Orientadora: Prof. Dra. Giselle Cerchiaro / Tese (doutorado) - Universidade Federal do ABC. Programa de Pós-Graduação em Ciência e Tecnologia/Química, 2017. / Fortes evidencias sugerem que a deterioracao da homeostase de metais essenciais, o aumento de danos oxidativos a biomoleculas e o aumento de especies oxidantes estao estreitamente relacionados ao desencadeamento, manutencao e evolucao do quadro de algumas doencas neurodegenerativas como Alzheimer, Parkinson e eslerose lateral amiotrofica. Apesar desta relacao estar bem estabelecida, ainda e desconhecida quais os mecanismos metabolicos relacionados a tais patologias. Tratando de diferentes doencas neurodegenerativas, com origens e mecanismos diversos, para que se possa propor uma prevencao e/ou tratamento para pacientes acometidos por sintomas seriamente debilitantes, e necessario fazer uma analise multidimensional, relacionando dados quimicos e comportamentais. Este trabalho mostrou os efeitos deleterios da inducao do estresse oxidativo por dois tratamentos diferentes (H2O2 e ¿À-amiloide na forma oligomerica) em linhagens celulares relacionadas ao sistema nervoso central (NSC-34, mHipoE-18, SH-SY5Y diferenciada e astrocitos imortalizados). Os dois tratamentos foram capazes de induzir o aumento da fosforilacao da proteina Tau, fator esse que esta diretamente ligado a doencas neurodegenerativas. De forma isolada, foram planejados experimentos que oferecem uma leitura da relacao dos danos celulares causados pelo fluxo de elementos essenciais (Fe, Cu, Zn, Ca, Se, Co, Mn, Mg e Cr) e os danos a biomoleculas (peroxidacao lipidica, carbonilacao de proteinas e danos ao DNA). O fluxo de elementos essenciais ocasionado por danos oxidativos nas mitocondrias e no nucleo foram analisados separadamente, bem como o efeito dos diferentes elementos essenciais na celula integra. Como era esperado, cada tratamento levou a mecanismos de resposta distintos para cada linhagem celular. No tempo de 24 h os dois tratamentos induziram danos ao DNA para todas as linhagens celulares estudadas. Ja para a peroxidacao lipidica, o tratamento com H2O2 danificou lipideos da linhagem SHSY5Y e diminuiu os danos lipidicos para a linhagem de astrocitos imortalizados enquanto que o tratamento com A¿ÀO so aumentou a peroxidacao lipidica para as celulas da linhagem NSC-34. Alem disso, o tratamento de H2O2 induziu danos proteinas nas celulas das linhagens NSC-34 e SHSY5Y, enquanto que o tratamento com A¿ÀO induziu um aumento dos danos a proteinas nas celulas da linhagem NSC-34 e uma diminuicao destes danos nas celulas SH-SY5Y. O tratamento com H2O2 induziu o aumento de Zn e Co e uma diminuicao de Mg e Se no nucleo; diminuicao de Zn, Ca, Se e Mn na mitocondria e diminuicao de Zn, Mn e Mg em celulas da linhagem NSC-34. Em celulas da linhagem SH-SY5Y o tratamento com H2O2 induziu o aumento de Ca na celula total, aumento de Co e diminuicao de Ca no nucleo e aumento de Fe, Zn e Ca na mitocondria. De modo geral, podemos observar um aumento do fluxo de Fe em todas as linhagens submetidas aos tratamentos oxidativos. Verificamos que o aumento de Co no nucleo pode ser utilizado como um biomarcador do efeito deleterio do estresse oxidativo induzido por H2O2 por aparecer como resposta a este tratamento em todas as linhagens estudadas nesse trabalho. Entretanto para o tratamento com A¿ÀO nenhum biomarcador foi encontrado uma vez que as diferentes linhagens estudadas tiveram alteracoes na homeostase de elementos de maneira distinta. / Strong evidence suggests that the deterioration of essential metal homeostasis, increased oxidative damage to biomolecules, and increased oxidant species are closely related to the onset, maintenance, and evolution of neurodegenerative diseases such as Alzheimer's, Parkinson's, and Amyotrophic lateral sclerosis. Although this relationship is well established, the underlying metabolic mechanisms behind these pathologies remains unknown. To develop appropriate treatments for different neurodegenerative diseases with diverse origins and mechanisms, so that a prevention and/or treatment can be proposed for patients suffering from severely debilitating symptoms, it is necessary to perform a multidimensional analysis, relating chemical, metabolomic and behavioral data. This study makes an essential contribution to this field, as we have collected relevant data on the deleterious effects of oxidative stress induction by two different treatments (H2O2 and AâO) on cell lines related to the central nervous system (NSC-34, mHipoE-18, differentiated SH-SY5Y and immortalized astrocytes). Both oxidative treatments demonstrated the ability to induce an increase in Tau phosphorylation, a factor that is directly related to neurodegenerative disease. Experiments were carried out in isolation to evaluate the relationship between cellular damage caused by the flow of essential elements (Fe, Cu, Zn, Ca, Se, Co, Mn, Mg, and Cr) and damage to biomolecules (lipid peroxidation, Carbonylation of proteins and DNA damage). The flow of essential elements caused by oxidative damage in the mitochondria and nucleus were analyzed separately, as well as the effect of different essential elements within the whole cell. As expected, each treatment led to distinct response mechanisms for each cell line. At 24 h, both treatments induced DNA damage for all cell lines studied. Analyzing lipid peroxidation, treatment with H2O2 increased lipid damage to the SHSY5Y cell line strain and decreased lipid damage to the immortalized astrocyte lineage, whereas Aâ treatment only increased lipid peroxidation for NSC-34 cells. In addition, H2O2 treatment induced protein damage in NSC-34 and SHSY5Y cell lines, whereas AâO treatment induced an increase in protein damage in NSC-34 lineage cells and a decrease in damage to SH-SY5Y cell line. Treatment with H2O2 induced increased Zn and Co and decreased Mg and Se in the nucleus, decreased Zn, Ca, Se and Mn in the mitochondria and decreased Zn, Mn, and Mg in NSC-34 lineage cells. In cells from the SH-SY5Y line, treatment with H2O2 induced the increase of Ca in the total cell, an increase in Co and a decrease in Ca of the nucleus, and the increase of Fe, Zn and Ca in mitochondria. For cells of the immortalized astrocyte line, when treated with H2O2 there was an increase of Fe in the total cell, an increase of Cu, Co and Cr in the nucleus and increased Fe and Cu in mitochondria. However, Aâ treatment to cells of the NSC-34 lineage induced a decrease in Mn and Mg in the total cell, an increase of Fe, Cu, Zn, Ca and Cr in the nucleus and a decrease of Ca, Se and Mn in mitochondria. In cells from the SH-SY5Y strain, AâO treatment induced increased Fe and Ca in the total cell, decreased Ca in the nucleus and increased Fe, Cu, Zn, Ca, Se and Co in the mitochondria. Finally, for immortalized astrocytic lineage cells treated with AâO, there was increased Fe and Cr both in the total cell and the nucleus, and increased Fe in mitochondria. In this way, we verified that increased Co in the nucleus can be used as a biomarker of the deleterious effects of H2O2 induced oxidative stress, as all cell lines displayed a convergent response. However, no such biomarker could be identified for treatment with AâO, as the elemental homeostatic responses observed were varied across different cell lines.