RESUMO
A known consequence of the large weight loss after bariatric surgery is the appearance of large skinfolds, particularly in the abdomen region of the patients. The balance between the synthesis of extracellular matrix (ECM) components and their proteolysis, mainly by fibrinolytic systems and matrix metalloproteases (MMPs), may be disturbed in these patients. The causes underlying the deregulation of ECM remodeling that occurs in these patients are not, however, clear. We investigated molecular mechanisms responsible for this dysfunction of ECM remodeling process, comparing it to normal skin. Collagen types, MMP2 and MMP9 expression and activity, interleukins 1ß (IL1ß) and 6 (IL6), and transcription coactivator PGC-1ß expression were analyzed in 16 patients. Ex-obese patients presented increased expression of collagen types III and IV mRNA, increased expression of MMP2, decreased expression and activity of MMP9, and increased expression of PGC-1ß in the skin. Inflammation markers IL1ß and IL6 mRNA were not different. We have demonstrated that obese patients with extensive weight loss after bariatric surgery have increased expression of PGC-1ß in the skin, which can result in a decreased expression and activity of MMP9 and increased collagen types III and IV deposition. These molecular changes may contribute for the formation of saggy skinfolds observed in these patients and impair wound healing.
Assuntos
Matriz Extracelular/metabolismo , Obesidade/metabolismo , Pele/metabolismo , Redução de Peso , Cirurgia Bariátrica , Colágeno Tipo III/biossíntese , Colágeno Tipo IV/biossíntese , Matriz Extracelular/patologia , Feminino , Regulação da Expressão Gênica , Humanos , Interleucina-1beta/biossíntese , Interleucina-6/biossíntese , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Pessoa de Meia-Idade , Obesidade/patologia , Obesidade/cirurgia , Pele/patologiaRESUMO
The aim of this study was to determine the effect of artesunate on extracellular matrix (ECM) accumulation and the expression of collagen-IV, matrix metalloproteinase (MMP), and tissue inhibitor of matrix metalloproteinase (TIMP) to understand the pharmacological role of artesunate in pulmonary fibrosis. Eighty Sprague-Dawley rats were randomly assigned to four groups that were administered saline alone, bleomycin (BLM) alone, BLM + artesunate, or artesunate alone for 28 days. Lung tissues from 10 rats in each group were used to obtain lung fibroblast (LF) primary cells, and the rest were used to analyze protein expression. The mRNA expression of collagen-IV, MMP-2, MMP-9, TIMP-1, and TIMP-2 in lung fibroblasts was detected by real-time quantitative reverse transcriptase polymerase chain reaction. The protein levels of collagen-IV, MMP-2, MMP-9, TIMP-1, and TIMP-2 protein in lung tissues were analyzed by western blotting. Artesunate treatment alleviated alveolitis and pulmonary fibrosis induced by bleomycin in rats, as indicated by a decreased lung coefficient and improvement of lung tissue morphology. Artesunate treatment also led to decreased collagen-IV protein levels, which might be a result of its downregulated expression and increased MMP-2 and MMP-9 protein and mRNA levels. Increased TIMP-1 and TIMP- 2 protein and mRNA levels were detected after artesunate treatment in lung tissues and primary lung fibroblast cells and may contribute to enhanced activity of MMP-2 and -9. These findings suggested that artesunate attenuates alveolitis and pulmonary fibrosis by regulating expression of collagen-IV, TIMP-1 and 2, as well as MMP-2 and -9, to reduce ECM accumulation.
Assuntos
Colágeno Tipo IV/biossíntese , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Fibrose Pulmonar/tratamento farmacológico , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-2/biossíntese , Animais , Artemisininas/administração & dosagem , Artesunato , Colágeno Tipo IV/genética , Modelos Animais de Doenças , Matriz Extracelular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Metaloproteinase 2 da Matriz/genética , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , RNA Mensageiro/biossíntese , Ratos , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-2/genéticaRESUMO
PURPOSE: Keratoconus (KTCN) is a non-inflammatory, usually bilateral disorder of the eye which results in the conical shape and the progressive thinning of the cornea. Several studies have suggested that genetic factors play a role in the etiology of the disease. Several loci were previously described as possible candidate regions for familial KTCN; however, no causative mutations in any genes have been identified for any of these loci. The purpose of this study was to evaluate role of the collagen genes collagen type IV, alpha-1 (COL4A1) and collagen type IV, alpha-2 (COL4A2) in KTCN in Ecuadorian families. METHODS: COL4A1 and COL4A2 in 15 Ecuadorian KTCN families were examined with polymerase chain reaction amplification, and direct sequencing of all exons, promoter and intron-exon junctions was performed. RESULTS: Screening of COL4A1 and COL4A2 revealed numerous alterations in coding and non-coding regions of both genes. We detected three missense substitutions in COL4A1: c.19G>C (Val7Leu), c.1663A>C (Thr555Pro), and c.4002A>C (Gln1334His). Five non-synonymous variants were identified in COL4A2: c.574G>T (Val192Phe), c.1550G>A (Arg517Lys), c.2048G>C (Gly683Ala), c.2102A>G (Lys701Arg), and c.2152C>T (Pro718Ser). None of the identified sequence variants completely segregated with the affected phenotype. The Gln1334His variant was possibly damaging to protein function and structure. CONCLUSIONS: This is the first mutation screening of COL4A1 and COL4A2 genes in families with KTCN and linkage to a locus close to these genes. Analysis of COL4A1 and COL4A2 revealed no mutations indicating that other genes are involved in KTCN causation in Ecuadorian families.
Assuntos
Colágeno Tipo IV/genética , Ceratocone/genética , Sequência de Aminoácidos , Estudos de Casos e Controles , Colágeno Tipo IV/análise , Colágeno Tipo IV/biossíntese , Córnea/patologia , Equador , Feminino , Perfilação da Expressão Gênica , Haplótipos , Humanos , Masculino , Dados de Sequência Molecular , Mutação , Linhagem , Fenótipo , Polimorfismo de Nucleotídeo Único , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Obesity is a risk factor for the development of chronic kidney disease (CKD) and end-stage renal disease. It is not clear whether the adoption of a high-protein diet in obese patients affects renal lipid metabolism or kidney function. Thus the aims of this study were to assess in obese Zuckerfa/fa rats the effects of different types and amounts of dietary protein on the expression of lipogenic and inflammatory genes, as well as renal lipid concentration and biochemical parameters of kidney function. Rats were fed different concentrations of soy protein or casein (20, 30, 45%) for 2 mo. Independent of the type of protein ingested, higher dietary protein intake led to higher serum triglycerides (TG) than rats fed adequate concentrations of protein. Additionally, the soy protein diet significantly increased serum TG compared with the casein diet. However, rats fed soy protein had significantly decreased serum cholesterol concentrations compared with those fed a casein diet. No significant differences in renal TG and cholesterol concentrations were observed between rats fed with either protein diets. Renal expression of sterol-regulatory element binding protein 2 (SREBP-2) and its target gene HMG-CoA reductase was significantly increased as the concentration of dietary protein increased. The highest protein diets were associated with greater expression of proinflammatory cytokines in the kidney, independent of the type of dietary protein. These results indicate that high soy or casein protein diets upregulate the expression of lipogenic and proinflammatory genes in the kidney.
Assuntos
Caseínas/administração & dosagem , Proteínas Alimentares/administração & dosagem , Rim/fisiologia , Obesidade/metabolismo , Proteínas de Soja/administração & dosagem , Animais , Glicemia/metabolismo , Caseínas/farmacologia , Colesterol/sangue , Colágeno Tipo IV/biossíntese , Proteínas Alimentares/farmacologia , Peróxido de Hidrogênio/urina , Hidroximetilglutaril-CoA Redutases/biossíntese , Insulina/sangue , Interleucina-6/biossíntese , Rim/anatomia & histologia , Rim/efeitos dos fármacos , Lipogênese , Tamanho do Órgão , Estresse Oxidativo , Ratos , Ratos Zucker , Proteínas de Soja/farmacologia , Proteína de Ligação a Elemento Regulador de Esterol 1/biossíntese , Proteína de Ligação a Elemento Regulador de Esterol 2/biossíntese , Fator de Crescimento Transformador beta/biossíntese , Triglicerídeos/sangue , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Ameloblastomatous epithelium containing clusters of ghost cells is the typical histopathology of calcifying cystic odontogenic tumor (CCOT). This paper aimed to assess keratins AE1-AE3, K7, K10/13, K14, K18, K19, vimentin, laminin, and collagen IV in 08 CCOTs to discuss their histopathogenesis. Similarity to the immunoprofile of the stratified squamous epithelium was seen in the with the basal layer expressing K14 and the upper cells expressing K10/13. When compared to the immunoprofile of the normal odontogenic epithelium, of odontogenic tumor epithelia and of the ghost cells described in the literature, it was possible to suggest that the CCOT epithelium differentiates towards squamous type.
Assuntos
Neoplasias Maxilomandibulares/metabolismo , Neoplasias Maxilomandibulares/patologia , Queratinas/biossíntese , Cisto Odontogênico Calcificante/metabolismo , Cisto Odontogênico Calcificante/patologia , Diferenciação Celular , Transformação Celular Neoplásica , Colágeno Tipo IV/biossíntese , Epitélio/metabolismo , Humanos , Imuno-Histoquímica , Laminina/biossíntese , Vimentina/biossínteseRESUMO
The objective of this paper was to study the morphology of the articular disc and analyze the immunohistochemical expression of the marker of type IV collagen in the articular disc of the temporomandibular joint (TMJ) of human fetuses of different gestational ages. Twenty TMJ from human fetuses aging from 21 to 24 weeks of intrauterine life were studied. The TMJ were supplied by the Federal University of Uberaba. The ages of the fetuses were determined by measuring the crown-rump length (CRL). Macroscopically, the fetuses were fixed in a formalin solution at 10% and dissected by removing the skin and the subcutaneous tissue, exposing the deep structures. An immunohistochemical marker of type IV collagen was used in order to characterize the presence of blood vessels in the central region of the temporomandibular joint disc. Analysis of the immunohistochemical marker of type IV collagen showed the presence of blood vessels in the central region of the temporomandibular disc in human fetuses.
Assuntos
Vasos Sanguíneos/embriologia , Colágeno Tipo IV/biossíntese , Neovascularização Fisiológica/fisiologia , Disco da Articulação Temporomandibular/irrigação sanguínea , Disco da Articulação Temporomandibular/embriologia , Especificidade de Anticorpos , Biomarcadores/análise , Biomarcadores/metabolismo , Vasos Sanguíneos/citologia , Vasos Sanguíneos/metabolismo , Colágeno Tipo IV/análise , Feto , Idade Gestacional , Humanos , Imuno-Histoquímica , Disco da Articulação Temporomandibular/metabolismoRESUMO
This study evaluates the effect of poly(L-lactic acid) (PLLA) and poly(hydroxybutyrate-cohydroxyvalerate) (PHBV) bioabsorbable polymers and their blends on the induction of alteration of cell growth pattern in vitro. Vero cells were cultured on PLLA, PHBV, and different blends (100/0, 60/40, 50/50, 40/60, and 0/100). The cell adhesion assay showed that the best results were obtained with the (60/40, 50/50) blends. Scanning electron microscopy showed that the cells on (100/0) and (60/40) samples grew with a round morphology preferentially in the porous areas. The (50/50) blends had cells in the porous and smooth areas in a similar way. The (40/60) blends showed spreading cells on the smooth areas. The (0/100) sample, which had no pores, had spreading cells interconnected by filaments. Histological sections showed a confluent cell monolayer and the immunocytochemistry showed that the cells produced collagen IV and fibronectin on all substrates. Thus, we conclude that PLLA/PHBV blends were efficient in maintaining cell growth and producing an extracellular matrix on them.