RESUMO
Inflammation and oxidative stress occurs in muscle of Duchenne muscular dystrophy (DMD). The relationship between a panel of biomarkers and the DMD outcome is necessary to indicate of disease progression and response to rehabilitation programs. The aim was to analyze the connective tissue of muscle of Mdx mice and immunoexpression of MMP-2, MMP-9, and 8-OHdG, which signalizes oxidative stress related to DNA damage. Biceps brachii of male C57BL/10 and C57BL/10-Dmdmdx mice was submitted to Hematoxylin-Eosin, Sirius red and immunohistochemistry (MMP-2, MMP-9 and 8-OHdG) analysis. Mdx showed focal lesions with intense inflammation and fibrosis related to immunoexpression of MMP-2 and MMP-9, proving the hypothesis that these MMPs are linked to muscular tissue degeneration, which can be regenerated by their inhibition, improving the treatment of DMD carriers. Histopathological findings related to centralized nuclei increase were related to higher 8-OHdG immunomarked nuclei in Mdx, which signalizes oxidative stress associated with DNA damage provoked by DMD. Such result shows that the evaluation of 8-OHdG during the evolution of the disease could be a method to evaluate DMD disease progression.
Assuntos
8-Hidroxi-2'-Desoxiguanosina/biossíntese , Regulação Enzimológica da Expressão Gênica , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Distrofia Muscular de Duchenne , Animais , Fibrose , Masculino , Camundongos , Camundongos Endogâmicos mdx , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/patologiaRESUMO
OBJECTIVE: To characterize the patterns of cell differentiation, proliferation, and tissue invasion in eutopic and ectopic endometrium of rabbits with induced endometriotic lesions via a well- known experimental model, 4 and 8 weeks after the endometrial implantation procedure. METHODS: Twenty-nine female New Zealand rabbits underwent laparotomy for endometriosis induction through the resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of animals (one with 14 animals, and the other with15) were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised along with the opposite uterine horn for endometrial gland and stroma determination. Immunohistochemical reactions were performed in eutopic and ectopic endometrial tissues for analysis of the following markers: metalloprotease (MMP-9) and tissue inhibitor of metalloprotease (TIMP-2), which are involved in the invasive capacity of the endometrial tissue; and metallothionein (MT) and p63, which are involved in cell differentiation and proliferation. RESULTS: The intensity of the immunostaining for MMP9, TIMP-2, MT, and p63 was higher in ectopic endometria than in eutopic endometria. However, when the ectopic lesions were compared at 4 and 8 weeks, no significant difference was observed, with the exception of the marker p63, which was more evident after 8 weeks of evolution of the ectopic endometrial tissue. CONCLUSION: Ectopic endometrial lesions seem to express greater power for cell differentiation and tissue invasion, compared with eutopic endometria, demonstrating a potentially invasive, progressive, and heterogeneous presentation of endometriosis.
OBJETIVO: Caracterizar o padrão de diferenciação celular, proliferação e invasão tecidual em endométrio eutópico e ectópico de coelhas com lesões de endometriose induzidas por um modelo experimental 4 e 8 semanas após o procedimento de implantação endometrial. MéTODOS: Vinte e nove coelhas fêmeas Nova Zelândia foram submetidas a laparotomia para indução de endometriose através da ressecção de um dos cornos uterinos, isolamento do endométrio e fixação do tecido no peritônio pélvico. Dois grupos de animais (14 animais em um grupo e 15 animais no outro) foram sacrificados 4 e 8 semanas após a indução da endometriose. A lesão foi excisada junto com o corno uterino contralateral para determinação da presença de glândulas e de estroma endometrial. Reações de imunohistoquímica foram realizadas no tecido endometrial eutópico e ectópico para análise dos seguintes marcadores: metaloprotease (MMP9) e inibidor tecidual da metaloprotease 2 (TIMP-2), os quais estão envolvidos na capacidade de invasão do tecido endometrial; e metalotioneina (MT) e p63, os quais estão envolvidos na diferenciação e proliferação celular. RESULTADOS: A intensidade da imunomarcação para MMP9, TIMP-2, MT e p63 foi mais alta nos endométrios ectópicos do que nos endométrios eutópicos. Contudo, quando as lesões foram comparadas entre 4 e 8 semanas, nenhuma diferença foi observada, com exceção do marcador p63, o qual foi mais evidente depois de 8 semanas de evolução do tecido endometrial ectópico. CONCLUSãO: Lesões endometriais ectópicas parecem expressar maior poder de diferenciação celular e de invasão tecidual comparadas com endométrios eutópicos, demonstrando o potencial de invasão, de progressão e de apresentação heterogênea da endometriose.
Assuntos
Coristoma/metabolismo , Endometriose/metabolismo , Endométrio/metabolismo , Metaloproteinase 9 da Matriz/biossíntese , Proteínas de Membrana/biossíntese , Metalotioneína/biossíntese , Inibidor Tecidual de Metaloproteinase-2/biossíntese , Animais , Diferenciação Celular , Proliferação de Células , Coristoma/patologia , Modelos Animais de Doenças , Endometriose/patologia , Endométrio/química , Endométrio/patologia , Feminino , Metaloproteinase 9 da Matriz/análise , Proteínas de Membrana/análise , Metalotioneína/análise , Coelhos , Inibidor Tecidual de Metaloproteinase-2/análiseRESUMO
Abstract Objective To characterize the patterns of cell differentiation, proliferation, and tissue invasion in eutopic and ectopic endometrium of rabbits with induced endometriotic lesions via a well- known experimental model, 4 and 8 weeks after the endometrial implantation procedure. Methods Twenty-nine female New Zealand rabbits underwent laparotomy for endometriosis induction through the resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of animals (one with 14 animals, and the other with15) were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised along with the opposite uterine horn for endometrial gland and stroma determination. Immunohistochemical reactions were performed in eutopic and ectopic endometrial tissues for analysis of the following markers: metalloprotease (MMP-9) and tissue inhibitor of metalloprotease (TIMP-2), which are involved in the invasive capacity of the endometrial tissue; and metallothionein (MT) and p63, which are involved in cell differentiation and proliferation. Results The intensity of the immunostaining for MMP9, TIMP-2, MT, and p63 was higher in ectopic endometria than in eutopic endometria. However, when the ectopic lesions were compared at 4 and 8 weeks, no significant difference was observed, with the exception of the marker p63, which was more evident after 8 weeks of evolution of the ectopic endometrial tissue. Conclusion Ectopic endometrial lesions seem to express greater power for cell differentiation and tissue invasion, compared with eutopic endometria, demonstrating a potentially invasive, progressive, and heterogeneous presentation of endometriosis.
Resumo Objetivo Caracterizar o padrão de diferenciação celular, proliferação e invasão tecidual em endométrio eutópico e ectópico de coelhas com lesões de endometriose induzidas por um modelo experimental 4 e 8 semanas após o procedimento de implantação endometrial. Métodos Vinte e nove coelhas fêmeas Nova Zelândia foram submetidas a laparotomia para indução de endometriose através da ressecção de um dos cornos uterinos, isolamento do endométrio e fixação do tecido no peritônio pélvico. Dois grupos de animais (14 animais em um grupo e 15 animais no outro) foram sacrificados 4 e 8 semanas após a indução da endometriose. A lesão foi excisada junto com o corno uterino contralateral para determinação da presença de glândulas e de estroma endometrial. Reações de imunohistoquímica foram realizadas no tecido endometrial eutópico e ectópico para análise dos seguintes marcadores: metaloprotease (MMP9) e inibidor tecidual da metaloprotease 2 (TIMP-2), os quais estão envolvidos na capacidade de invasão do tecido endometrial; e metalotioneina (MT) e p63, os quais estão envolvidos na diferenciação e proliferação celular. Resultados A intensidade da imunomarcação para MMP9, TIMP-2, MT e p63 foi mais alta nos endométrios ectópicos do que nos endométrios eutópicos. Contudo, quando as lesões foram comparadas entre 4 e 8 semanas, nenhuma diferença foi observada, com exceção do marcador p63, o qual foi mais evidente depois de 8 semanas de evolução do tecido endometrial ectópico. Conclusão Lesões endometriais ectópicas parecem expressar maior poder de diferenciação celular e de invasão tecidual comparadas com endométrios eutópicos, demonstrando o potencial de invasão, de progressão e de apresentação heterogênea da endometriose.
Assuntos
Animais , Feminino , Coristoma/metabolismo , Inibidor Tecidual de Metaloproteinase-2/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Endometriose/metabolismo , Endométrio/metabolismo , Proteínas de Membrana/biossíntese , Metalotioneína/biossíntese , Coelhos , Diferenciação Celular , Coristoma/patologia , Inibidor Tecidual de Metaloproteinase-2/análise , Metaloproteinase 9 da Matriz/análise , Proliferação de Células , Modelos Animais de Doenças , Endometriose/patologia , Endométrio/patologia , Endométrio/química , Proteínas de Membrana/análise , Metalotioneína/análiseRESUMO
Astrocytomas represent the majority of cerebral gliomas. Studies show that the anti-inflammatory protein Annexin-A1 (ANXA1) is associated with the tumor invasion process and that its actions can be mediated by the receptor for formylated peptides (FPR). Therefore, we evaluated the expression of ANXA1, the receptor FPR2 and matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) in brain astrocytomas. Detection of proteins was performed in sections of diffuse astrocytomas (grade II), anaplastic astrocytomas (grade III) and glioblastomas (GBM, grade IV) and quantifications were made by densitometry. Our analyses showed increased expression of ANXA1 in astrocytomas of all grades, but especially in GBM. The expression of FPR2 is similar to that found for ANXA1, being higher in GBM. Immunostaining for MMPs is also stronger as the degree of malignancy increases, especially with respect to MMP-9. The positive correlation between ANXA1/FPR2 and ANXA1/MMP-9 was observed in all tumors studied. The data indicate the possible action of ANXA1 and FPR2 on the development and progression of astrocytomas, related to increased expression of MMP-9. Thereby, ANXA1 and FPR2 are involved in the biology and malignancy of diffuse astrocytic tumors.
Assuntos
Anexina A1/biossíntese , Astrocitoma/patologia , Biomarcadores Tumorais/biossíntese , Neoplasias Encefálicas/patologia , Receptores de Formil Peptídeo/biossíntese , Receptores de Lipoxinas/biossíntese , Adulto , Idoso , Feminino , Humanos , Masculino , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Lymphangioleiomyomatosis (LAM) is a low-grade neoplasm characterized by the pulmonary infiltration of smooth muscle-like cells (LAM cells) and cystic destruction. Patients usually present with airway obstruction in pulmonary function tests (PFTs). Previous studies have shown correlations among histological parameters, lung function abnormalities and prognosis in LAM. We investigated the lung tissue expression of proteins related to the mTOR pathway, angiogenesis and enzymatic activity and its correlation with functional parameters in LAM patients. METHODS: We analyzed morphological and functional parameters of thirty-three patients. Two groups of disease severity were identified according to FEV1 values. Lung tissue from open biopsies or lung transplants was immunostained for SMA, HMB-45, mTOR, VEGF-D, MMP-9 and D2-40. Density of cysts, density of nodules and protein expression were measured by image analysis and correlated with PFT parameters. RESULTS: There was no difference in the expression of D2-40 between the more severe and the less severe groups. All other immunohistological parameters showed significantly higher values in the more severe group (p ≤ 0.002). The expression of VEGF-D, MMP-9 and mTOR in LAM cells was associated with the density of both cysts and nodules. The density of cysts and nodules as well as the expression of MMP-9 and VEGF-D were associated with the impairment of PFT parameters. CONCLUSIONS: Severe LAM represents an active phase of the disease with high expression of VEGF-D, mTOR, and MMP-9, as well as LAM cell infiltration. Our findings suggest that the tissue expression levels of VEGF-D and MMP-9 are important parameters associated with the loss of pulmonary function and could be considered as potential severity markers in open lung biopsies of LAM patients.
Assuntos
Biomarcadores Tumorais/biossíntese , Neoplasias Pulmonares/metabolismo , Linfangioleiomiomatose/metabolismo , Metaloproteinase 9 da Matriz/biossíntese , Serina-Treonina Quinases TOR/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Adulto , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/patologia , Linfangioleiomiomatose/patologia , Metaloproteinase 9 da Matriz/análise , Pessoa de Meia-Idade , Estudos Retrospectivos , Serina-Treonina Quinases TOR/análise , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
The skin is the first organ to be infected by the parasite in canine visceral leishmaniasis. The enzyme matrix metalloproteinase (MMP) acts towards degradation of the extracellular matrix (ECM) and modulation of the inflammatory response against many kinds of injuries. The aims of this study were to evaluate the expression of MMP-2 and MMP-9 through immunohistochemistry and zymography on the skin (muzzle, ears, and abdomen) of dogs that were naturally infected by Leishmania spp. and to compare these results with immunodetection of the parasite and with alterations to the dermal ECM. Picrosirius red staining was used to differentiate collagen types I and III in three regions of the skin. The parasite load, intensity of inflammation, and production of MMP-2 (latent) and MMP-9 (active and latent) were higher in the ear and muzzle regions. MMP-9 (active) predominated in the infected group of dogs and its production was significantly different to that of the control group. Macrophages, lymphocytes, and plasma cells predominated in the dermal inflammation and formed granulomas in association with degradation of mature collagen (type I) and with discrete deposition of young collagen (type III). This dermal change was more pronounced in dogs with high parasite load in the skin. Therefore, it was concluded that the greater parasite load and intensity of inflammation in the skin led consequently to increased degradation of mature collagen, caused by increased production of MMPs, particularly active MMP-9, in dogs with visceral leishmaniasis. This host response profile possibly favors systemic dissemination of the parasite.
Assuntos
Doenças do Cão/patologia , Leishmaniose Visceral/veterinária , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Pele/patologia , Abdome/parasitologia , Abdome/patologia , Animais , Colágeno Tipo I/metabolismo , Colágeno Tipo III/metabolismo , Doenças do Cão/parasitologia , Cães , Orelha/parasitologia , Orelha/patologia , Matriz Extracelular/metabolismo , Inflamação/imunologia , Inflamação/parasitologia , Leishmania infantum/imunologia , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/patologia , Linfócitos/imunologia , Macrófagos/imunologia , Boca/parasitologia , Boca/patologia , Nariz/parasitologia , Nariz/patologia , Carga Parasitária , Plasmócitos/imunologia , Pele/parasitologiaRESUMO
6-Gingerol is the major active constituent of ginger. In the current study, we aimed to investigate the mechanisms underlying the effects of 6-Gingerol on hair growth. Mice were randomly divided into five groups; after hair depilation (day 0), mice were treated with saline, or different concentrations of 6-Gingerol for 11 days. The histomorphological characteristics of the growing hair follicles were examined after hematoxylin and eosin staining. The results indicated that 6-Gingerol significantly suppressed hair growth compared with that in the control group. And choose the concentration of 6-Gingerol at 1 mg/mL to treated with mice. Moreover, 6-Gingerol (1 mg/mL) significantly reduced hair re-growth ratio, hair follicle number, and hair follicle length, which were associated with increased expression of MMP2 and MMP9. Furthermore, the growth factors, such as EGF, KGF, VEGF, IGF-1 and TGF-ß participate in the hair follicle cycle regulation and regulate hair growth. We then measured the concentrations of them using ELISA assays, and the results showed that 6-Gingerol decreased EGF, KGF, VEGF, and IGF-1 concentrations, and increased TGF-ß concentration. Thus, this study showed that 6-Gingerol might act as a hair growth suppressive drug via induction of MMP2 and MMP9 expression, which could interfere with the hair cycle.
Assuntos
Catecóis/farmacologia , Álcoois Graxos/farmacologia , Folículo Piloso/efeitos dos fármacos , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Extratos Vegetais/farmacologia , Animais , Indução Enzimática , Feminino , Fator 7 de Crescimento de Fibroblastos/biossíntese , Folículo Piloso/patologia , Fator de Crescimento Insulin-Like I/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Distribuição Aleatória , Fator de Crescimento Transformador beta/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossínteseRESUMO
ABSTRACT 6-Gingerol is the major active constituent of ginger. In the current study, we aimed to investigate the mechanisms underlying the effects of 6-Gingerol on hair growth. Mice were randomly divided into five groups; after hair depilation (day 0), mice were treated with saline, or different concentrations of 6-Gingerol for 11 days. The histomorphological characteristics of the growing hair follicles were examined after hematoxylin and eosin staining. The results indicated that 6-Gingerol significantly suppressed hair growth compared with that in the control group. And choose the concentration of 6-Gingerol at 1 mg/mL to treated with mice. Moreover, 6-Gingerol (1 mg/mL) significantly reduced hair re-growth ratio, hair follicle number, and hair follicle length, which were associated with increased expression of MMP2 and MMP9. Furthermore, the growth factors, such as EGF, KGF, VEGF, IGF-1 and TGF-β participate in the hair follicle cycle regulation and regulate hair growth. We then measured the concentrations of them using ELISA assays, and the results showed that 6-Gingerol decreased EGF, KGF, VEGF, and IGF-1 concentrations, and increased TGF-β concentration. Thus, this study showed that 6-Gingerol might act as a hair growth suppressive drug via induction of MMP2 and MMP9 expression, which could interfere with the hair cycle.
Assuntos
Animais , Masculino , Feminino , Coelhos , Extratos Vegetais/farmacologia , Catecóis/farmacologia , Folículo Piloso/efeitos dos fármacos , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Álcoois Graxos/farmacologia , Fator de Crescimento Insulin-Like I/biossíntese , Distribuição Aleatória , Indução Enzimática , Fator de Crescimento Transformador beta/biossíntese , Folículo Piloso/patologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator 7 de Crescimento de Fibroblastos/biossíntese , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVE: to evaluate the efficacy of three immunohistochemical markers involved in the wound healing process. METHODS: experimental study of 40 Wistar rats of the markers metalloproteinases and matrix metalloproteinase 9 (MMP-9), beta transforming growth factor (TGF-ß) and myofibroblasts and smooth muscle actin alpha (α-MLA) markers, studied from fragments of surgical scar of abdominal incision involving skin, aponeurosis and peritoneum. The animals were divided into four subgroups of ten according to the day of death, scheduled in three, seven, 14 and 21 days. RESULTS: MMP-9 expression showed a progressive increase of its concentration, more evident from 7th to 14th days, varying the tissue immunoexpression between 2.65% and 11.50% . TGF- ß showed expression at high level on the 3rd day, fell in the 7th, rising again in the 14th, with a small decrease in the 21st day, varying the tissue immunoexpression between 0.03% and 2.92%. The α-AML presented levels with little variation and a slight increase, varying the tissue immunoexpression between 0.88% and 3.23%. CONCLUSION: MMP-9 presented as the best marker, followed by TGF-ß. However, α-AML was not a good indicator of the evolution of tissue repair.
OBJETIVO: avaliar a eficácia de três marcadores imunoistoquímicos envolvidos no processo de cicatrização de ferida cirúrgica. MÉTODOS: estudo experimental em 40 ratos da raça Wistar, dos marcadores metaloproteinases e metaloproteinase da matriz 9 (MMP-9), fator de transformação do crescimento beta (TGF-ß) e miofibroblasto e alfa actina de músculo liso (α-AML), estudados a partir de fragmentos de cicatriz cirúrgica de incisão abdominal envolvendo pele, aponeurose e peritônio. Os animais foram distribuídos em quatro subgrupos de dez de acordo com o dia da morte, programada em três, sete, 14 e 21 dias. RESULTADOS: na expressão da MMP-9 ocorreu aumento progressivo de sua concentração, mais evidente do 7º ao 14º dias variando a imuno-expressão tecidual entre 2,65% e 11,50%.TGF- ß mostrou expressão em nível alto no 3º dia, caiu no 7º, voltando a subir no 14º, com pequena queda no 21º dia variando a imuno-expressão tecidual entre 0,03% e 2,92%. A α-AML apresentou níveis com pouca variação e discreto aumento variando a imuno-expressão tecidual entre 0,88% e 3,23%. CONCLUSÃO: a MMP-9 se apresentou como melhor marcador, seguido pela TGF-ß. Já o α-AML não se mostrou um bom sinalizador da evolução da reparação tissular.
Assuntos
Actinas/análise , Metaloproteinase 9 da Matriz/biossíntese , Ferida Cirúrgica , Fator de Crescimento Transformador beta/análise , Cicatrização , Actinas/fisiologia , Animais , Biomarcadores/análise , Imuno-Histoquímica , Masculino , Metaloproteinase 9 da Matriz/fisiologia , Ratos , Ratos Wistar , Ferida Cirúrgica/imunologia , Ferida Cirúrgica/patologia , Fator de Crescimento Transformador beta/fisiologiaRESUMO
RESUMO Objetivo: avaliar a eficácia de três marcadores imunoistoquímicos envolvidos no processo de cicatrização de ferida cirúrgica. Métodos: estudo experimental em 40 ratos da raça Wistar, dos marcadores metaloproteinases e metaloproteinase da matriz 9 (MMP-9), fator de transformação do crescimento beta (TGF-β) e miofibroblasto e alfa actina de músculo liso (α-AML), estudados a partir de fragmentos de cicatriz cirúrgica de incisão abdominal envolvendo pele, aponeurose e peritônio. Os animais foram distribuídos em quatro subgrupos de dez de acordo com o dia da morte, programada em três, sete, 14 e 21 dias. Resultados: na expressão da MMP-9 ocorreu aumento progressivo de sua concentração, mais evidente do 7º ao 14º dias variando a imuno-expressão tecidual entre 2,65% e 11,50%.TGF- β mostrou expressão em nível alto no 3º dia, caiu no 7º, voltando a subir no 14º, com pequena queda no 21º dia variando a imuno-expressão tecidual entre 0,03% e 2,92%. A α-AML apresentou níveis com pouca variação e discreto aumento variando a imuno-expressão tecidual entre 0,88% e 3,23%. Conclusão: a MMP-9 se apresentou como melhor marcador, seguido pela TGF-β. Já o α-AML não se mostrou um bom sinalizador da evolução da reparação tissular.
ABSTRACT Objective: to evaluate the efficacy of three immunohistochemical markers involved in the wound healing process. Methods: experimental study of 40 Wistar rats of the markers metalloproteinases and matrix metalloproteinase 9 (MMP-9), beta transforming growth factor (TGF-β) and myofibroblasts and smooth muscle actin alpha (α-MLA) markers, studied from fragments of surgical scar of abdominal incision involving skin, aponeurosis and peritoneum. The animals were divided into four subgroups of ten according to the day of death, scheduled in three, seven, 14 and 21 days. Results: MMP-9 expression showed a progressive increase of its concentration, more evident from 7th to 14th days, varying the tissue immunoexpression between 2.65% and 11.50% . TGF- β showed expression at high level on the 3rd day, fell in the 7th, rising again in the 14th, with a small decrease in the 21st day, varying the tissue immunoexpression between 0.03% and 2.92%. The α-AML presented levels with little variation and a slight increase, varying the tissue immunoexpression between 0.88% and 3.23%. Conclusion: MMP-9 presented as the best marker, followed by TGF-β. However, α-AML was not a good indicator of the evolution of tissue repair.