Bei der vorliegenden Arbeit handelt es sich um eine experimentelle Untersuchung zum Einfluss von zwei Ko-Transmittern des autonomen Nervensystems, Neuropeptid Y und dem Pituitary Adenylate Cyclase-activating Polypeptide (PACAP), auf den intrazellulären Kalziumspiegel und die Insulinsensitivität von 3T3-L1 Adipozyten. Mittels Polymerasekettenreaktion und Western Blot Analyse konnte die Expression des NPY-1 (Y1) Rezeptors als auch die der PACAP Rezeptoren PAC1 und VPAC2 nachgewiesen werden. Die Aktivierung des Y1 oder des PAC1 Rezeptors durch ihre Agonisten führte zur Erhöhung des intrazellulären Kalziumspiegels. Im Weiteren führte NPY nach Ko-Applikation mit Insulin zu einer abgeschwächten Insulinsensitivität der Adipozyten, da sowohl die insulin-stimulierte Translokation von Glukosetransporter 4 zur Zelloberfläche als auch die Glukoseaufnahme durch NPY abgeschwächt wurde. Dieser Effekt konnte als Y1 spezifisch beschrieben werden. Diese Ergebnisse gewähren somit neue Einblicke über den peptidergen Einfluss auf den Adipozytenstoffwechsel und erlauben Rückschlüsse über die Rolle des autonomen Nervensystems in der Entwicklung von Adipositas und Diabetes mellitus Typ 2.

nÃo hà / O gÃnero Aniba destaca-se por suas propriedades farmacolÃgicas, tais como ansiolÃtica, antidepressiva e vasorelaxante. Dos frutos de Aniba panurensis conhecida popularmente por âlouro amareloâ foi identificada uma pirona natural, a 6-[(E)-esteril-piran-2-ona]. O objetivo do presente trabalho foi verificar os efeitos neurofarmacolÃgicos da 6-[(E)-esteril-piran-2-ona (STY) obtida da Aniba parusinensi em camundongos atravÃs de testes comportamentais (atividade locomotora espontÃnea (ALE), rearing e grooming), testes de induÃÃo de convulsÃo quÃmica e dosagens neuroquÃmicas de aminoÃcidos (glutamato (GLU), aspartato (ASP), Ãcido γ-aminobutÃrico (GABA), glicina (GLI), taurina (TAU), histidina (HIS) em cÃrtex prÃ-frontal (CPF), hipocampo (HC) e corpo estriado (CE). Foram utilizados camundongos Swiss, machos, com peso mÃdio de 28 gramas. Os animais foram tratados com dose Ãnica de STY (1, 5, 10 ou 20 mg) por via intraperitoneal. Trinta minutos apÃs administraÃÃo os animais foram submetidos aos testes comportamentais e teste de convulsÃo induzidas por pentilenotetrazol (PTZ), estricnina, bicuculina e pilocarpina. Imediatamente apÃs os testes os animais foram sacrificados e as Ãreas cerebrais de interesse dissecadas para a dosagem da concentraÃÃo de aminoÃcidos atravÃs de HPLC (High Perfomance Liquid Chromatography). No teste de ALE a STY na dose de 10 ou 20 mg/kg aumentou atividade locomotora quando comparada ao grupo controle. No Labirinto em cruz elevada e o teste da placa perfurada, a STY em todas as doses comprovou seu efeito ansiolÃtico, pois aumentou todos os parÃmetros analisados. Na dosagem de aminoÃcidos neurotransmissores apÃs o teste de comportamento houve aumento nas concentraÃÃes dos aminoÃcidos inibitÃrios (GABA, GLI, TAU, HIS) e excitatÃrios (ASP, GLU) no CPF, HC e CE. ApÃs o prÃ-tratamento com STY os animais foram submetidos ao teste de induÃÃo de convulsÃo por PTZ (85 mg/kg) ou Bicuculina (12 mg/kg) foram observados aumentos na latÃncia de convulsÃo e morte, com animais sobreviventes na maior dose. No teste com a induÃÃo de convulsÃo por estricnina (20 mg/kg/ ocorreu o aumento na latÃncia de convulsÃo (STY-10: 50,42  7,20; STY-20: 65,99  3,22) e latÃncia de morte (STY-1: 20  1,72; STY-10: 19,17  1,87; STY-20: 23,83  1,55) em todos os animais prÃ-tratados com STY. Na induÃÃo de convulsÃo por pilocarpina ocorreu uma diminuiÃÃo da latÃncia de morte. ApÃs os testes os animais realizou-se a dosagem da concentraÃÃo dos aminoÃcidos (ASP, GLU, GLI, TAU e GABA). No CPF aumentaram ASP, TAU, GABA. Jà no HC o aumentou GLU, ASP, GLI e GABA. Na induÃÃo de convulsÃo com estricnina no CPF ocorreu o aumento no ASP, TAU, GABA, enquanto no HC aumentou ASP, GLI, TAU, GABA. No modelo de induÃÃo com bicuculina no CPF aumentou o ASP, GLU, GLI, TAU, GABA, enquanto no HC aumentou ASP, GLU, GLI, TAU, GABA. No modelo de induÃÃo com pilocarpina no CPF aumentou ASP, GABA, GLI, enquanto no HC aumentou apenas o ASP. Conclui-se que a STY apresenta um efeito ansiolÃtico nos testes de comportamento, efeito protetor nos teste de induÃÃo de convulsÃo por PTZ, estricnina e bicuculina. ApÃs o doseamento dos aminoÃcidos pode-se demonstrar a participaÃÃo dos sistemas glutamatÃrgico, GABAÃrgico e glinÃrgico. / The genus Aniba stands out for its pharmacological properties, such as anxiolytic, antidepressant and vasorelaxant. Of the Aniba panurensis fruits popularly known as ―louro amarelo‖ a natural pyrone, 6-[(E)-styryl-pyran-2-one], was identified. The objective of this study was to verify the neuropharmacological effects of 6-[(E)-styryl-pyran-2-one] (STY), obtained from Aniba panurensis, in mice through behavioral tests (spontaneous locomotor activity (SLA), rearing and grooming), tests of chemically induced seizures and neurochemical dosages of amino acids (glutamate (GLU), aspartate (ASP), γ-aminobutyric acid (GABA), glycine (GLY), taurine (Tau), histidine (HIS) in the prefrontal cortex (PFC), hippocampus (HC) and striatum (ST). We used Swiss mice, male, with an average weight of 28 grams. The animals were treated with a single dose of STY (1, 5, 10 or 20 mg) by intraperitoneal injection. Thirty minutes after administration, the animals were subjected to behavioral tests of chemically induced seizures by pentylenetetrazol (PTZ), strychnine, bicuculline and pilocarpine. Immediately after the tests, the animals were sacrificed and the brain areas of interest were dissected to estimate the amino acid concentration via HPLC (High Performance Liquid Chromatography). In the SLA test the STY at 10 or 20 mg/kg dose increased locomotor activity when compared to the control group. In the elevated plus-maze and hole-board tests, the STY in all doses proved its anxiolytic effect, because it increased all parameters analyzed. In the dosage of amino acid neurotransmitters after behavioral test there was an increase in inhibitory amino acid concentrations (GABA, GLY, TAU, HIS) and excitatory (ASP, GLU) in PFC, HC and ST. After pretreatment with STY, the animals were tested for seizures induced by PTZ (85 mg/kg) or Bicuculline (12 mg/kg), we observed an increase in the latency of seizures and death in surviving animals at the highest dose. During the strychnine-induced seizures test (20 mg/kg) there was an increase in seizure latency (STY-10: 50.42  7.20; STY-20: 65.99  3.22) and latency to death (STY-1: 20  1.72; STY-10: 19.17  1.87; STY-20: 23.83  1.55) in all animals pretreated with STY. In the pilocarpine-induced seizures there was a decrease in the latency to death. After testing the animals, we conducted the dosage of amino acid concentrations (ASP, GLU, GLY, TAU and GABA). PFC increased ASP, TAU and GABA. HC increased GLU, ASP, GLY and GABA. In the strychnine-induced seizure in PFC there was an increase in ASP, TAU and GABA, while the HC increased ASP, GLY, TAU and GABA. In the bicuculline-induced seizure in PFC there was an increase in ASP, GLU, GLY, TAU and GABA, while the HC increased ASP, GLU, GLY, TAU and GABA. In the pilocarpine-induced seizure in PFC there was an increase in ASP, GABA, GLY, while the HC increased only ASP. We concluded that STY presents an anxiolytic effect in behavioral tests and a protective effect in tests of induced seizures by PTZ, strychnine and bicuculline. After the dosage of the amino acids we can demonstrate the involvement of glutamatergic, GABAergic and glycinergic systems.

Cancer-induced bone pain is characterized by moderate to severe ongoing pain that commonly requires the use of opiates. Even when ongoing pain is well controlled, patients can suffer breakthrough pain (BTP), episodic severe pain that "breaks through" the medication. We developed a novel model of cancer-induced BTP using female rats with mammary adenocarcinoma cells sealed within the tibia. We demonstrated previously that rats with bone cancer learn to prefer a context paired with saphenous nerve block to elicit pain relief (i.e., conditioned place preference, CPP), revealing the presence of ongoing pain. Treatment with systemic morphine abolished CPP to saphenous nerve block, demonstrating control of ongoing pain. Here, we show that pairing BTP induced by experimenter-induced movement of the tumor-bearing hindlimb with a context produces conditioned place avoidance (CPA) in rats treated with morphine to control ongoing pain, consistent with clinical observation of BTP. Preventing movement-induced afferent input by saphenous nerve block before, but not after, hindlimb movement blocked movement-induced BTP. Ablation of isolectin B4 (IB4)-binding, but not TRPV1(+), sensory afferents eliminated movement-induced BTP, suggesting that input from IB4-binding fibers mediates BTP. Identification of potential molecular targets specific to this population of fibers may allow for the development of peripherally restricted analgesics that control BTP and improve quality of life in patients with skeletal metastases.

Bei der vorliegenden Arbeit handelt es sich um eine experimentelle Untersuchung zum Einfluss von zwei Ko-Transmittern des autonomen Nervensystems, Neuropeptid Y und dem Pituitary Adenylate Cyclase-activating Polypeptide (PACAP), auf den intrazellulären Kalziumspiegel und die Insulinsensitivität von 3T3-L1 Adipozyten. Mittels Polymerasekettenreaktion und Western Blot Analyse konnte die Expression des NPY-1 (Y1) Rezeptors als auch die der PACAP Rezeptoren PAC1 und VPAC2 nachgewiesen werden. Die Aktivierung des Y1 oder des PAC1 Rezeptors durch ihre Agonisten führte zur Erhöhung des intrazellulären Kalziumspiegels. Im Weiteren führte NPY nach Ko-Applikation mit Insulin zu einer abgeschwächten Insulinsensitivität der Adipozyten, da sowohl die insulin-stimulierte Translokation von Glukosetransporter 4 zur Zelloberfläche als auch die Glukoseaufnahme durch NPY abgeschwächt wurde. Dieser Effekt konnte als Y1 spezifisch beschrieben werden. Diese Ergebnisse gewähren somit neue Einblicke über den peptidergen Einfluss auf den Adipozytenstoffwechsel und erlauben Rückschlüsse über die Rolle des autonomen Nervensystems in der Entwicklung von Adipositas und Diabetes mellitus Typ 2.

Peptiderge Mediatoren sind neben ihrer Funktion bei der Aufrechterhaltung der körpereigenen Homöostase unter physiologischen Bedingungen auch bei der Regulation pathopysiologischer und pathobiochemischer Prozesse chronisch-entzündlicher Erkrankungen maßgeblich beteiligt. In der vorliegenden Arbeit wurde diese Rolle durch Untersuchung des Expressionsprofils peptiderger Mediatoren und ihrer Rezeptoren unter normalen Bedingungen charakterisiert und auf dieser Grundlage Veränderungen des Mediatorstoffwechsels bei entzündlichen Erkrankungen erfasst. Aufgrund der geringen Kenntnisse bezüglich der Rolle anti-inflammatorischer Mediatoren wurde dabei insbesondere die Expression und Genregulation des Mediators VIP und seiner Rezeptoren untersucht. Dabei wurden molekularbiologische Methoden verwandt, um definierte Rezeptoren für VIP und verwandte Mediatoren in den Atemwegen und der Haut zu identifizieren. Im Anschluss daran wurde anhand verschiedener entzündlicher Erkrankungen der oberen Atemwege nachgewiesen, dass sich das peptiderge Mediatorprofil krankheitsspezifisch ändert und diese Subgruppen-spezifischen Änderungen nicht als ein universelles Epiphänomen der Entzündungsreaktion zu sehen sind. Ebenso konnte die Veränderung der Genexpression von Rezeptoren für peptiderge Mediatoren untersucht werden, wobei am Beispiel der Hypoxie die Induktion eines in den Atemwegen exprimierten Rezeptors nachgewiesen wurde. Am Beispiel der atopischen Dermatitis konnte darüber hinaus bewiesen werden, dass die Expression von VIP-Rezeptoren im Rahmen einer allergischen Erkrankung vermindert sein kann. Letztlich wurden ebenfalls mit VIP interferierende Transduktionsmechanismen untersucht, wobei die genauen Interaktionen peptiderger Mediatoren mit diesen intrazellulären Molekülen im Rahmen entzündlicher Erkrankungen noch aufzuschlüsseln sind. Die Ergebnisse der vorliegenden kumulativen Arbeit weisen in ihrer Gesamtheit auf eine wesentliche Bedeutung neurogener Mediatoren für pathophysiologische Mechanismen allergisch-entzündlicher Erkrankungen der Atemwege und Haut hin und lassen zukünftige therapeutische Ansätze auf Basis neuro-immunmodulierender Mechanismen sinnvoll erscheinen. / Peptidergic mediators participate next to their physiological role for numerous aspects of systemic and local homeostasis also in the regulation of pathophysiological and pathobiochemical processes in chronic inflammatory diseases. In the present study this role was investigated by assessing the expression profiles of peptidergic mediators and their receptors under physiological conditions. Basing on these findings differences of the mediator expression in inflammatory diseases were examined. Due to the relatively little knowledge on the role of potentially anti-inflammatory mediators the expression and gene regulation of the mediator VIP und its receptors were analysed. In this respect molecular techniques were used to assess distinct receptors for VIP and related mediators in the airways and skin. IN a next Step inflammatory diseases of the upper respiratory tract were examined and it was shown that the peptidergic mediators profile changes in relation to the disease entity and that this disease subtype-specific change is not a universal epiphenomenon of the ongoing inflammation. Also, alterations in the gene expression of peptidergic mediator receptors were analysed. Using hypoxia as an example the gene induction of airway-expressed receptors was demonstrated in relation to this stimulus. In further studies involving atopic dermatitis tissues a down-regulation of VIP receptor expression was demonstrated for allergic inflammatory conditions. In a last step VIP interfering signal transduction mechanisms were examined and future studies need to be carried out to fully assess the regulation of these interactions in relation to chronic inflammatory processes. The present results demonstrate an important role of neurogenic mediators in the pathophysiology of allergic inflammatory diseases of the airways and the skin and point to a potential use of neuro-immunomodulation in the future therapy of these diseases.