ABSTRACT
SUMMARY: Rheumatoid arthritis (RA) that affects the synovial knee joint causes swelling of the synovial membrane and tissue damage. Interleukin-17A (IL-17A) and the enzyme glycogen synthase kinase-3β (GSK3β) are involved in the pathogenesis of RA. The link between IL-17A, GSK3β, the oxidative stress, and the profibrogenic marker alpha-smooth muscle actin (α-SMA) with and without TDZD-8, GSK3β inhibitor has not been studied before. Consequently, active immunization of rats was performed to induce RA after three weeks using collagen type II (COII) injections. The treated group received daily injection of 1 mg/kg TDZD-8 for 21 days following the immunization protocol (COII+TDZD-8). Blood and synovium tissue samples were harvested at the end of the experiment. RA development was confirmed as corroborated by a substantial increase in blood levels of the highly specific autoantibody for RA, anti-citrullinated protein antibody as well as augmentation of reactive oxidative species (ROS) levels measured as lipid peroxidation. RA induction also increased synovium tissue levels of IL-17A and the profibrogenic marker, α-SMA. All these parameters seemed to be significantly (p<0.0001) ameliorated by TDZD-8. Additionally, a significant correlation between IL-17A, ROS, and α-SMA and biomarkers of RA was observed. Thus, knee joint synovium RA induction augmented IL-17A/GSK3β/ROS/α-SMA axis mediated arthritis in a rat model of RA, which was inhibited by TDZD-8.
La artritis reumatoide (AR) que afecta la articulación sinovial de la rodilla provoca inflamación de la membrana sinovial y daño tisular. La interleucina-17A (IL-17A) y la enzima glucógeno sintasa quinasa-3β (GSK3β) están involucradas en la patogenia de la AR. No se ha estudiadol vínculo entre IL-17A, GSK3β, el estrés oxidativo y el marcador profibrogénico actina de músculo liso alfa (α-SMA) con y sin inhibidor de TDZD-8, GSK3β. En consecuencia, se realizó una inmunización activa de ratas para inducir la AR después de tres semanas usando inyecciones de colágeno tipo II (COII). El grupo tratado recibió una inyección diaria de 1 µg/ kg de TDZD-8 durante 21 días siguiendo el protocolo de inmunización (COII+TDZD-8). Se recogieron muestras de sangre y tejido sinovial al final del experimento. El desarrollo de AR se confirmó como lo corroboró el aumento sustancial en los niveles sanguíneos del autoanticuerpo altamente específico para AR, el anticuerpo antiproteína citrulinada, así como el aumento de los niveles de especies oxidativas reactivas (ROS) medidos como peroxidación lipídica. La inducción de AR también aumentó los niveles de tejido sinovial de IL-17A y el marcador profibrogénico, α-SMA. Todos estos parámetros parecían mejorar significativamente (p<0,0001) con TDZD-8. Además, se observó una correlación significativa entre IL- 17A, ROS y α-SMA y biomarcadores de AR. Por lo tanto, la inducción de AR en la sinovial de la articulación de la rodilla aumentó la artritis mediada por el eje IL-17A/GSK3β/ROS/α-SMA en un modelo de rata de AR, que fue inhibida por TDZD-8.
Subject(s)
Animals , Rats , Arthritis, Rheumatoid , Thiadiazoles/administration & dosage , Fibrosis , Immunohistochemistry , Blotting, Western , Actins , Immunization , Reactive Oxygen Species , Rats, Wistar , Interleukin-17 , Collagen Type II/administration & dosage , Disease Models, Animal , Glycogen Synthase Kinase 3 betaABSTRACT
AIM OF WORK: To demonstrate the bleomycin induced histological changes in the lung and the possible protective and/or therapeutic effect of stem cell therapy. MATERIALS AND METHODS: Study was carried out on 36 adult male albino rats, classified into 4 groups: group I (control), group II (bleomycin treated group), group III (early stem cell treated group: immediately after bleomycin), group IV (late stem cell treated group: 7 days after bleomycin). Sections were taken at the 14th day of experiment. stained with Hematoxylin and Eosin, Masson's trichrome, immunohistochemichal stains for alpha-SMA & PCNA. Sections were examined by light & immunofluroscent microscopy. Area percent of collagen fibers, area percent & optical density of alpha-SMA immunopositive cells were measured as well as the number of H&E and PCNA stained pneumocytes type II was counted. RESULTS: Group II showed, thickening of septa, extravasation of blood, dividing pneumocytes type II cells with acinar formation, cellular infiltration, fibroblast cells, almost complete loss of normal lung architecture in certain fields, consolidation and replacement of the lung tissue with fibrous tissue in other fields. Restoring of lung tissue with significant decrease in mean area % of collagen fibers, alpha-SMA immunopositive cells were detected in group III. CONCLUSIONS: Early treatment with bone marrow derived mesenchymal stem cells (BMSCs) immediately after bleomycin administration showed a significant reduction in fibrotic changes, however the late treatment with BMSCs (7 days) after bleomycin administration showed non significant results.
Subject(s)
Adult , Animals , Humans , Male , Rats , Bleomycin , Bone Marrow , Collagen , Coloring Agents , Eosine Yellowish-(YS) , Fibroblasts , Hematoxylin , Lung , Mesenchymal Stem Cells , Microscopy , Alveolar Epithelial Cells , Proliferating Cell Nuclear Antigen , Pulmonary Fibrosis , Stem CellsABSTRACT
El cáncer cérvico uterino (CCU) es una patología de alta incidencia y mortalidad. La investigación hasta ahora se ha enfocado en estudiar su asociación con virus papiloma. Sin embargo, el estudio de la matriz extracelular (MEC) ha dado una nueva perspectiva para el estudio de factores inductores o perpetuadores de las neoplasias. En las neoplasias epiteliales como CCU el estroma tumoral presenta una composición dinámica de elementos celulares, destacando la presencia de miofibroblastos positivos a alfa actina de músculo liso (alfa SMA+) y fibrocitos CD34+. La MEC tiene un papel fundamental, ya que no sólo otorga las condiciones apropiadas para el desarrollo del tumor, sino que además condiciona el fenotipo de la población celular del estroma, donde la pérdida de fibrocitos CD34+ asociada a una ganancia de miofibroblastos alfa SMA+ podría ser un indicador muy sensible de invasión estromal, incluso en estadios iniciales. De la misma forma lo hace TGF-beta Ι, ya que su presencia es un reflejo de la síntesis de alfa SMA. Un nuevo elemento es versicán, un proteoglicano cuyas isoformas V0 y V1 se expresan también en tejidos neoplásicos de tumores ováricos, mama y cerebro, entre otros. Desempeña un papel muy importante en los fenómenos de adhesión celular, proliferación, migración y ensamblaje a la MEC. Por lo tanto, el análisis del estroma adyacente a las lesiones epiteliales del cuello uterino puede complementar el conocimiento sobre la conducta biológica de éstas, constituyendo una poderosa herramienta diagnóstica, de forma complementaria a los elementos utilizados hasta ahora.
Squamous cell carcinoma of the cervix (SCC) is a pathology that has high incidence and mortality. So far, research has been focused in the study of its association with papilloma virus. However, knowledge about extracellular matrix (ECM) has given a new perspective for the study of factors that induce or perpetuate neoplasms. In epithelial neoplasms like SCC, the tumoral stroma exhibits a dynamic composition of cellular elements, highlighting the presence of alpha actin of smooth muscle positive myofibroblasts (alpha SMA+) and CD34+ fibrocytes. ECM has an essential role, because it not only provides the appropriate conditions for tumor's development, but also affects stromal cell population phenotype, where a loss of CD34+ fibrocytes associated with a gain of alpha SMA+ myofibroblasts could be a sensitive indicator of stromal invasion, even in early stages. TGF-beta Ι does it in the same way, as its presence is a reflection of the synthesis of alpha SMA+. A new element is versican, a proteoglycan whose V0 and V1 isoforms expression is also observed in neoplastic tissues of ovary, breast and brain tumors, among others. It plays an important role in the phenomena of cellular adhesion, proliferation, migration and assembly of the ECM. Therefore, the analysis of the stroma adjacent to epithelial injuries of the cervix can complement the knowledge about the biological conducts of these, constituting a powerful diagnostic tool, as a complement to the elements used nowadays.
Subject(s)
Humans , Female , Extracellular Matrix , Uterine Cervical Neoplasms/pathology , Actins , Neoplasm Invasiveness , Stromal Cells , Transforming Growth Factor beta1 , VersicansABSTRACT
BACKGROUND/AIMS: Hepatic nerve innervation plays important roles in hepatic metabolism and hemodynamic mechanisms. We compared the distribution patterns of hepatic nerves between normal livers and two liver diseases to elucidate the effects of liver disease on the distribution of hepatic nerves. METHODS: Tissue specimens were obtained by ultrasonography-guided needle biopsies from 10 normal controls, 74 patients with chronic hepatitis (CH), and 35 patients with liver cirrhosis (LC). The obtained specimens were immunohistochemically stained using antibodies for S-100 protein and alpha-smooth-muscle actin (alpha-SMA). The degree of the expression in liver tissues was quantified by manual counting of positively stained nerve fibers under light microscopy. The serum hyaluronic acid level was assayed in all subjects to evaluate hepatic fibrosis. Electron microscopy examinations were also performed. RESULTS: The hepatic nerve innervation was significantly lower in LC than in normal controls, as indicated by S-100 protein staining. alpha-SMA and hyaluronic acid levels were higher in LC and CH than in normal controls. Electron microscopy revealed that unmyelinated nerve fiber bundles in the intralobar connective tissue coursed in the vicinity of hepatic triads. CONCLUSIONS: These results suggest that hepatic nerve innervation can be decreased by hepatic inflammatory responses and/or fibrotic changes in LC patients. Further study is needed to clarify this observation.
Subject(s)
Humans , Actins , Antibodies , Biopsy, Needle , Connective Tissue , Fibrosis , Hemodynamics , Hepatitis, Chronic , Hyaluronic Acid , Liver Cirrhosis , Liver Diseases , Liver , Metabolism , Microscopy , Microscopy, Electron , Nerve Fibers , Nerve Fibers, Unmyelinated , S100 ProteinsABSTRACT
BACKGROUND: There has been no general agreement in classifying basal cell carcinoma (BCC), and little is known about the immunohistochemical profiles in each subtypes of BCC. BCC is a locally-invasive tumor, but its aggressive forms tend to recur and metastasize. OBJECTIVE: In this study, we have compared the histolopathological subtypes of BCC by immunohistochemical study. We also focused on identifying representative markers of growth in the aggressive forms of BCC by assessing VEGF, p53 and alpha-SMA expression. METHODS: A total of 87 BCC specimens were collected at the 7 branch hospitals of The Catholic University of Korea from July 1997 to June 2003. For multiple immunohistochemical staining, a tissue microarray technique was used. The 87 samples were divided into 6 subtypes: 18 nodular, 19 nodular infiltrative, 12 micronodular, 14 infiltrative, 11 morphea and 13 basosquamous. Overall, 18 samples were classified as non-aggressive and the remaining 69 as aggressive. RESULTS: The following results were obtained after immunohistochemical staining with antibodies alpha-SMA, VEGF and p53. A significant increase of alpha-SMA expression was observed in aggressive forms of BCC, whereas the expression of p53, VEGF, the number of mast cells remained the same. The representative markers of tumor growth such as alpha-SMA were most highly expressed in the basosquamous type, and least expressed in the micronodular type compared to the nodular type. CONCLUSION: alpha-SMA was considered as an appropriate immunohistochemical marker in BCC to represent aggressiveness.
Subject(s)
Antibodies , Carcinoma, Basal Cell , Hospitals, Satellite , Immunohistochemistry , Korea , Mast Cells , Scleroderma, Localized , Vascular Endothelial Growth Factor AABSTRACT
BACKGROUND: Liver cirrhosis is a diffuse hepatic fibrosis and nodule formation. The transforming growth factor-beta1 (TGF-beta1) and interleukin-10 (IL-10) are very important cytokines in hepatic fibrogenesis. The aim of this study was to examine the relationship between the changes of the serum cytokines and morphological changes following common bile duct ligation in experimental rats. METHODS: Common bile ducts of fifty male Sprague-Dawley rats were ligated and seven male rats were set aside as controls. Five rats each were sacrificed in 1, 2, 4, 6, 8, and 10 experimental weeks. Light microscopic studies and liver function tests were performed during the above experimental weeks. The levels of serum TGF-beta1 and IL-10 were analyzed by ELISA. Also, alpha smooth muscle actin (alpha-SMA) immunohistochemical stains were performed. RESULTS: On the eighth week after common bile duct ligation, most hepatic lobular areas had been replaced by proliferated bile ducts and fibrous tissue (typical biliary cirrhosis). Serum TGF-beta1 levels between the control group and the common bile duct ligation group showed statistically significant changes. The alpha-SMA was stained at proliferated bile ducts. These findings were correlated with each other. CONCLUSION: Thus, this experiment may clarify our understanding of the mechanism in liver fibrogenesis. Also, indicated is a need to explore the therapeutic potential of these cytokines as anti-fibrotic agents.