcmlhbGVzIGRlIGxvcyBxdWUgbm8gdGllbmVuIGRlcmVjaG9zIGRlIGF1dG9yICwgdXN0ZWQgZGVjbGFyYSBxdWUgaGEgb2J0ZW5pZG8gZWwgcGVybWlzbyBzaW4gcmVzdHJpY2Npw7NuIGRlbCBwcm9waWV0YXJpbyBkZSBsb3MgZGVyZWNob3MgcmVxdWVyaWRvcyBwb3IgZXN0YSBsaWNlbmNpYSB5IHF1ZSBlc2UgbWF0ZXJpYWwgY3V5b3MgZGVyZWNob3Mgc29uIGRlIHRlcmNlcm9zIGVzdMOhIGNsYXJhbWVudGUgaWRlbnRpZmljYWRvIHkgcmVjb25vY2lkbyBlbiBlbCB0ZXh0byBvIGNvbnRlbmlkbyBkZWwgZG9jdW1lbnRvIGVudHJlZ2Fkby4KClNJIEVMIEVOVsONTyBTRSBCQVNBIEVOIEVMIFRSQUJBSk8gUVVFIEhBIFNJRE8gUEFUUk9OQURPIE8gQVBPWUFETyBQT1IgQUxHVU5BIElOU1RJVFVDScOTTiBRVUUgTk8gU0VBIENPTklDWVQsIFVTVEVEIEFDRVBUQSBRVUUgSEEgQ1VNUExJRE8gQSBDVUFMUVVJRVIgREVSRUNITyBERSBSRVZJU0nDk04gVSBPVFJBUyBPQkxJR0FDSU9ORVMgUkVRVUVSSURBUyBQT1IgRElDSE8gQ09OVFJBVE8gTyBBQ1VFUkRPLgoKQ09OSUNZVCBpZGVudGlmaWNhcsOhIGNsYXJhbWVudGUgc3Ugbm9tYnJlKGVzKSBjb21vIGVsIGF1dG9yKGVzKSBvIHByb3BpZXRhcmlvKHMpIGRlIGxhIHByZXNlbnRhY2nDs24gLCB5IG5vIGhhcsOhIG5pbmd1bmEgYWx0ZXJhY2nDs24sIGV4Y2VwdG8gc2Vnw7puIGxvIHBlcm1pdGlkbyBwb3IgbGEgbGljZW5jaWEsIHBhcmEgc3UgcHJlc2VudGFjacOzbi4K
score 15,812429

Similares en LA Referencia