Efeito da manipulação do microambiente neuronal indiretamente através da pinealectomia ou transplante de células derivadas de medula óssea nos modelos animais da Doença de Parkinson induzido por neurotoxinas

Orientadora : Profa. Dra. Maria Aparecida Barbato Frazão VitalCo-orientadora : Profª Drª Valdo José Dias da SilvaTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia. Defesa: Curitiba,17/09/2014Inclui referênciasÁrea de concentração: FarmacologiaResumo: A Doença de Parkinson é uma doença neurodegenerativa caracterizada pela perda de neurônios dopaminérgicos na SNpc. O presente trabalho visou investigar em duas linhas distintas o papel do microambiente celular na neurodegeneração. Na primeira linha de investigação, os animais foram avaliados quanto ao efeito da pinealectomia sobre a neurodegeneração e estresse oxidativo induzido pela administração de MPTP e 6-OHDA. A administração dessas neurotoxinas resultou em comprometimento motor em ambos os grupos Sham e Px 24 h após a lesão no teste do campo aberto. Contudo, 7 e 14 dias após a cirurgia, os animais lesados pertencentes ao grupo Sham apresentavam recuperação motora, ao contrário dos grupos Px e lesados. No teste de natação forçada os animais lesados apresentavam menor tempo de natação em relação aos grupos controles, e ainda, os grupos Sham- 6-OHDA e Px-6-OHDA apresentaram maior tempo de imobilidade comparado aos controles. Corroborando com essas alterações, significativa degeneração dopaminérgica foi evidenciada nos animais lesados, e ainda, aumento na quantidade de células contendo ROS nesses grupos em relação aos grupos controles. Já no segundo estudo, o modelo do MPTP foi empregado para avaliar o efeito do transplante de BMMC ou BM-MSC em diferentes tempos, imediatamente e 24 h após a infusão da neurotoxina. A infusão de MPTP resultou em quebra da BHE e prejuízo motor 24 h após a infusão de MPTP, e o tratamento com BMMC imediatamente após a lesão aumentou significativamente a perda de células TH-ir nesses animais, ao contrário do tratamento com BM-MSCs. Por outro lado, o tratamento com BMMC 24 h após a infusão de MPTP resultou em similar perda de neurônios TH-ir comparado ao grupo MPTP-salina. No teste de natação forçada, o grupo MPTP-BMMC apresentou aumento no tempo de imobilidade comparado aos grupos Sham e MPTP, e ainda, aumento no número de células CD45+ e micróglia ativada nas secções estudadas. O presente estudo sugere que o transplante de BM-MSC em animais lesados com MPTP module positivamente o microambiente neuronal favorecendo a sobrevivência neuronal, enquanto que o transplante com BMMCs e a redução dos níveis fisiológicos de melatonina mostraram-se desfavorável ao microambiente neuronal. Palavras-chave: MPTP, 6-OHDA, BMMC, BM-MSC, pinealectomia e DP.Abstract: Parkinson's disease (PD) is a neurodegenerative disease characterized by loss of dopaminergic neurons in the SNpc. This study aimed to investigate the role of cellular microenvironment on the neurodegenration in two distinct investigation lines. In the first line, the animals were evaluated for the effect of pinealectomy (Px) on neurodegeneration and oxidative stress induced by MPTP or 6-OHDA. The administration of these neurotoxins results in motor impairment in both sham and Px groups 24 h after injury in the open field test. However, 7 and 14th days after injury, the injured animals belonging to Sham group showed recovery of motor abnormalities, unlike injured animals of the Px-groups. In the forced swim test, the injured animals exhibited shorter swimming compared to control groups, and futher, Sham-6-OHDA and Px-6-OHDA groups showed a significant increase in immobility time compared to control groups. Supporting these alterations, significant dopaminergic loss in animals injured was observed in comparison to control groups, and futher, increased quantity of cellular ROS in these groups. In the second study, the MPTP model was used to evaluate the effect of BMMC and BM-MSCs at different times, immediately and 24 hours, after neurotoxins infusion. The MPTP infusion resulted in breakdown of bloodbrain barrier and motor impairment 24h after MPTP infusion, and the BMMC treatment immediately after lesion induced a significant increase loss of TH-ir neurons in these animals when compared to MPTP-saline group, in opposite to the BM-MSCs treatment. In the other hand, MPTP-lesioned rats treated with BMMCs 24 h after to neurotoxin infusion showed similar loss of dopaminergic neurons to MPTP-saline. In the forced swimming test, MPTP-BMMC treated group presented an increase in the immobility time compared to sham and MPTP-saline group, and futher increase in the number of CD45-labeled cells and activated microglial cells in the sections studied. This study suggests that the BM-MSCs transplantation in MPTP-lesioned rats positively modulate the microenvironment favoring neuronal neuronal survival, whereas transplantation BMMCs and the reduction of physiological levels of melatonin have proved unfavorable neuronal microenvironment. Keywords: MPTP, 6-OHDA, BMMC, BM-MSC, pinealectomy e PD

Efeito da melatonina em modelo animal de parkinsonismo induzido pelo MPTP /

Orientadora : Maria Aparecida B.F. VitalDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia. Defesa: Curitiba, 2007Inclui bibliografi

Efeito da melatonina em modelo animal de parkinsonismo induzido pelo MPTP /

Orientadora : Maria Aparecida B.F. VitalDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia. Defesa: Curitiba, 2007Inclui bibliografi

Serotoninergic Modulation of Basal Cardiovascular Responses and Responses Induced by Isotonic Extracellular Volume Expansion in Rats

Background: Isotonic blood volume expansion (BVE) induced alterations of sympathetic and parasympathetic activity in the heart and blood vessels, which can be modulated by serotonergic pathways. Objective: To evaluate the effect of saline or serotonergic agonist (DOI) administration in the hypothalamic paraventricular nucleus (PVN) on cardiovascular responses after BVE. Methods: We recorded pulsatile blood pressure through the femoral artery to obtain the mean arterial pressure (MAP), systolic (SBP) and diastolic blood pressure (DBP), heart rate (HR) and the sympathetic-vagal ratio (LF/HF) of Wistar rats before and after they received bilateral microinjections of saline or DOI into the PVN, followed by BVE. Results: No significant differences were observed in the values of the studied variables in the different treatments from the control group. However, when animals are treated with DOI followed by BVE there is a significant increase in relation to the BE control group in all the studied variables: MBP (114.42 +/- 7.85 vs 101.34 +/- 9.17)SBP (147.23 +/- 14.31 vs 129.39 +/- 10.70)DBP (98.01 +/- 4.91 vs 87.31 +/- 8.61)HR (421.02 +/- 43.32 vs 356.35 +/- 41.99)and LF/HF ratio (2.32 +/- 0.80 vs 0.27 +/- 0.32). Discussion: The present study showed that the induction of isotonic BVE did not promote alterations in MAP, HR and LF/HF ratio. On the other hand, the injection of DOI into PVN of the hypothalamus followed by isotonic BVE resulted in a significant increase of all variables. Conclusion: These results suggest that serotonin induced a neuromodulation in the PVN level, which promotes an inhibition of the baroreflex response to BVE. Therefore, the present study suggests the involvement of the serotonergic system in the modulation of vagal reflex response at PVN in the normotensive rats.Univ Fed Triangulo Mineiro, Uberaba, MG, BrazilUniv Fed Sao Paulo UNIFESP, Sao Paulo, SP, BrazilUniv Fed Sao Paulo UNIFESP, Sao Paulo, SP, BrazilWeb of Scienc

Serotoninergic Modulation of Basal Cardiovascular Responses and Responses Induced by Isotonic Extracellular Volume Expansion in Rats

Abstract Background: Isotonic blood volume expansion (BVE) induced alterations of sympathetic and parasympathetic activity in the heart and blood vessels, which can be modulated by serotonergic pathways. Objective: To evaluate the effect of saline or serotonergic agonist (DOI) administration in the hypothalamic paraventricular nucleus (PVN) on cardiovascular responses after BVE. Methods: We recorded pulsatile blood pressure through the femoral artery to obtain the mean arterial pressure (MAP), systolic (SBP) and diastolic blood pressure (DBP), heart rate (HR) and the sympathetic-vagal ratio (LF/HF) of Wistar rats before and after they received bilateral microinjections of saline or DOI into the PVN, followed by BVE. Results: No significant differences were observed in the values of the studied variables in the different treatments from the control group. However, when animals are treated with DOI followed by BVE there is a significant increase in relation to the BE control group in all the studied variables: MBP (114.42±7.85 vs 101.34±9.17); SBP (147.23±14.31 vs 129.39±10.70); DBP (98.01 ±4.91 vs 87.31±8.61); HR (421.02±43.32 vs 356.35±41.99); and LF/HF ratio (2.32±0.80 vs 0.27±0.32). Discussion: The present study showed that the induction of isotonic BVE did not promote alterations in MAP, HR and LF/HF ratio. On the other hand, the injection of DOI into PVN of the hypothalamus followed by isotonic BVE resulted in a significant increase of all variables. Conclusion: These results suggest that serotonin induced a neuromodulation in the PVN level, which promotes an inhibition of the baroreflex response to BVE. Therefore, the present study suggests the involvement of the serotonergic system in the modulation of vagal reflex response at PVN in the normotensive rats

Endogenous resident c-Kit cardiac stem cells increase in mice with an exercise-induced, physiologically hypertrophied heart

Physical activity evokes well-known adaptations in the cardiovascular system. Although exercise training induces cardiac remodeling, whether multipotent stem cells play a functional role in the hypertrophic process remains unknown. To evaluate this possibility, C57BL/6 mice were subjected to swimming training aimed at achieving cardiac hypertrophy, which was morphologically and electrocardiographically characterized. Subsequently, c-Kit+Lin− and Sca-1+Lin− cardiac stem cells (CSCs) were quantified using flow cytometry while cardiac muscle-derived stromal cells (CMSCs, also known as cardiac-derived mesenchymal stem cells) were assessed using in vitro colony-forming unit fibroblast assay (CFU-F). Only the number of c-Kit+Lin− cells increased in the hypertrophied heart. To investigate a possible extracardiac origin of these cells, a parabiotic eGFP transgenic/wild-type mouse model was used. The parabiotic pairs were subjected to swimming, and the wild-type heart in particular was tested for eGFP+ stem cells. The results revealed a negligible number of extracardiac stem cells in the heart, allowing us to infer a cardiac origin for the increased amount of detected c-Kit+ cells. In conclusion, the number of resident Sca-1+Lin− cells and CMSCs was not changed, whereas the number of c-Kit+Lin− cells was increased during physiological cardiac hypertrophy. These c-Kit+Lin− CSCs may contribute to the physiological cardiac remodeling that result from exercise training

Effects of Cholinergic Stimulation with Pyridostigmine Bromide on Chronic Chagasic Cardiomyopathic Mice

Made available in DSpace on 2015-05-04T16:34:37Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) constança_brittoetal_IOC_2014.pdf: 3857100 bytes, checksum: 42bae205f256c445117f5bd53964bde3 (MD5) Previous issue date: 2014Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilUniversidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilUniversidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.University of Milan. L Sacco Hospital. Department of Clinical Sciences, Internal Medicine II. Milan, Italy.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.Universidade Federal do Triângulo Mineiro. Instituto de Ciências Biológicas e Naturais. Uberaba, MG, Brasil.The aim of the present study was to assess the effects of an anticholinesterase agent, pyridostigmine bromide (Pyrido), on experimental chronic Chagas heart disease in mice. To this end, male C57BL/6J mice noninfected (control:Con) or chronically infected (5 months) with Trypanosoma cruzi (chagasic:Chg) were treated or not (NT) with Pyrido for one month. At the end of this period, electrocardiogram (ECG); cardiac autonomic function; heart histopathology; serum cytokines; and the presence of blood and tissue parasites by means of immunohistochemistry and PCR were assessed. In NT-Chg mice, significant changes in the electrocardiographic, autonomic, and cardiac histopathological profiles were observed confirming a chronic inflammatory response. Treatment with Pyrido in Chagasic mice caused a significant reduction of myocardial inflammatory infiltration, fibrosis, and hypertrophy, which was accompanied by a decrease in serum levels of IFN with no change in IL-10 levels, suggesting a shift of immune response toward an anti-inflammatory profile. Lower nondifferent numbers of parasite DNA copies were observed in both treated and nontreated chagasic mice. In conclusion, our findings confirmthe marked neuroimmunomodulatory role played by the parasympathetic autonomic nervous systemin the evolution of the inflammatory-immune response to T. cruzi during experimental chronic Chagas heart disease in mice