RESUMO
Progesterone shows anti-inflammatory and promyelinating effects in mice with experimental autoimmune encephalomyelitis (EAE), a commonly used model for multiple sclerosis (MS). Because neurosteroids have been implicated as protective factors for MS and EAE, we analysed the expression of neurosteroidogenic enzymes in the compromised spinal cord of EAE mice. EAE was induced in female C57Bl6 mice, which were then killed on day 16 after induction. Progesterone was given by pellet implantation 1 week before EAE induction. Untreated EAE mice showed decreased mRNAs for the steroidogenic acute regulatory protein (Star), voltage-dependent anion channel (VDAC), cholesterol side-chain cleavage (P450scc), 5α-reductase, 3α-hydroxysteroid dehydrogenase (3α-HSOR) and aromatase, whereas changes of 3ß-hydroxysteroid dehydrogenase (3ß-HSD) were not significant. mRNA translocator protein (18 kDa) (TSPO) was elevated, concomitantly with a reactive microgliosis. EAE mice also showed abnormal mitochondrial ultrastructure in axons and neuronal bodies, as well as reduced expression of fission and fusion protein mRNAs. Progesterone pretreatment before EAE induction increased Star, VDAC, P450scc, 5α-reductase type I, 3α-HSOR and aromatase mRNAs and did not modify 3ß-HSD. TSPO mRNA was decreased, possibly as a result of reversal of microgliosis. Progesterone pretreatment also improved mitochondrial ultrastructure and increased fission/fusion protein mRNAs. These mitochondrial effects may be part of the progesterone recovery of neurosteroidogenesis. The enzymes 3ß-HSD, 3α-HSOR and 5α-reductase are also responsible for the formation of androgens. Because MS patients and EAE rodents show changes of central androgen levels, it is likely that, together with progestins and oestrogens, neuroandrogens afford neuroprotection for EAE and MS. The data reviewed suggest that enhanced synthesis of neurosteroids contributes in an auto/paracrine manner to reinforce the neuroprotective and anti-inflammatory effects of exogenous progesterone given to EAE mice.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Inflamação/tratamento farmacológico , Fármacos Neuroprotetores/uso terapêutico , Neurotransmissores/biossíntese , Progesterona/uso terapêutico , Animais , Encefalomielite Autoimune Experimental/metabolismo , Inflamação/metabolismo , Mitocôndrias/metabolismo , Fármacos Neuroprotetores/farmacologia , Progesterona/farmacologiaRESUMO
Wobbler mutant mice suffer from progressive motoneuron degeneration and glial cell reactivity in the spinal cord. To prevent development of these abnormalities, we employed Nestorone, a high-affinity progesterone receptor agonist endowed with neuroprotective, promyelinating and anti-inflammatory activities in experimental brain ischemia, preventing neuroinflammation and chemical degeneration. Five-month-old Wobbler mice (wr-/wr-) received s.c. injections of 200µg/day/mouse of Nestorone in vegetable oil or vehicle for 10days. Control NFR/NFR mice (background strain for Wobbler) received vehicle only. Vehicle-treated Wobblers showed typical spinal cord abnormalities, such as vacuolated motoneurons, decreased immunoreactive choline-acetyltransferase, decreased expression of glutamine synthase (GS), increased glial fibrillary acidic protein-positive (GFAP) astrogliosis and curved digits in forelimbs. These cell-specific abnormalities were normalized in Nestorone-treated Wobblers. In addition, vehicle-treated Wobblers showed Iba1+ microgliosis, high expression of the microglial marker CD11b mRNA and up-regulation of the proinflammatory markers TNFα and iNOS mRNAs. In Nestorone-treated Wobblers, Iba1+ microgliosis subsided, whereas CD11b, TNFα and iNOS mRNAs were down-regulated. NFκB mRNA was increased in Wobbler spinal cord and decreased by Nestorone, whereas expression of its inhibitor IκBα was increased in Nestorone-treated Wobblers compared to control mice and vehicle-treated Wobblers. In conclusion, our results showed that Nestorone restraining effects on proinflammatory mediators, microgliosis and astrogliosis may support neurons in their resistance against degenerative processes.
Assuntos
Anti-Inflamatórios/farmacologia , Doença dos Neurônios Motores/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Norprogesteronas/farmacologia , Receptores de Progesterona/agonistas , Medula Espinal/efeitos dos fármacos , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Astrócitos/patologia , Modelos Animais de Doenças , Gliose/tratamento farmacológico , Gliose/patologia , Gliose/fisiopatologia , Masculino , Camundongos Mutantes , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Doença dos Neurônios Motores/patologia , Doença dos Neurônios Motores/fisiopatologia , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Neuroimunomodulação/efeitos dos fármacos , Neuroimunomodulação/fisiologia , Receptores de Progesterona/metabolismo , Medula Espinal/metabolismo , Medula Espinal/patologia , Resultado do TratamentoRESUMO
Pressure ulcers are injuries to the skin and/or underlying tissues caused by prolonged high pressures on supporting body areas, they affect mainly people with poor mobility that have stayed in seating position for a long time. Reducing the amount and duration of pressure has been widely accepted for minimizing the risk of formation of pressure ulcers. Recently, dynamic cushions have been developed to relieve pressure on supporting areas; nevertheless, there is no sufficient information about the adequate characteristics of alternating sequences for pressure ulcers prevention. Therefore, the aim of this work is to explore three sequences of alternating movements designed for an air cell cushion by comparing pressure redistribution on supporting areas when applied on healthy volunteers. The purpose of these sequences is to redistribute the pressure over a larger contact area. To evaluate the effect of the alternating sequences, eight healthy volunteers were asked to sit on the air cell cushion, and to try the three alternating sequences for 12 minutes, 2 minutes on static mode and 10 minutes on alternating mode. A parameter for quantitative assessment of alternating sequences was proposed in this work by determining the coefficient of variation of interface pressure. Furthermore, the percentage of relative change of coefficient of variation was computed for evaluating performance of the alternating sequences comparing to the static mode. It was found that the three proposed strategies maintained values of interface pressure lower than previous work. Additionally, the relative change allowed to differentiate the effects of alternation of each sequence showing the second strategy as the most effective. The results are encouraging for further studies in subjects who require a wheelchair for mobility.
Las úlceras por presión son lesiones en la piel y tejidos subyacentes, causadas por presiones excesivas y prolongadas en las superficies de apoyo del cuerpo. Estas lesiones afectan principalmente a personas con poca movilidad física, como aquellas que permanecen sentados por largos periodos. Para disminuir el riesgo del padecimiento de estas lesiones, se ha recomendado como punto de partida reducir la magnitud y el tiempo de acción de las presiones en las zonas de apoyo. Se han desarrollado cojines dinámicos para sillas de ruedas, los cuales generan movimientos alternantes en las diferentes zonas de apoyo, producido por la inyección de aire, con el fin de disminuir las presiones en esas zonas. Sin embargo, no se han encontrado referencias de las características adecuadas de las secuencias de movimientos alternantes para prevenir la aparición de esas lesiones. El propósito de este trabajo es evaluar tres secuencias de movimientos alternantes diseñadas para un cojín de aire. La evaluación se realizó comparando la distribución de presiones en zonas de apoyo antes y durante la aplicación de estas secuencias alternantes en personas sanas. Las secuencias propuestas se aplican para el inflado y desinflado de celdas que forman el cojín y fueron diseñadas con el objetivo de distribuir las presiones en un área mayor de apoyo. La prueba se realizó en 8 sujetos sanos, con un tiempo de estudio de 12 minutos para cada secuencia diseñada; 2 minutos en modo estático y 10 minutos en modo alternante. Se propuso determinar el coeficiente de variación para evaluar de forma cuantitativa el efecto de las secuencias alternantes sobre la presión de interfaz. Además se calculó el porcentaje de variación relativa del coeficiente de variabilidad entre los modos basal (estático) y alternante como una herramienta para evaluar el desempeño de las secuencias propuestas en relación a la presión de interfaz. Se encontró que las tres estrategias mantuvieron presiones de interfaz por debajo de los valores reportados en trabajos previos. El porcentaje de variación relativa permitió diferenciar el efecto de la alternancia de cada una de las secuencias propuestas, mostrando la segunda estrategia como la más efectiva. Los resultados obtenidos son alentadores para continuar el estudio en sujetos que requieren una silla de ruedas para su movilidad.
RESUMO
Brazil has only one public genetic pool of Bombyx mori strains, which was established in 2005 at Universidade Estadual de Maringá, Maringá, Paraná State. This genetic bank has been maintained, and the strains have been characterized using genetic and morphological tools. The quantitative and qualitative traits, directly or indirectly related to productivity, were evaluated in 14 silkworm strains. In addition to biological and productivity analyses, DNA markers related to susceptibility to the B. mori nucleopolyhedrovirus (BmNPV) were analyzed. BmNPV is a major cause of production loss and is a serious problem for Paraná sericulture. The silkworm strains from diverse geographic origins were found to have different characteristics, including body weight, larval stage duration, cocoon weight, and other biological traits. In terms of productivity, the raw silk percentages were almost uniform, with an overall average of 16.28%. Overall, the Chinese strain C37 gave the best performance in many of the quantitative traits, and it surpassed the other strains in productivity traits. Therefore, it can be used as one of the strains that compose the elite germplasm for silkworm breeding programs. Additionally, genetic molecular markers were efficient in discriminating between B. mori strains that had been identified based on their geographical origin. We found that all Japanese strains produced a 400-bp molecular marker that has been associated with susceptibility to BmNPV.
Assuntos
Bombyx/genética , Bombyx/fisiologia , Marcadores Genéticos , Nucleopoliedrovírus/fisiologia , Animais , Bancos de Espécimes Biológicos , Bombyx/anatomia & histologia , Bombyx/virologia , Brasil , Cruzamento , Bases de Dados Genéticas , Larva/anatomia & histologia , Larva/genética , Larva/fisiologia , Característica Quantitativa Herdável , Seda , TêxteisRESUMO
Progesterone is a neuroprotective, promyelinating and anti-inflammatory factor for the nervous system. Here, we review the effects of progesterone in models of motoneurone degeneration and neuroinflammation. In neurodegeneration of the Wobbler mouse, a subset of spinal cord motoneurones showed increased activity of nitric oxide synthase (NOS), increased intramitochondrial NOS, decreased activity of respiratory chain complexes, and decreased activity and protein expression of Mn-superoxide dismutase type 2 (MnSOD2). Clinically, Wobblers suffered several degrees of motor impairment. Progesterone treatment restored the expression of neuronal markers, decreased the activity of NOS and enhanced complex I respiratory activity and MnSOD2. Long-term treatment with progesterone increased muscle strength, biceps weight and survival. Collectively, these data suggest that progesterone prevented neurodegeneration. To study the effects of progesterone in neuroinflammation, we employed mice with experimental autoimmune encephalomyelitis (EAE). EAE mice spinal cord showed increased mRNA levels of the inflammatory mediators tumour necrosis factor (TNF)α and its receptor TNFR1, the microglial marker CD11b, inducible NOS and the toll-like receptor 4. Progesterone pretreatment of EAE mice blocked the proinflammatory mediators, decreased Iba1+ microglial cells and attenuated clinical signs of EAE. Therefore, reactive glial cells became targets of progesterone anti-inflammatory effects. These results represent a starting point for testing the usefulness of neuroactive steroids in neurological disorders.
Assuntos
Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/tratamento farmacológico , Doenças Neurodegenerativas/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Progesterona/farmacologia , Progestinas/farmacologia , Animais , CamundongosRESUMO
In mice with experimental autoimmune encephalomyelitis (EAE) pretreatment with progesterone improves clinical signs and decreases the loss of myelin basic protein (MBP) and proteolipid protein (PLP) measured by immunohistochemistry and in situ hybridization. Presently, we analyzed if progesterone effects in the spinal cord of EAE mice involved the decreased transcription of local inflammatory mediators and the increased transcription of myelin proteins and myelin transcription factors. C57Bl/6 female mice were divided into controls, EAE and EAE receiving progesterone (100mg implant) 7 days before EAE induction. Tissues were collected on day 17 post-immunization. Real time PCR technology demonstrated that progesterone blocked the EAE-induced increase of the proinflammatory mediators tumor necrosis factor alpha (TNFα) and its receptor TNFR1, the microglial marker CD11b and toll-like receptor 4 (TLR4) mRNAs, and increased mRNA expression of PLP and MBP, the myelin transcription factors NKx2.2 and Olig1 and enhanced CC1+oligodendrocyte density respect of untreated EAE mice. Immunocytochemistry demonstrated decreased Iba1+microglial cells. Confocal microscopy demonstrated that TNFα colocalized with glial-fibrillary acidic protein+astrocytes and OX-42+microglial cells. Therefore, progesterone treatment improved the clinical signs of EAE, decreased inflammatory glial reactivity and increased myelination. Data suggest that progesterone neuroprotection involves the modulation of transcriptional events in the spinal cord of EAE mice.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/metabolismo , Mediadores da Inflamação/metabolismo , Bainha de Mielina/efeitos dos fármacos , Progesterona/farmacologia , Medula Espinal/metabolismo , Animais , Proteínas de Ligação ao Cálcio/biossíntese , Proteínas de Ligação ao Cálcio/genética , Regulação para Baixo/efeitos dos fármacos , Encefalomielite Autoimune Experimental/patologia , Feminino , Proteína Homeobox Nkx-2.2 , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Proteínas dos Microfilamentos/biossíntese , Proteínas dos Microfilamentos/genética , Microglia/metabolismo , Microscopia Confocal , Proteínas da Mielina/biossíntese , Bainha de Mielina/patologia , Oligodendroglia/efeitos dos fármacos , Oligodendroglia/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Medula Espinal/efeitos dos fármacos , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genéticaRESUMO
Previous results have shown a depletion of brain-derived neurotrophic factor (BDNF) mRNA in the degenerating motoneurons from clinically afflicted Wobbler mice, whereas progesterone treatment reverts this depletion. We now compared progesterone regulation of BDNF in motoneurons and oligodendrocytes of Wobbler mice at the progressive (EP, 1-3 months), symptomatic (SYM, 5-8 months old), and late stages (LS, 12-13 months). As controls we used NFR/NFR mice. Controls and Wobbler mice of different ages remained untreated or received a 20 mg progesterone pellet during 18 days. BDNF mRNA was determined in the ventral, intermediolateral, and dorsal gray matter by film autoradiography and in motoneurons using in situ hybridization. A depletion of BDNF mRNA already occurred at the EP stage of Wobblers, but progesterone was inactive at this period. In contrast, progesterone upregulated the low levels of BDNF mRNA in SYM Wobblers in the three gray matter regions analyzed. Progesterone also increased BDNF mRNA in LS Wobblers, according to grain counting procedures. BDNF protein analyzed by enzyme-linked immunosorbent assay (ELISA) in ventral horns or immunostaining of motoneurons was normal in steroid-naive SYM Wobblers. BDNF protein was decreased by progesterone, suggesting increased anterograde transport and/or release of neuronal BDNF. Wobbler mice also showed depletion of CC1-immunopositive oligodendrocytes, whereas progesterone treatment enhanced the density of BDNF+ and CC1+ oligodendrocytes in EP, SYM, and LS Wobblers. Our results suggest that BDNF could be involved in progesterone effects on motoneurons at the SYM and LS periods, whereas effects on oligodendrocytes occurred at all stages of the Wobbler disease. These steroid actions may be important to arrest the ongoing neurodegeneration.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Doença dos Neurônios Motores/metabolismo , Doença dos Neurônios Motores/patologia , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Progesterona/administração & dosagem , Fatores Etários , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Modelos Animais de Doenças , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica/genética , Camundongos , Camundongos Mutantes Neurológicos , Doença dos Neurônios Motores/tratamento farmacológico , Doença dos Neurônios Motores/genética , Mutação , Neuroglia/metabolismo , Neurônios/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Transporte Vesicular/genéticaRESUMO
Progesterone treatment of mice with experimental autoimmune encephalomyelitis has shown beneficial effects in the spinal cord according to enhanced clinical, myelin and neuronal-related parameters. In the present work, we report progesterone effects in a model of primary demyelination induced by the intraspinal injection of lysophospatidylcholine (LPC). C57Bl6 adult male mice remained steroid-untreated or received a single 100 mg progesterone implant, which increased circulating steroid levels to those of mouse pregnancy. Seven days afterwards mice received a single injection of 1% LPC into the dorsal funiculus of the spinal cord. A week after, anesthetized mice were perfused and paraffin embedded sections of the spinal cord stained for total myelin using Luxol Fast Blue (LFB) histochemistry, for myelin basic protein (MBP) immunohistochemistry and for determination of OX-42+ microglia/macrophages. Cryostat sections were also prepared and stained for oligodendrocyte precursors (NG2+ cells) and mature oligodendrocytes (CC1+ cells). A third batch of spinal cords was prepared for analysis of the microglial marker CD11b mRNA using qPCR. Results showed that progesterone pretreatment of LPC-injected mice decreased by 50% the area of demyelination, evaluated by either LFB staining or MBP immunostaining, increased the density of NG2+ cells and of mature, CC1+ oligodendrocytes and decreased the number of OX-42+ cells, respect of steroid-untreated LPC mice. CD11b mRNA was hyperexpressed in LPC-treated mice, but significantly reduced in LPC-mice receiving progesterone. These results indicated that progesterone antagonized LPC injury, an effect involving (a) increased myelination; (b) stimulation of oligodendrocyte precursors and mature oligodendrocytes, and (c) attenuation of the microglial/macrophage response. Thus, use of a focal demyelination model suggests that progesterone exerts promyelinating and anti-inflammatory effects at the spinal cord level.
Assuntos
Doenças Desmielinizantes/tratamento farmacológico , Microglia/efeitos dos fármacos , Progesterona/uso terapêutico , Medula Espinal/efeitos dos fármacos , Animais , Doenças Desmielinizantes/induzido quimicamente , Doenças Desmielinizantes/patologia , Modelos Animais de Doenças , Imuno-Histoquímica , Lisofosfatidilcolinas/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Medula Espinal/patologiaRESUMO
UNLABELLED: Negative prognostic factors in amyotrophic lateral sclerosis include advanced age, shorter time from disease onset to diagnosis, bulbar onset and rapid progression rate. OBJECTIVE: To compare progesterone (PROG) and cortisol serum levels in patients and controls and ascertain its relationship to prognostic factors and survival. METHODS: We assessed serum hormonal levels in 27 patients and 21 controls. RESULTS: Both hormones were 1.4-fold higher in patients. PROG showed a negative correlation with age, positive correlation with survival and positive trend with time to diagnosis. Increased PROG was observed in spinal onset and slow progression patients. No correlation was demonstrated with cortisol. CONCLUSION: Increased hormonal levels in patients are probably due to hypothalamic-pituitary-adrenal axis activation. Nevertheless, in this preliminary report only PROG correlated positively with factors predicting better prognosis and survival. We hypothesize endogenous PROG and cortisol may be engaged in differential roles, the former possibly involved in a neuroprotective response.