Bisacurone gel ameliorated burn wounds in experimental rats via its anti-inflammatory, antioxidant, and angiogenic properties

Purpose: To investigate putative mechanism of wound healing for chitosan-based bisacurone gel against secondary burn wounds in rats. Methods: A second-degree burn wound with an open flame using mixed fuel (2 mL, 20 seconds) was induced in Sprague Dawley rats (male, 180-220 g, n = 15, each) followed by topical treatments with either vehicle control (white petroleum gel, 1%), silver sulfadiazine (1%) or bisacurone gel (2.5, 5, or 10%) for 20 days. Wound contraction rate and paw withdrawal threshold were monitored on various days. Oxidative stress (superoxide dismutase, glutathione, malondialdehyde, and nitric oxide), pro-inflammatory cytokines (tumour necrosis factor-alpha, interleukins by enzyme-linked immunosorbent assay), growth factors (transforming growth factor-ß, vascular endothelial growth factor C using real time polymerase chain reaction and Western blot assay) levels, and histology of wound skin were assessed at the end. Results: Bisacurone gel showed 98.72% drug release with a 420.90­442.70 cps viscosity. Bisacurone gel (5 and 10%) significantly (p < 0.05) improved wound contraction rate and paw withdrawal threshold. Bisacurone gel attenuated oxidative stress, pro-inflammatory cytokines, and water content. It also enhanced angiogenesis (hydroxyproline and growth factor) and granulation in wound tissue than vehicle control. Conclusions: These findings suggested that bisacurone gel can be a potential candidate to treat burn wounds via its anti-inflammatory, antioxidant, and angiogenic properties.

Cellular components and TGF-beta1 content of a closed Tube system for Platelet Rich Plasma acquisition in horse] / [Componentes celulares e TGF-beta1 do plasma rico em plaquetas obtido por sistema fechado em equinos]

Platelet-rich plasma (PRP) has been proposed as an agent to accelerate the healing process and stimulate the regenerative capacity of tissues due to its abundance of growth factors. A large variety of kits and protocols are available to obtain PRP by different cell-separation systems. However, the lack of standardization may lead to inconsistent results. The aim of this study was to characterize cellular composition, platelet parameters using the ADVIA 120 flow cytometer, and TGF-ß1 concentration from the PRP product obtained through a closed system, using simple centrifugation. Six clinically healthy horses were used in this study. The protocol in the closed system resulted in approximately 1.6-fold higher platelet and approximately 2.0-fold lower white blood cell concentrations in comparison with whole blood values. The evaluated system was efficient in concentrating platelets and in retrieving a small number of leukocytes, using a protocol of single centrifugation at low speed.

O plasma rico em plaquetas (PRP) tem sido proposto como um agente para acelerar o processo de cicatrização e estimular a capacidade regenerativa dos tecidos, devido à sua abundância de fatores de crescimento. Uma grande variedade de kits e de protocolos está disponível para obtenção de PRP, utilizando-se diferentes sistemas de separação de células. No entanto, a falta de padronização pode levar a resultados inconsistentes. O objetivo deste estudo foi caracterizar a composição celular, os parâmetros plaquetários pelo citômetro de fluxo ADVIA 120 e a concentração de TGF-ß1 do PRP obtido em sistema fechado, por centrifugação simples. Seis cavalos clinicamente saudáveis ​​foram usados ​​neste estudo. O protocolo no sistema fechado resultou em concentrações de plaquetas aproximadamente 1,6 vez maior e concentrações de leucócitos aproximadamente 2,0 vezes menores em comparação com os valores do sangue total. O sistema avaliado foi eficiente na concentração de plaquetas e na recuperação de um pequeno número de leucócitos, utilizando um protocolo de centrifugação única em baixa velocidade.

Antifibrotic preventive effect of polyethylene glycol (PEG) 3350 in methotrexateinduced hepatoxicity model

Purpose: Liver damage caused by drugs and other chemicals accounts for about 5% of all cases. Methotrexate (MTX), a folic acid analogue, is a first-line synthetic antimetabolite agent routinely used in the treatment of rheumatoid arthritis and other autoimmune and chronic inflammatory diseases. Polyethylene glycol (PEG) has antioxidant activity. In this study, we evaluated biochemically and histopathologically the antifibrotic effect of PEG 3350 administered intraperitoneally to prevent methotrexate-induced liver damage in rats. Methods: A total of 30 male rats including 10 rats was given no drugs (normal group), and 20 rats received single-dose 20 mg/kg MTXfor induced liver injury in this study. MTX was given to 20 rats, which were divided in two groups. Group 1 rats was given PEG30 mg/kg/day (Merck) intraperitoneally, and Group 2 rats % 0.9 NaCl saline 1 mL/kg/day intraperitoneally daily for two weeks. Results: Transforming growth factor beta (TGF-ß), plasma malondialdehyde (MDA), liver MDA, serum tumour necrosis factor alpha (TNF-α), alanine aminotransferase and plasma pentraxin-3 levels and, according to tissue histopathology, hepatocyte necrosis, fibrosis and cellular infiltration were significantly better in MTX+PEG group than in MTX+saline group. Conclusions: PEG 3350 is a hope for toxic hepatitis due to other causes, since liver damage occurs through oxidative stress and cell damage, similar to all toxic drugs.

Protective effect and mechanism of Shenkang injection on adenine-induced chronic renal failure in rats

Purpose: To investigate the protective effects of Shenkang injection (SKI) on adenine-induced chronic renal failure (CRF) in rat. Methods: Sprague Dawley rats were randomly divided into five groups: control, model, and SKI groups (5, 10, 20 mL/kg). Rats in model and SKI groups were treated with adenine i.g. at a dose of 150 mg/kg every day for 12 weeks to induce CRF. Twelve weeks later, SKI was administered to the rat i.p. for four weeks. The effects of SKI on kidney injury and fibrosis were detected. Results: SKI inhibited the elevation of the urine level of N-acetyl-b-D-glucosaminidase, kidney injury molecule-1, beta-2-microglobulin, urea protein in CRF rats. The serum levels of uric acid and serum creatinine increased and albumin decreased in the model group, which was prevented by SKI. SKI inhibited the release of inflammatory cytokines and increasing the activities of antioxidant enzymes in serum. SKI inhibited the expression of transforming growth factor-β1, vascular cell adhesion molecule 1, intercellular adhesion molecule 1, collagen I, collagen III, endothelin-1, laminin in kidney of CRF rats. Conclusions: SKI protected against adenine-induced kidney injury and fibrosis and exerted anti-inflammatory, and antioxidant effects in CRF rats.

Healing effect of andiroba-based emulsion in cutaneous wound healing via modulation of inflammation and transforming growth factor beta 3

Purpose:To evaluate the effects and mechanisms of andiroba-based emulsion (ABE) topical treatment on full-thickness cutaneous wounds in rats.Methods:The wounds were harvested on days 3, 7, 15, and 20 post-surgery. Wound contraction rate, quantitative immunohistochemistry [macrophages, myofibroblasts, capillaries, collagens (col) I and III, transforming growth factor β3β (TGFβ3)], and tensile strength were assessed.Results:Treated wounds were smaller, contracted earlier and had increased angiogenesis, fewer CD68+ and M2 macrophages on days 7 and 15, but higher on day 20. Myofibroblasts appeared on days 3 to 7 in untreated wounds and on days 7 to 15 in treated wounds. TGFβ3 levels were higher in the treated wounds, less dense collagen fibers, lower col I/III ratios and a higher tensile strength.Conclusion:These results demonstrate the important anti-inflammatory role of treatment and the associated modulation of macrophages, myofibroblasts, and TGFβ3 levels. Collagen fibers in the treated wounds were more organized and less dense, similar to unwounded skin, which likely contributed to the higher tensile strength.(AU)

Effects of Deletion of the Connexin 32 gene on pro and Anti-inflammatories Aspects in the Ethidium Bromide Toxic Demyelination Model

Gap junctions are cellular structures that allow transit of molecules between cells, allowing intercellular signaling and transportation. They are formed by proteins denominated connexins and represent key structures in highly complex and integrated tissues, such as the central nervous system (CNS). The present study evaluates the effects of connexin 32 (Cx32) deletion upon CNS inflammation and regeneration/repair after 1, 3, 7, 10 and 20 days after intracerebral injection of ethidium bromide in Cx32 Knock Out and normal mice. To accomplish so, Real Time PCR gene expression quantification was performed upon Tumour Necrosis Factor alpha (TNFα), Transforming Growth Factor beta 1 (TGFß1), Metalloproteinase 3 (MMP3), Metalloproteinase 9 (MMP9) and Tissue Inhibitor of Metalloproteinases 1 (TIMP1) genes. Results indicate varying differences in the expression pattern, including difference in expression of all evaluated genes in the 3 days post injection period, apex of the acute inflammation mechanisms. These results suggest that Cx32 may perform important functions on molecular, inflammatory and regenerative/repair signalling in the CNS.(AU)

Radiographic and densitometric aspects of experimental radial fractures of dogs treated with platelet-rich plasma / Aspectos radiográficos e densitométricos de fraturas experimentais do rádio de cães tratadas com plasma rico em plaquetas

The present article aimed to assess bone healing of experimental radial fractures, treated or not with autologous PRP (platelet-rich plasma), by means of radiographic and densitometric studies. Eleven dogs were randomly allocated in two experimental groups: control group (CG n=6) and PRP group (PRPG n=5). All dogs underwent radial osteotomy and osteosynthese (external skeletal fixation) of the right forelimb, in which a 2.0 mm defect was created. The gap was filled with PRP in the treated group (PRPG); while CG received no treatment. Radiographic and densitometric studies were performed immediately at the end the procedure and 14, 21, 28, 35, 45 and 60 days after the surgery (M0, M14, M21, M28, M35, M45 e M60). Statistical analysis was performed using analysis of variance with repeated measures (ANOVA) followed by Tukey test to compare means Friedman and Dunn post hoc tests to compare times within each group; and Mann-Whitney to compare groups at all times (p<0.05). Regarding time, radiographic scores and densitometric values changed significantly from moment 0 to moment 60 within the PRPG. The comparison of the two groups shows that radiographic scores changed significantly from M28 and densitometric values from M45 and M60. It can be concluded that under the conditions of this study; PRP can be used as an adjutant therapy to promote bone healing of experimental radial fractures (gap 2.0 mm) of dogs treated with external skeletal fixation.

O presente artigo teve como objetivo avaliar a cicatrização óssea de fraturas experimentais do radio de cães, tratadas ou não com o PRP autógeno, por meio de estudos radiográfico e densitométrico. Foram utilizados 11 cães, alocados aleatoriamente em dois grupos experimentais: o grupo controle (G-controle, n=6) e o grupo PRP (G-PRP, n=5). Todos os animais foram submetidos à osteotomia e osteossíntese (fixador esquelético externo) do rádio direito, gerando-se um gap de 2mm, que foi preenchido com PRP apenas no grupo G-PRP. Os estudos radiográficos e densitométricos foram realizados no pós-operatório imediato e aos 14, 21, 28, 35, 45 e 60 dias de pós-operatório (M0, M14, M21, M28, M35, M45 e M60). A análise estatística foi realizada por meio da Análise de Variância com Medidas repetidas (ANOVA) e teste de Tukey para comparação de médias; teste de Friedman seguido do teste de Dunn para comparar os momentos em cada grupo e o teste de Mann-whitney para comparar os grupos em cada momento (p<0,05). Referente à avaliação entre os momentos houve diferença significativa entre MO e M60 nas avaliações radiográficas e densitométricas dentro do grupo G-PRP. Na comparação entre os dois grupos houve diferença estatística na avaliação radiográfica a partir de M28; e na avaliação densitométrica nos momentos M45 e M60. Conclui-se, nas condições deste estudo, que o PRP pode ser utilizado como terapia adjuvante, pois promove cicatrização óssea precoce em fraturas experimentais (gap de 2,0 mm) do radio de cães tratadas com fixador esquelético externo.

Caracterizações celular e ultra-estrutural do concentrado de plaquetas em cães e gatos e avaliação do seu efeito na orteartrose em cães

Objetivou-se com este estudo: avaliar um método manual para obter concentrado autólogo de plaquetas (CAP) no cão e no gato para aplicação clínica, medir as concentrações do fator de crescimento beta transformador 1 (TGF-1) em cães e fator de crescimento derivado dasplaquetas tipo AB (PDGF-AB) em cães e gatos no plasma e nos concentrados autólogos de plaquetas (CAP) ativados com gluconato de cálcio ou cloreto de cálcio, medir as concentrações de TGF-1 em cães no plasma e nos CAP ativados com gluconato de cálcio ou batroxobina,medir as concentrações de TGF-1 e do fator de crescimento derivado das plaquetas tipo BB (PDGF-BB) em gatos no plasma e nos CAP ativados com gluconato de cálcio ou trombina bovina, avaliar por microscopia eletrônica de transmissão as características ultra-estruturais de coágulos sanguíneos de CAP ativados com gluconato de cálcio ou batroxobina, avaliar radiograficamente e em plataforma de força o efeito terapêutico do CAP no tratamento daosteoartrose (OA) adquirida no cão. Para a padronização do método do tubo para se obter CAP, utilizaram-se 12 cães e 12 gatos. Para avaliar a reprodutibilidade da metodologia e das características celulares dos CAP utilizaram-se 16 animais de cada espécie. Neste últimos animais o plasma derivado de centrifugação foi dividido em duas frações iguais, a fração A correspondeu aos primeiros 50% de plasma adjacente da interface sangue-plasma e a fração B correspondeu aos 50% do plasma restante no tubo. Nestes mesmos grupos foi feita a determinação dos fatores de crescimento. Na etapa de padronização o sangue foi colhido em frascos com EDTA para obtenção do plasma e o CAP foi obtido de sangue colhido num frasco com solução ACD-A. Foi medida a concentração de TGF-1 e PDGF-AB em cães e PDGF-AB no gato, no plasma e no sobrenadante de CAP ativado com gluconato ou cloreto de cálcio. Na etapa de reprodutibilidade o plasma e o CAP foram obtidos utilizando ACD-A. Foi medida a concentração de TGF-1 no plasma e no sobrenadante de CAP ativados com gluconato de cálcio ou batroxobina no cão, e a concentração de TGF-1 e PDGF-BB no plasma e no sobrenadante de CAP ativado com gluconato de cálcio ou trombina bovina no gato. Empregou-se um outro grupo de quatro cães e quatro gatos para avaliar as características ultra-estruturais de coágulossanguíneos obtidos de CAP ativado com gluconato de cálcio ou batroxobina. Foram feitas contagens celulares no sangue total e no CAP. Verificou-se em todas as amostras diferenças estatisticamente significativas (P<0,01) entre a quantidade de plaquetas, leucócitos,granulócitos, monócitos, mas não entre os valores de linfócitos, no sangue total e no CAP. A eficiência de concentração de plaquetas foi 22,20% a 46,34% nos cães e 24,75% a 35,39% nos gatos. A porcentagem de concentração de plaquetas nos CAP em relação ao sangue total foi 49,40% a 224,38% nos cães e 73,28% a 147,70% nos gatos. Em cães a concentração de TGF-1 foi estatisticamente (P<0,01) mais alta na fração A do CAP. No gato não se verificou diferençasignificativa (P>0,01) na concentração de TGF-1 e de PDGF-BB entre as frações A e B do CAP. Verificou-se nos cães diferença estatisticamente significativa (P<0,01) em relação à área de espaço intracelular e área de fibras de fibrina nos coágulos de CAP que foram ativados com gluconato de cálcio ou batroxobina. Em gatos os coágulos obtidos a partir de CAP ativado com batroxobina apresentaram diferença estatística (P<0,01), em termos de área das plaquetas, área de fibras de fibrina, relação eixo menor eixo maior, relação eixo maior eixo menor, nos coágulos que foram ativados com gluconato de cálcio ou batroxobina, os resultados demonstram maior grau de ativação das plaquetas ativadas com gluconato de cálcio. Para avaliar o efeito terapêutico do CAP, utilizou-se 10 cães de diferentes raças com ruptura de ligamento cruzado cranial (RLCCr) que foram submetidos a tratamento cirúrgico de substituição do ligamento por autoenxerto de fáscia lata guiado por videoartroscopia. Os cães foram divididos em dois grupos segundo o pós operatório: seis receberam três injeções intra-articulares de CAP (grupo I), iniciando imediatamente após a cirurgia e em intervalos de 15 dias, e o outro grupo constituído por quatro cães, receberam um agente modificador da osteoartrose (OA), via oral, diariamente, durante o tempo de estudo (grupo II). A avaliação radiográfica evidenciou, nos dois grupos de pacientes evolução discreta da OA. Verificou-se no grupo I melhor apoio na plataforma de força com evolução favorável e retorno do suporte no membro operado em todos os animais. Aos 90 dias o suporte no membro operado era igual ao do contralateral. No grupo II o apoio no membro operado evoluiu discretamente, ao longo dos 90 dias de avaliação, mantendo-se maior apoio no membro contralateral. Os resultados indicam que o método de centrifugação única permiteconcentrar plaquetas nestas duas espécies. O gluconato de cálcio é a substância indicada para ativação das plaquetas no cão e no gato, e a fração A do CAP no cão deve ser a empregada para fins terapêuticos, e no gato ambas frações podem ser usadas. Os resultados sugerem o possível potencial do concentrado autólogo de plaquetas (CAP) como alternativa biológica, versátil e econômica, como terapêutica no tratamento da OA

The objectives of this study were: to evaluate a manual method for obtaining autologous platelet concentrate (APC) in dog and cat for clinical purposes, measure the concentrations of transforming growth factor beta 1 (TGF-1) and platelet-derived growth factor type AB (PDGFAB) in dogs and cats, in plasma and autologous platelet concentrates (APC) activated with calcium gluconate or calcium chloride, measure the concentrations of TGF-1 in dogs, in plasma and APC activated with calcium gluconate or batroxobin, measuring the concentrations of TGF-1 and platelet derived growth factor type BB (PDGF-BB) in cats, in plasma and APCactivated with calcium gluconate or bovine thrombin, to evaluate by transmission electron microscopy ultrastructural characteristics of blood clots of APC activated with calcium gluconate or batroxobin, in dogs and cats, and too evaluate radiographic and plate force gait analysis, the therapeutic effect of APC in the treatment of osteoarthritis (OA) acquired in the dog. For standardization, we used 12 dogs and 12 cats. To evaluate the reproducibility of the methodology and cellular characteristics of APC, we used 16 animals of each species. These last dogs and cats, the plasma derived from the blood centrifugation was divided into two equalfractions, namely, PC-A and PC-B. Platelet concentrate-A (lower fraction) was considered as the first 50% plasma fraction near to the packed cell volume (PCV), and PC-B (upper fraction) represented the 50% remaining plasma. The same groups of animals were used for determining the concentration of growth factors. For standardization, to obtain the plasma blood was collected with EDTA and APC was obtained from blood collected with ACD-A solution. TGF- 1 and PDGF-AB concentrations after activation with gluconate or calcium chloride were measured in dogs and PDGF-AB in cats. To reproducibility, for blood collection was used vialswith ACD-A solution to obtain the plasma and APC. Was determined concentration of TGF-1 after activation with calcium gluconate or batroxobin in dogs; TGF-1 and PDGF-BB concentrations in cats, after activation with calcium gluconate or bovine thrombin. We used blood from another group of four dogs and four cats, for to evaluate the ultrastructuralcharacteristics of blood clots obtained of APC activated with calcium gluconate or batroxobin. Cell counts were performed in whole blood and the APC. It was found in all samples statistically significant (P <0.01) between the number of platelets, leukocytes, granulocytes, monocytes, but not between the values of lymphocytes in whole blood and the APC. Platelet efficiency concentration was 22.20% to 46.34% in dogs and 24.75% to 35.39% in cats. The percentage of platelet concentration in APC relative to the total blood was from 49.40% to 224.38% in dogs and 73.28% to 147.70% in cats. In dogs the concentration of TGF-1 wasstatically significantly higher (P <0.01) in fraction A of APC. In cats there was no significant difference (P> 0.01) in TGF-1 and PDGF-BB concentrations between the A and B fractions of APC. In dogs was found statistically significant difference (P<0.01) when compared intracellular space area and fiber area, in clots of APC that were activated with calciumgluconate or batroxobin. In cats clots obtained from APC activated with batroxobin show statistical difference (P <0.01) in area of platelets, area of fibrin fibers, ratio minor-major axes, ratio major-minor axes. We used 10 dogs, of different breeds with cranial cruciate ligament (CCL) rupture. These dogs were treated surgically by replacement of CCL with auto-graft of fascia lata, guided for video arthroscopy. They were divided into two groups according to postoperative period, six dogs received three intra-articular injections of autologous platelet concentrate (group I), starting immediately after surgery and at intervals of 15 days, and fourdogs received a modifying agent of OA daily, for all time of evaluation (group II). Radiographic evaluation showed, in both groups of patients, discrete evolution of OA. Group I was better in plate force gait analysis with favorable strength and returning the support member operated inall animals. At 90 days, the support of body mass on the operated limb was equal to that of contralateral limb. In-group II the support in the operated limb has evolved slightly, supporting the same body mass over 90th days. At 90 days, the support body mass of the contralateral limbwas greater than the affect limb. The results indicate that tube method allows for simple centrifugation concentrate platelets in dogs and cats. The calcium gluconate is the substance suitable for activation of APC in dogs and cats, and A fraction of APC in dogs must be used fortherapeutic purposes, in the cat both portions may be used. The results suggest the possible potential of APC as a biological alternative, versatile and economical, as a therapeutic option in the treatment of OA

Modulation of transforming growth factor beta2 (TGF-beta2) by inositol hexaphosphate in colon carcinogenesis in rats / Modulação do fator transformador de crescimento beta2 (TGFbeta2) pelo inositol hexafosfato na carcinogênese colônica em ratos

PURPOSE: To evaluate modulation in the expression of Transforming growth factor beta2 (TGF-beta2) in short-term colon carcinogenesis. METHODS: 64 male rats was used, comprising 4 groups of 16 animals each: group 1 received Inositol hexaphosphate (IP6) and azoxymethane (AOM); group 2, AOM alone; group 3, IP6 alone; group 4 was used as control. Groups 1 and 3 were given 1 percent IP6 in drinking water for 6 weeks. AOM was administered subcutaneously at weeks 3 and 4 of the experiment at 20 mg/kg of body weight each week. Immunohistochemical processing was performed with the use of anti-TGF-beta2 primary antibodies in right colon samples and quantitation of TGF-beta2 as percentage of expression, through computer-assisted image processing. RESULTS: mean values of TGF-beta2 expression were 9.0 ± 3.9 percent for group 4 (control), 12.7 ± 4.0 percent for group 3 (IP6), 19.3 ± 6.2 percent for group 2 (AOM), and 13.1 ± 5.3 percent for group 1 (IP6+AOM). The value of p was calculated as 0.0001 for a 5 percent or lower significance level. CONCLUSION: the experiment revealed a significant increase in TGF-beta2 expression in right colon with the administration of AOM, and a significant decrease in TGF-beta2 expression when IP6 was administered with AOM.(AU)

OBJETIVO: Avaliar a modulação da expressão do TGF-beta2 na carcinogênese colônica de curta duração em colon direito de ratos. Método: foram utilizados 64 ratos Wistar, machos divididos em 4 grupos de 16 animais. Grupo 1: recebeu Inositol hexafosfato (IP6) e azoximetano (AOM). Grupo 2 recebeu somente AOM. Grupo 3: recebeu somente IP6. Grupo 4: grupo de controle não recebeu nem IP6 nem AOM. O azoximetano (AOM) foi ministrado na dose 20mg/kg, por via subcutânea na 3ª e 4ª semanas do experimento. Foi realizada imunoistoquímica utilizando-se anticorpo primário TGFbeta2. Utilizou-se processamento de imagem computadorizada para quantificação da expressão do TGFbeta2. RESULTADOS: a média da expressão do TGFbeta2 foi de 9.0 ± 3.9 por cento para o grupo 4 (controle), 12.7 ± 4.0 por cento para o grupo 3 (IP6), 19.3 ± 6.2 por cento para o grupo 2 (AOM), e 13.1 ± 5.3 por cento para o grupo 1 (IP6+AOM). CONCLUSÃO: ocorreu aumento significante a da expressão de TGF-beta2 no cólon, com a administração de AOM, e uma diminuição significante na expressão de TGF-beta2 quando IP6 IP6 foi administrado com AOM.(AU)

Efeitos da deleção do gene para conexina 32 sobre aspectos pró e anti-inflamatórios no modelo de desmielinização tóxica do brometo de etídio / Effects of deletion of the connexin 32 gene on pro and antiinflammatories effects in the ethidium bromide toxic demyelination model

As junções comunicantes são estruturas celulares que permitem o trânsito de moléculas entre as células, desempenhando funções de sinalização e transporte intercelular. São formadas por proteínas denominadas conexinas e representam estruturas-chave no funcionamento de tecidos altamente complexos e integrados, como o sistema nervoso central (SNC). O presente estudo avalia os efeitos da deleção da conexina 32 (Cx32) na inflamação e regeneração/cicatrização do SNC após 1, 3, 7, 10 e 20 dias pós-injeção intracerebral de brometo de etídio em camundongos deletados para Cx32 e camundongos normais. Para tanto, quantificou-se a expressão dos genes para fator de necrose tumoral alfa (TNFα), fator de crescimento transformador beta 1 (TGFβ), metaloproteinase 3 (MMP3), metaloproteinase 9 (MMP9) e inibidor tecidual de metaloproteinases 1 (TIMP1), através de PCR em tempo real. Os resultados indicam variáveis diferenças no padrão de expressão, incluindo variação na expressão de todos os genes pesquisados no período de 3 dias pós-injeção, ápice dos mecanismos de inflamação aguda. Estes resultados sugerem que a conexina 32 pode desempenhar funções importantes na sinalização molecular do processo inflamatório e regenerativo/cicatricial do sistema nervoso central

Gap junctions are cellular structures that allow transit of molecules between cells, performing intercellular signaling and transportation. They are formed by proteins denominated connexins and represent key structures in highly complex and integrated tissues, such as the central nervous system (CNS). The present study evaluates the effects of connexin 32 (Cx32) deletion upon CNS inflammation and egeneration/cicatrization after 1, 3, 7, 10 and 20 days after intracerebral injection of ethidium bromide in Cx32 Knock Out and normal mice. To accomplish so, Real Time PCR gene expression quantification was performed uppon tumour necrosis factor alpha (TNFα), transforming growth factor beta 1 (TGFβ), metalloproteinase 3 (MMP3), metalloproteinase 9 (MMP9) and tissue inhibitor of metalloproteinases 1(TIMP1) genes. Results indicate variable differences in the expression pattern, including difference in expression of all evaluated genes in the 3 days after injection period, apex of the acute inflammation mechanisms. These results suggest that Cx32 may perform important functions on molecular inflammatory and regenerative/cicatrization signalling in the CNS

Effects of deletion of the connexin 32 gene on pro and anti-inflammatories aspects in the ethidium bromide toxic demyelination model

Gap junctions are cellular structures that allow transit of molecules between cells, allowing intercellular signaling and transportation. They are formed by proteins denominated connexins and represent key structures in highly complex and integrated tissues, such as the central nervous system (CNS). The present study evaluates the effects of connexin 32 (Cx32) deletion upon CNS inflammation and regeneration/repair after 1, 3, 7, 10 and 20 days after intracerebral injection of ethidium bromide in Cx32 Knock Out and normal mice. To accomplish so, Real Time PCR gene expression quantification was performed upon Tumour Necrosis Factor alpha (TNFa), Transforming Growth Factor beta 1 (TGFb1), Metalloproteinase 3 (MMP3), Metalloproteinase 9 (MMP9) and Tissue Inhibitor of Metalloproteinases 1 (TIMP1) genes. Results indicate varying differences in the expression pattern, including difference in expression of all evaluated genes in the 3 days post injection period, apex of the acute inflammation mechanisms. These results suggest that Cx32 may perform important functions on molecular, inflammatory and regenerative/repair signalling in the CNS.