Submitted by Izabel Franco (izabel-franco@ufscar.br) on 2016-09-15T13:00:14Z No. of bitstreams: 1 DissLGAP.pdf: 2357885 bytes, checksum: dd8d8d69a2f2f83808b3f1561307aefd (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-16T19:24:00Z (GMT) No. of bitstreams: 1 DissLGAP.pdf: 2357885 bytes, checksum: dd8d8d69a2f2f83808b3f1561307aefd (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-16T19:24:13Z (GMT) No. of bitstreams: 1 DissLGAP.pdf: 2357885 bytes, checksum: dd8d8d69a2f2f83808b3f1561307aefd (MD5) / Made available in DSpace on 2016-09-16T19:24:24Z (GMT). No. of bitstreams: 1 DissLGAP.pdf: 2357885 bytes, checksum: dd8d8d69a2f2f83808b3f1561307aefd (MD5) Previous issue date: 2015-10-02 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / It is well know that the respiratory network, undergoes significant development in the postnatal period. Among various processes, the maturing of the catecholaminergic (CA) system shows to be an important factor in the control and modulation of respiratory rhythmogenesis. Studies have also shown that these neurons are widely distributed in the Central Nervous System (CNS), including the A1/C1, A2/C2, C3, A5, A6 and A7 regions, having numerous projections to many regions of the brain. However, the development of respiratory network as well as its effect on the control of ventilation, is not yet fully understood. Thus, understanding the participation of CA neurons in the respiratory control during postnatal development is of most importance for a better understanding of some clinical disorders including Rett Syndrome, Sudden Infant Death Syndrome (SIDS) and Central Congenital Hypoventilation Syndrome (CCHS). Therefore, this study aimed to investigate the involvement of CA neurons in the brainstem on respiratory control in normoxic normocapnic, hypercapnic and hypoxic conditions during the postnatal period of male and female neonatal rats, through chemical injury with conjugated saporin anti-dopamine beta-hydroxylase (DBH-SAP). Thus, DBH-SAP (42 ng/100 nL – 1L), saporin (SAP – 1L) or phosphate buffered solution vehicle (PBS, 0.01M, pH 7.4 – 1L) were injected into the 4th ventricle in male and female neonates Wistar rats P0-1. Pulmonary ventilation ( EV ) was recorded in unanesthetized neonates (P7-8) by pressure plethysmography during normocapnia, hypercapnia (7% CO2) and hypoxia (10% O2) at 10 and 20 min after the start of exposure. Our data demonstrate that lesion of brainstem CA neurons increased ventilation in males and females newborn under room air conditions. In addition, the ventilatory response to hypercapnia was significantly reduced in male (57%) and female (55%) lesioned neonatal rats (Male – SAP group: 212.8 ± 7.0; PBS group: 203.9 ± 10.3; lesioned group: 151.1 ± 7.4; P < 0,001; Female – SAP group: 218.2 ± 10.4; PBS group: 200.0 ± 6.4; lesioned group: 154.0 ± 9.6; P < 0,001; all values relative to % of baseline). Also, a similar reduction was observed in the hypoxic condition (Male – SAP group: 185.2 ± 15.3; PBS group: 167.4 ± 5.0; lesioned group: 110.8 ± 9.2; P < 0,001; Female – SAP group: 197.3 ± 11.8; PBS group: 179.5 ± 13.7; lesioned: 129.4 ± 5.9; P < 0,001; all values relative to % of baseline). Additionally, the values for metabolic rate of control and lesioned groups, both males and females, did not differ significantly, whether in normoxic normocapnic, hypercapnic or hypoxic conditions. These results suggest that brainstem CA neurons exert a tonic inhibitory role in neonatal ventilation and promote an important excitatory modulation in CO2 and O2 chemosensitivity in unanesthetized males and females neonatal rats (P7-8). / Sabe-se que o sistema respiratório, bem como suas vias de controle, sofrem significativo desenvolvimento no período pós-natal. Dentre vários processos, o amadurecimento do sistema catecolaminérgico (CA) mostra-se como um importante fator no controle e modulação da ritmogênese respiratória. Estudos demonstram que esses neurônios estão amplamente distribuídos pelo Sistema Nervo Central (SNC), incluindo as regiões A1/C1, A2/C2, C3, A5, A6 e A7, e que apresentam inúmeras projeções para várias regiões do encéfalo. No entanto, a participação dos neurônios CA no controle respiratório durante o desenvolvimento pós-natal não está bem esclarecido, e esse entendimento é de extrema importância para uma melhor compreensão de alguns problemas clínicos que inclui a Síndrome de Rett, Síndrome da Morte Súbita Infantil (SIDS) e a Síndrome da Hipoventilação Central Congênita (CCHS). Sendo assim, o presente estudo teve por objetivo investigar a participação dos neurônios CA do tronco encefálico no controle respiratório em situações normóxica normocápnicas, hipercápnicas e hipóxicas durante o período pós-natal de ratas e ratos (P7-8), por meio de lesão química com saporina conjugada com anti-dopamina beta-hidroxilase (DBHSAP). Assim, DBH-SAP (42 ng/100 nL – 1L), Saporina (SAP – 1 L) ou veículo solução fosfato tamponado (PBS 0,01 M, pH 7,4 – 1 L) foram injetados no 4° ventrículo de ratas e ratos neonatos Wistar P0-1. A ventilação pulmonar ( EV ) foi registrada em neonatos não anestesiados (P7-8) por pletismografia de pressão, durante normóxia normocápnica, hipercapnia (7% CO2) e hipóxia (10% O2) aos 10 e 20 min após o início da exposição. Nossos dados demonstram que a lesão dos neurônios catecolaminérgicos do tronco encefálico promove um aumento da ventilação em neonatos machos e fêmeas durante a normóxia normocápnica. A resposta ventilatória à hipercapnia foi significativamente reduzida em ratos neonatos lesados (57%) e ratas (55%) (Machos – grupo SAP: 212,8 ± 7,0; grupo PBS: 203,9 ± 10,3; grupo lesado: 151,1 ± 7,4; P < 0,001; Fêmeas – grupo SAP: 218,2 ± 10,4; grupo PBS: 200,0 ± 6,4; grupo lesado: 154,0 ± 9,6; P < 0,001; todos os valores relativos à % do basal). Uma redução similar foi observada na resposta ventilatória à hipóxia (Machos – grupo SAP: 185,2 ± 15,3; grupo PBS: 167,4 ± 5,0; grupo lesado: 110,8 ± 9,2; P < 0,001; Fêmeas – grupo SAP: 197,3 ± 11,8; grupo PBS: 179,5 ± 13,7; grupo lesado: 129,4 ± 5,9; P < 0,001; todos os valores relativos à % do basal). Adicionalmente, os valores referentes às taxas metabólicas de neonatos machos e fêmeas lesados e controles não diferiram significativamente, seja em condição de normóxia normocápnica, hipercapnia ou hipóxia. Esses resultados sugerem que os neurônios catecolaminérgicos localizados no tronco encefálico exercem um papel inibitório tônico sobre ventilação em neonatos P7-8 e apresentam uma importante modulação excitatória na resposta ventilatória ao CO2 e hipóxia em ratas e ratos neonatos (P7-8) não anestesiados.

Fenilcetonúria (PKU) é um erro inato do metabolismo causado pela deficiência da atividade da enzima fenilalanina hidroxilase, levando ao acúmulo de fenilalanina e seus metabólitos no sangue e tecidos. A hiperfenilalaninemia (HPA) causa danos importantes no cérebro, provavelmente causados por aumento de estresse oxidativo e diminuição da disponibilidade dos outros aminoácidos grandes neutros (LNAA), entre outros mecanismos. Pacientes diagnosticados precocemente também estão sujeitos a estes desequilíbrios. O objetivo deste trabalho foi verificar em ratos: a) o efeito agudo do modelo de HPA na concentração de aminoácidos em plasma e cérebro total, b) o efeito do exercício regular em parâmetros de estresse oxidativo em cérebro total, conteúdo de catecolaminas em supra-renal e aspectos comportamentais na HPA crônica. Para o modelo agudo, os ratos foram divididos nos grupos HPA e Salina (SAL) (n=3). A HPA foi induzida através da administração subcutânea de alfa-metil-fenilalanina e fenilalanina, enquanto o grupo SAL recebeu salina. Os animais foram mortos 1 h após a injeção, no segundo dia de tratamento. Para o modelo crônico, os animais foram distribuídos no grupo Sedentário (Sed) ou Exercício (Exe), e subdivididos em SAL e HPA. Grupos HPA (n=16- 20) foram submetidos ao modelo durante 17 dias, enquanto os grupos SAL (n=16-20) receberam salina. Os grupos Exe realizaram duas semanas de exercício aeróbico com duração diária de 20 min. No 17º dia, 1 h após a injeção, os animais realizaram a primeira exposição ao teste de campo aberto e, 24 h depois, realizaram a segunda sessão. Após, os animais foram mortos e o cérebro total foi homogeneizado para determinação da lipoperoxidação, através do conteúdo de substâncias reativas ao ácido tiobarbitúrico (TBA-RS), e atividade das enzimas antioxidantes superóxido dismutase (SOD), catalase (CAT) e glutationa peroxidase (GPx). As glândulas supra-renais foram coletadas para análise de conteúdo de catecolaminas. O efeito agudo de HPA causou aumento de fenilalanina e diminuição de tirosina em plasma e cérebro, bem como diminuiu os níveis dos outros LNAA apenas no cérebro. Cronicamente, a HPA causou aumento de TBA-RS e SOD, e redução de CAT, GPx e conteúdo de catecolaminas. O exercício foi capaz de reverter todas as alterações encontradas no grupo HPA, exceto para a SOD. Quanto aos parâmetros comportamentais, a HPA causou diminuição na memória de habituação e o exercício regular preveniu esta alteração. Nenhuma alteração foi encontrada no grupo ExeSAL. Os ratos hiperfenilalaninêmicos foram mais responsivos aos benefícios produzidos pelo exercício regular. O treinamento físico parece ser uma estratégia interessante a ser estudada para a restauração do sistema antioxidante e de alterações comportamentais que ocorrem na PKU. / Phenylketonuria (PKU) is an inborn error of metabolism caused by deficiency of phenylalanine hydroxylase, resulting in accumulation of phenylalanine and its metabolites in blood and tissues. Hyperphenylalaninemia (HPA) causes serious damage in the brain probably due to increased oxidative stress and decreased availability of other large neutral amino acids (LNAA), among other mechanisms. Patients early diagnosed are also subject to these imbalances. The objective of this study was to evaluate: a) the effect of acute HPA model on the concentration of LNAA in plasma and total brain, b) the effect of regular exercise on parameters of oxidative stress in total brain, catecholamine content in suprarenal and behavioral aspects in a chronic HPA model. HPA was induced by subcutaneous administration of alpha-methylphenylalanine and phenylalanine, while SAL group received saline. For the acute model, rats were divided into groups Saline (SAL) and HPA (n = 3). Animals were killed 1 h after last injection, at the second day of treatment. For the chronic model, animals were divided into sedentary group (Sed) or exercise group (Exe), and subdivided into SAL (n=16-20) and HPA (n=16-20). Administration continued as long as 17 days. Exe groups performed two weeks of daily aerobic exercise lasting 20 min. At the 17th day, 1 h after injection, the animals performed the first exposure to open field task, and 24 h later, performed the second session. After that, animals were killed and the whole brain was homogenized to evaluate lipid peroxidation through the content of thiobarbituric acid reactive substances (TBA-RS), and activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Suprarenal glands were collected for catecholamine content analysis. Acute HPA increased phenylalanine and decreased tyrosine in plasma and brain as well as decreased levels of other LNAA in the brain. Chronically, HPA increased TBA-RS and SOD activity, and reduced CAT and GPx activities in the brain and reduced catecholamine content into suprarenal. Regular exercise was able to prevent all the alterations found in HPA group, except for SOD activity. Regarding the behavioral data, HPA caused a decrease of habituation memory and regular exercise prevented this change. Exercise per se (ExeSAL group) produced no changes. HPA rats were more responsive to the benefits produced by regular exercise. Physical training appears to be an interesting strategy to be studied for the restoration of the antioxidant system and the behavioral changes that occur in PKU.

Fenilcetonúria (PKU) é um erro inato do metabolismo causado pela deficiência da atividade da enzima fenilalanina hidroxilase, levando ao acúmulo de fenilalanina e seus metabólitos no sangue e tecidos. A hiperfenilalaninemia (HPA) causa danos importantes no cérebro, provavelmente causados por aumento de estresse oxidativo e diminuição da disponibilidade dos outros aminoácidos grandes neutros (LNAA), entre outros mecanismos. Pacientes diagnosticados precocemente também estão sujeitos a estes desequilíbrios. O objetivo deste trabalho foi verificar em ratos: a) o efeito agudo do modelo de HPA na concentração de aminoácidos em plasma e cérebro total, b) o efeito do exercício regular em parâmetros de estresse oxidativo em cérebro total, conteúdo de catecolaminas em supra-renal e aspectos comportamentais na HPA crônica. Para o modelo agudo, os ratos foram divididos nos grupos HPA e Salina (SAL) (n=3). A HPA foi induzida através da administração subcutânea de alfa-metil-fenilalanina e fenilalanina, enquanto o grupo SAL recebeu salina. Os animais foram mortos 1 h após a injeção, no segundo dia de tratamento. Para o modelo crônico, os animais foram distribuídos no grupo Sedentário (Sed) ou Exercício (Exe), e subdivididos em SAL e HPA. Grupos HPA (n=16- 20) foram submetidos ao modelo durante 17 dias, enquanto os grupos SAL (n=16-20) receberam salina. Os grupos Exe realizaram duas semanas de exercício aeróbico com duração diária de 20 min. No 17º dia, 1 h após a injeção, os animais realizaram a primeira exposição ao teste de campo aberto e, 24 h depois, realizaram a segunda sessão. Após, os animais foram mortos e o cérebro total foi homogeneizado para determinação da lipoperoxidação, através do conteúdo de substâncias reativas ao ácido tiobarbitúrico (TBA-RS), e atividade das enzimas antioxidantes superóxido dismutase (SOD), catalase (CAT) e glutationa peroxidase (GPx). As glândulas supra-renais foram coletadas para análise de conteúdo de catecolaminas. O efeito agudo de HPA causou aumento de fenilalanina e diminuição de tirosina em plasma e cérebro, bem como diminuiu os níveis dos outros LNAA apenas no cérebro. Cronicamente, a HPA causou aumento de TBA-RS e SOD, e redução de CAT, GPx e conteúdo de catecolaminas. O exercício foi capaz de reverter todas as alterações encontradas no grupo HPA, exceto para a SOD. Quanto aos parâmetros comportamentais, a HPA causou diminuição na memória de habituação e o exercício regular preveniu esta alteração. Nenhuma alteração foi encontrada no grupo ExeSAL. Os ratos hiperfenilalaninêmicos foram mais responsivos aos benefícios produzidos pelo exercício regular. O treinamento físico parece ser uma estratégia interessante a ser estudada para a restauração do sistema antioxidante e de alterações comportamentais que ocorrem na PKU. / Phenylketonuria (PKU) is an inborn error of metabolism caused by deficiency of phenylalanine hydroxylase, resulting in accumulation of phenylalanine and its metabolites in blood and tissues. Hyperphenylalaninemia (HPA) causes serious damage in the brain probably due to increased oxidative stress and decreased availability of other large neutral amino acids (LNAA), among other mechanisms. Patients early diagnosed are also subject to these imbalances. The objective of this study was to evaluate: a) the effect of acute HPA model on the concentration of LNAA in plasma and total brain, b) the effect of regular exercise on parameters of oxidative stress in total brain, catecholamine content in suprarenal and behavioral aspects in a chronic HPA model. HPA was induced by subcutaneous administration of alpha-methylphenylalanine and phenylalanine, while SAL group received saline. For the acute model, rats were divided into groups Saline (SAL) and HPA (n = 3). Animals were killed 1 h after last injection, at the second day of treatment. For the chronic model, animals were divided into sedentary group (Sed) or exercise group (Exe), and subdivided into SAL (n=16-20) and HPA (n=16-20). Administration continued as long as 17 days. Exe groups performed two weeks of daily aerobic exercise lasting 20 min. At the 17th day, 1 h after injection, the animals performed the first exposure to open field task, and 24 h later, performed the second session. After that, animals were killed and the whole brain was homogenized to evaluate lipid peroxidation through the content of thiobarbituric acid reactive substances (TBA-RS), and activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Suprarenal glands were collected for catecholamine content analysis. Acute HPA increased phenylalanine and decreased tyrosine in plasma and brain as well as decreased levels of other LNAA in the brain. Chronically, HPA increased TBA-RS and SOD activity, and reduced CAT and GPx activities in the brain and reduced catecholamine content into suprarenal. Regular exercise was able to prevent all the alterations found in HPA group, except for SOD activity. Regarding the behavioral data, HPA caused a decrease of habituation memory and regular exercise prevented this change. Exercise per se (ExeSAL group) produced no changes. HPA rats were more responsive to the benefits produced by regular exercise. Physical training appears to be an interesting strategy to be studied for the restoration of the antioxidant system and the behavioral changes that occur in PKU.

L'hémangiome capillaire infantile (IH) est une tumeur vasculaire bénigne courante. Les formes sévères sont traitées depuis 2008 par propranolol bien que le mécanisme d’action reste à ce jour inexpliqué. La 1ère partie de ma thèse est une étude de tolérance chez les enfants traités par propranolol dans le cadre de l'ATU française. L'analyse de cette base de données prospective de 922 enfants a permis de confirmer la bonne tolérance du propranolol dans la population pédiatrique, et de cibler les situations à risque de complication. La 2e partie est une étude de la signalisation adrénergique à partir des tissus d'IH opérés. Tout d'abord, nous avons identifié par immunofluorescence les cellules exprimant les récepteurs béta-adrénergiques ADRB1, 2 et 3 : Nous avons observé une forte expression d'ADRB2 sur le mastocyte, et de manière plus modérée d'ADRB1 et 2 sur les vaisseaux. Ce profil d'expression était retrouvé quel que soit le degré d'involution de la tumeur, et était également observé sur des tumeurs vasculaires contrôles qui elles ne répondent pas au propranolol. Nous avons donc envisagé une activation des récepteurs ADRB propre aux IH prolifératifs. Cette hypothèse a été confirmée par la mise en évidence, de l'expression d'enzymes de synthèse des catécholamines et de marqueurs neuroendocrines sur les péricytes des IH prolifératifs, sur tissus inclus en paraffine et également péricytes d’IH mis en culture. La surexpression de HIF1-a sur ces cellules a conduit à tester l’effet du propranolol en hypoxie. Nous n’avons pas mis en évidence d'effet sur la prolifération cellulaire, mais par contre une inhibition de la sécrétion de VEGF induite par l’hypoxie. / Capillary infantile haemangioma (IH) is a common benign vascular tumour of infants. Severe forms are treated since 2008 by propranolol. However, its dramatic efficacy remains until now unexplained.The first part of my thesis is a safety study of children treated with propranolol as part of the French Compassionate Use Program. The prospective vigilance database, including 922 children, confirmed the safety of propranolol among the paediatric patients, and highlighted the circumstances at risk of complications.The second part is a study of adrenergic signaling in IH tissues. First, we identified by immunofluorescence the expression of beta-adrenergic receptors on IH cells. We observed a strong expression of ADRB2 on mast cells, and a moderate ADRB1 and 2 expression on vessels. This expression pattern was the same regardless of the degree of involution of the tumour, and has also been observed on tumour vascular controls which did not respond to propranolol. We therefore hypothesized that ADRB receptors may be activated in proliferative IH. We indeed detected the expression of catecholamine synthesis enzymes and of neuro-endocrine markers on pericytes in paraffin-embedded tissues, and also in IH pericytes in culture. We also detected HIF1-alpha overexpression and therefore explored propranolol effect during hypoxia. Propranolol had no effect on cell proliferation, but reduced hypoxia-induced secretion of VEGF-A.

Although it is well-recognized that catecholamines are generally unable to penetrate the developed blood-brain barrier (BBB) to gain entry into brain, except at circumventricular sites where the BBB is absent or deficient, onto-genetic development of this barrier seems to have escaped systematic study. To explore BBB development, several approaches were used. In the first study rats were treated once on a specific day of postnatal ontogeny, as early as the day of birth, with the neurotoxin 6-hydroxydopa-mine (6-OHDA; 60 mg/kg), and then terminated in adulthood for regional analysis of endogenous norepinephrine (NE) content of brain. In another study, rats were treated once, on a specific day of postnatal ontogeny, with the BBB-perme-able neurotoxin 6-hydroxydopa (6-OHDOPA; 60 mg/kg) following pretreatment with the BBB-impermeable amino acid decarboxylase inhibitor carbidopa (100 mg/kg IP), then terminated in adulthood for regional analysis of endogenous NE content of brain. In the third study rats were treated once, on a specific day of postnatal ontogeny, with the analog [3H]metaraminol, and terminated 1 hour later for determination of regional distribution of tritium in brain. On the basis of [3H]metaraminol distribution and NE depletions after neurotoxin treatments, it is evident that the BBB in neocortex, striatum, cerebellum and other brain regions forms in stages over a period of at least 2 weeks from birth. Moreover, because the BBB consists of several element (physical-, ion-restrictive-, and enzymatic-barrier), the method employed will derive data mainly applicable to the targeted aspect of the barrier, which may or may not necessarily coincide with elements of the barrier that have a different rate of ontogenetic development. Accordingly, it is evident that some aspects of physical- and ion-restrictive elements of the BBB form within approximately the first week after birth in rat neocortex and striatum, while enzymatic elements of the BBB form more than than 2 week later. Regardless, the BBB forms at earlier times in forebrain vs hindbrain regions.

No description available.

No description available.

No description available.

The deer mouse (Peromyscus maniculatus) inhabits a broad altitudinal range from sea level to over 4300m, where they experience continuous hypoxia. Typically hypoxia activates the sympathetic nervous system; however this could become maladaptive in high-altitude residents if it is maintained over chronic periods. We hypothesized that high-altitude deer mice might have altered the physiology of adrenaomedullary chromaffin cells (AMC) in the adrenal gland to avoid chronic activation of the sympathetic response. Highland mice had lower plasma adrenaline levels compared to lowland populations of Peromyscus mice, both before and after acclimation to hypobaric hypoxia. This did not correspond to any apparent changes in AMC Ca2+-signalling dynamics. Instead a profound blunting of catecholamine storage was found in highland AMCs that appeared to underlie the reduction in adrenaline release / Thesis / Master of Science (MSc)

Das zeitgleiche Auftreten eines ischämischen Schlaganfalls sowie eines Takotsubo-Syndroms (TTS) scheint eine relevante, bisher nicht ausreichend verstandene klinische Konstellation zu sein. Die Pathologien können als über die Hirn-Herz-Achse gekoppelt verstanden werden, in die die Blut-Hirn-Schranke (BHS) als funktionale Komponente integriert ist. Das klinisch-neurologische Outcome dieses Patient:innen-Kollektivs scheint signifikant schlechter zu sein als nach solitärem ischämischen Insult. Es wurde hypothetisiert, dass die BHS in besonderem Maße kompromittiert sein könnte. Das vorwiegend weibliche, postmenopausale Patient:innenkollektiv präsentierte laborchemisch elevierte Katecholaminspiegel sowie Entzündungsparameter. Diese Konditionen wurden unter Sauerstoff-Glucose-Entzug (OGD) in vitro simuliert und resultierende Alterationen eines etablierten BHS-Modells aus murinen cEND-Zellen der cerebralen Mikrozirkulation untersucht. Die Evaluation der BHS-Integrität erfolgte anhand von spezifischen Junktionsproteinen sowie Integrinuntereinheiten. Alle Versuche wurden parallel unter Östrogen-Applikation (E2) durchgeführt, um die mögliche BHS-Protektion durch das weibliche Sexualhormon zu untersuchen. Die getrennte Applikation von Katecholaminen (KAT) sowie Entzündungsmediatoren (INF) führte gegenüber der simultanen Applikation zu einem geringeren BHS-Schaden. Dieser erschien zeitgebunden, wobei sich das Ausmaß gewissermaßen proportional zur Einwirkdauer verhielt. Auswirkungen von OGD sowie einer Reoxygenierung, im Sinne einer simulierten Reperfusion, potenzierten sich mit den Effekten von KAT/INF. Überwiegend kompromittierten OGD und KAT/INF die BHS-Integrität, wobei nach Reoxygenierung eine „Erholung“ oder ein „Reperfusionsschaden“ vorlag. Eine Protektion durch E2 war morphologisch nachweisbar, speziell gegenüber OGD, KAT/INF sowie einem „Reperfusionsschaden“. Auf Ebene der Gen- sowie Proteinexpression konnte dies nicht gezeigt werden. Die Homöostase des ZNS würde in vivo beeinträchtigt, Katecholamine sowie Entzündungsmediatoren könnten ungehindert das bereits durch die Ischämie geschädigte neuronale Gewebe erreichen. Insgesamt trägt diese Arbeit zu einem Verständnis der molekularen BHS-Veränderungen im Kontext des zeitgleichen Auftretens von TTS und einem ischämischem Insult bei. Es wurde eine experimentelle Grundlage geschaffen, um zukünftig pathogenetische Hintergründe weiter erforschen zu können. Darauf aufbauend könnten, nach weiterer in vitro- sowie in vivo-Forschung, klinische Therapiekonzepte optimiert werden. / The simultaneous occurrence of ischemic stroke and Takotsubo syndrome (TTS) seems to be a relevant clinical constellation that is not yet sufficiently understood. The pathologies can be understood as being linked via the brain-heart axis, into which the blood-brain barrier (BBB) is integrated as a functional component. The clinical and neurological outcome of these patients appears to be significantly worse than after a solitary ischemic insult. It has been hypothesized that the BBB may be compromised. The predominantly female, postmenopausal patients presented elevated catecholamine levels and inflammatory markers. These conditions were simulated in vitro under oxygen-glucose deprivation (OGD) condition. Resulting alterations were examined by using an established BBB model: cEND cells of the murine cerebral microcirculation. The BBB integrity was evaluated by investigating specific junction proteins and integrin subunits. All experiments were conducted parallel with estrogen application (E2) in order to investigate a possible BBB protection by the female sexhormone. The separate application of catecholamines (CAT) or inflammatory mediators (INF) led to less BBB damage compared to simultaneous application. This appeared to be time-bound being proportional to the duration of exposure. The effects of OGD and reoxygenation, in the sense of simulated reperfusion therapy, were potentiated by the effects of CAT/INF. Predominantly, OGD and KAT/INF compromised BBB integrity. “Recovery” or “reperfusion injury” occurred after reoxygenation. Protection by E2 was morphologically detectable, especially against OGD, CAT/INF and “reperfusion injury”. This could not be shown at the level of gene or protein expression, respectively. The homeostasis of the CNS would be impaired in vivo, catecholamines and inflammatory mediators would be able to reach the neuronal tissue that had already been damaged by ischemia. Overall, this work contributes to an understanding of the molecular changes in the BBB in the context of the simultaneous occurrence of TTS and ischemia. An experimental basis was created to enable further research into pathogenetic background. Based on this, clinical therapies could be optimized after further in vitro and in vivo research.