RESUMO
BACKGROUND: Amid the COVID-19 pandemic, extensive testing was undertaken by independent clinical laboratories (ICLs), yet limited research exists on this matter. Drawing from Green Cross Laboratories (GC Labs)' pandemic response experience, this study seeks to offer insights for preparation for the next pandemic. METHODS: This retrospective study analyzed the outcomes of SARS-CoV-2 real-time reverse transcription polymerase chain reaction (SARS-CoV-2 rRT PCR) tests administered by GC Labs for COVID-19 diagnosis, upon request by different organizations, between February 2020 and April 2022. The distribution of institutions that requested the tests, the type of tests, and the positive rate were analyzed. We investigated resource allocation details. RESULTS: ICLs were responsible for conducting 85.6% of all tests carried out under South Korea's COVID-19 testing policy during the pandemic. The availability of free testing regardless of symptoms led to a significant increase in the use of pooled tests, which accounted for more than 80% of all tests conducted after August 2021. The gender and age distribution of COVID-19 cases nationwide and GC Labs' positive cases were similar. When we analyzed the positive rate by requesting organizations during the COVID-19 pandemic, despite an overall nationwide positivity rate of 35%, high-risk facilities exhibited a positivity rate of less than 5% by maintaining preemptive testing. The most notable increase in resources during the pandemic was seen in human resource input. CONCLUSIONS: South Korea's ICLs were able to conduct large volumes of testing during the COVID-19 pandemic because of their logistics and computer systems, scalable testing space, and trained testing personnel. They also had the flexibility to bring in additional resources to expand testing capacity because they are specialized testing organizations. Hence, ICLs could execute the pooled test that the government had introduced for extensive general population screening. The preemptive periodic testing of high-risk populations kept the positive rate much lower than in the general population. This study's findings will aid in refining mass testing-based policies for the next pandemic.
Assuntos
COVID-19 , Humanos , COVID-19/diagnóstico , COVID-19/epidemiologia , Teste para COVID-19 , SARS-CoV-2 , Laboratórios Clínicos , Pandemias/prevenção & controle , Estudos Retrospectivos , Técnicas de Laboratório Clínico , República da Coreia/epidemiologiaRESUMO
We investigated whether taurine chloramine (TauCl), which is -endogenously produced by immune cells such as macrophages that infiltrate adipose tissue, affects the differentiation of preadipocytes into adipocytes or modulates the expression of adipokines in adipocytes. To study the physiological effects of TauCl on human adipocyte differentiation and adipokine expression, preadipocytes were cultured under differentiation conditions for 14 days in the presence or the absence of TauCl. Differentiated adipocytes were also treated with TauCl in the presence or the absence of IL-1ß (1 ng/ml) for 7 days. The culture supernatants were analyzed for adipokines such as adiponectin, leptin, IL-6, and IL-8. At concentrations of 400-600 µM, TauCl significantly inhibited the differentiation of human preadipocytes into adipocytes in a dose-dependent manner. It did not induce the dedifferentiation of adipocytes or inhibit fat accumulation in adipocytes. Expression of major transcription factors of adipogenesis and adipocyte marker genes was decreased after treatment with TauCl, in agreement with its inhibition of -differentiation. These results suggest that TauCl may inhibit the differentiation of -preadipocytes into adipocytes. Thus, TauCl or more stable derivatives of TauCl could potentially be a safe drug therapy for obesity-related diseases.
Assuntos
Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Taurina/análogos & derivados , Adipócitos/metabolismo , Adipogenia/efeitos dos fármacos , Adipogenia/genética , Desdiferenciação Celular/efeitos dos fármacos , Desdiferenciação Celular/genética , Diferenciação Celular/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Especificidade de Órgãos/efeitos dos fármacos , Especificidade de Órgãos/genética , Taurina/farmacologiaRESUMO
Primary synovial cells with high passage number demonstrate increased production of proinflammatory mediators in response to inflammatory stimuli compared with cells with low passage number. This study used synovial cells to learn how different numbers of serial subculture passages affect the production of proinflammatory mediators in response to interleukin (IL)-1ß. Synovial cells were serially subcultured in flasks until passage 7. During cell passage, synovial cells were treated with IL-1ß for 24 h. Levels of proinflammatory mediators were analyzed by ELISA, real-time PCR, and Western blot. Synovial cells at passage 7 had elongated morphology and higher activity of ß-galactosidase, a marker of senescence, than those at passage 3. Production of IL-6, IL-8, vascular endothelial growth factor (VEGF), and prostaglandin E2 (PGE(2)) in response to IL-1ß was significantly increased in cells at passage 7 compared with passage 3. To evaluate the mechanism of this different response, IL-1 receptor expression was studied in cells stimulated with IL-1ß. Compared with cells at passage 3, cells at passage 7 had stronger expression of IL-1 receptors 1 and 2, as well as significantly increased expression of both receptors, in response to IL-1ß. This finding suggests that differential production in response to inflammatory stimuli associated with cell passages should be considered when in vitro experiments are used to evaluate the production levels of proinflammatory mediators.
Assuntos
Senescência Celular , Interleucina-1beta/farmacologia , Interleucina-8/biossíntese , Receptores de Interleucina-1/biossíntese , Membrana Sinovial/metabolismo , Fator A de Crescimento do Endotélio Vascular/biossíntese , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/metabolismo , Células Cultivadas , Dinoprostona/biossíntese , Humanos , Interleucina-6/biossíntese , Membrana Sinovial/citologia , Membrana Sinovial/efeitos dos fármacos , beta-Galactosidase/análiseRESUMO
We examined whether the expression and activation of pro-matrix metalloproteinase (MMP)-1 varies from that of pro-MMP-13 in the joint fluid of osteoarthritis (OA) and rheumatoid arthritis (RA) patients. To do this, joint fluid was collected from 34 RA and 34 OA patients. The collagenase (pro-MMP-1 and MMP-13, total MMP-1, and MMP-13), gelatinase (total MMP-2 and MMP-9), stromelysin (total MMP-3), matrilysin (total MMP-7), uPA, and tissue inhibitor of MMP (TIMP) levels were measured by ELISA. The level of total MMP-1 in RA joint fluids was similar to that of the OA joint fluid. In contrast, the level of total MMP-13 in the RA group was significantly higher than that of the OA group. Among various MMPs (MMP-2, MMP-3, MMP-7, and MMP-9), only MMP-9 was strongly associated with total MMP-13 in both RA and OA. The level of uPA was also strongly associated with MMP-13 in RA but not OA, while the level of TIMP-1 and TIMP-2 was not significantly different between RA and OA. In conclusion, MMP-9 and uPA might be involved in the activation of pro-MMP-13 through unknown mechanisms in arthritic diseases.
Assuntos
Artrite Reumatoide/enzimologia , Precursores Enzimáticos/análise , Metaloproteinase 13 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Osteoartrite/enzimologia , Líquido Sinovial/enzimologia , Ativador de Plasminogênio Tipo Uroquinase/análise , Adulto , Idoso , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Ativação Enzimática , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Metaloproteinase 1 da Matriz/análise , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 3 da Matriz/análise , Metaloproteinase 7 da Matriz/análise , Pessoa de Meia-Idade , Osteoartrite/tratamento farmacológico , Inibidores Teciduais de Metaloproteinases/análiseRESUMO
This study examined in an arthritis animal model whether elderly onset rheumatoid arthritis (EORA) is a more severe disease than younger onset rheumatoid arthritis. Arthritis was induced by injecting 5% kaolin/carrageenan into the left tibiotarsal ankles of 18-month-old and 4-week-old rats. Various parameters were measured to evaluate the arthritic progression of kaolin/carrageenan-induced arthritis in the rats. Immunohistochemical staining of arthritic joints was performed to determine the degree of inflammation in old and young rats. Measurements of ankle volume and thickness, arthritic index, number of squeaks, and the paw pressure test showed the 18-month-old rats had more severe disease than the young rats in a kaolin/carrageenan-induced arthritis model. The degree of inflammation and MMP-1 expression of arthritic joints in old rats was significantly higher than that of young rats based on histological evaluation with hematoxylin and eosin (H&E) staining and immunochemistry. More severe disease symptoms were found in old rats with EORA, but the molecular mechanisms still remain to be elucidated. Understanding the molecular mechanisms will be helpful to develop clinical protocols to efficiently treat patients with EORA, which is difficult to control with current protocols.
Assuntos
Artrite Experimental/patologia , Inflamação/patologia , Articulações/patologia , Acrilatos , Fatores Etários , Animais , Artrite Experimental/metabolismo , Carragenina , Inflamação/metabolismo , Articulações/metabolismo , Caulim , Metaloproteinase 1 da Matriz/metabolismo , Éteres Fenílicos , Ratos , Ratos Sprague-Dawley , Índice de Gravidade de DoençaRESUMO
In this study, data from a pandemic H1N1 outbreak in Korea were analyzed according to time, geography (districts), and age. A total of 252,271 samples collected nationwide were referred to the Greencross Reference Laboratory from June 2009 to February 2010 for H1N1 confirmation testing. Of these samples, 105,300 (41.7%) were H1N1-positive. With time, positivity was highest (57.0%) from October 26 - November 1 (4 weeks after Chuseok). The positive rates among districts show the highest value in Ulsan City (63.1%) and the lowest in Gyeongnam Province (32.8%). The positive rates for ages 0-2, 3-5, 6-11, 12-17, 18-20, 21-30, 31-40, 41-50, 51-60, and > 60 yr were 17.0%, 33.1%, 56.2%, 55.5%, 55.3%, 41.5%, 28.2%, 30.5%, 31.1%, and 16.8%, respectively, indirectly indicating propagation of H1N1 through schools. Pandemic control should involve school-targeted strategies.
Assuntos
Vírus da Influenza A Subtipo H1N1 , Influenza Humana/epidemiologia , Pandemias , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Influenza Humana/prevenção & controle , Masculino , Pessoa de Meia-Idade , Pandemias/prevenção & controle , Pandemias/estatística & dados numéricos , República da Coreia/epidemiologia , Estudantes , Adulto JovemRESUMO
This study was performed to provide evidence, albeit indirectly, as to which matrix metalloproteinases (MMPs), among the gelatinases MMP-2 and MMP-9 and the collagenases MMP-1 and MMP-13, play a more proactive role in the angiogenic process in arthritic joint. Joint fluid was collected from 33 patients with rhuematoid arthritis (RA) and osteoarthritis (OA), and protein (MMPs and vascular endothelial growth factor (VEGF)) levels were measured by ELISA, and the association of MMPs with VEGF was evaluated in joint fluid of patients with RA or OA. The levels of collagenases (total MMP-1 and total MMP-13) and gelatinases (total MMP-2 and total MMP-9) in RA joint fluid were significantly higher than those in OA fluid. Total MMP-9 levels were significantly associated with VEGF levels in RA fluids, but not in OA fluid, while total MMP-13 levels were strongly associated with VEGF levels in both RA and OA fluid. However, total MMP-2 and total MMP-1 levels were not associated with VEGF levels in either RA or OA joint fluid. Our results indirectly suggest that in RA and OA, MMP-9 and MMP-13 may play a more important role in angiogenesis than MMP-2 and MMP-1.
Assuntos
Artrite Reumatoide/metabolismo , Metaloproteinases da Matriz/análise , Osteoartrite/metabolismo , Líquido Sinovial/química , Humanos , Metaloproteinase 1 da Matriz/análise , Metaloproteinase 1 da Matriz/fisiologia , Metaloproteinase 13 da Matriz/análise , Metaloproteinase 13 da Matriz/fisiologia , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 2 da Matriz/fisiologia , Metaloproteinase 9 da Matriz/análise , Metaloproteinase 9 da Matriz/fisiologiaRESUMO
As a remedial option, the natural attenuation capacity of a petroleum contaminated groundwater at a military facility was examined. Hydrogeological conditions, such as high water level, permeable uppermost layer and frequent heavy rainfall, were favorable to natural attenuation at this site. The changes in the concentrations of electron acceptors and donors, as well as the relevant hydrochemical conditions, indicated the occurrence of aerobic respiration, denitrification, iron reduction, manganese reduction and sulfate reduction. The calculated BTEX expressed biodegradation capacity ranged between 20.52 and 33.67 mg/L, which appeared effective for the reduction of the contaminants levels. The contribution of each electron accepting process to the total biodegradation was in the order: denitrification > iron reduction > sulfate reduction > aerobic respiration > manganese reduction. The BTEX and benzene point attenuation rates were 0.0058-0.0064 and 0.0005-0.0032 day(-1), respectively, and the remediation time was 0.7-1.2 and 2.5-30 years, respectively. The BTEX and benzene bulk attenuation rates were 8.69 x 10(-4) and 1.05 x 10(-3) day(-1), respectively, and the remediation times for BTEX and benzene were 7.2 and 17.5 years, respectively. However, most of the natural attenuation occurring in this site can be attributed to dilution and dispersion. Consequently, the biodegradation and natural attenuation capacities were good enough to lower the contaminants levels, but their rates appeared to be insufficient to reach the remediation goal within a reasonable time frame. Therefore, some active remedial measures would be required.
Assuntos
Derivados de Benzeno/química , Água Subterrânea/química , Petróleo/análise , Poluentes Químicos da Água/química , Biodegradação Ambiental , Água Subterrânea/análise , Petróleo/metabolismo , República da CoreiaRESUMO
BACKGROUND: Adiponectin greatly stimulated the expression of matrix metalloproteinases (MMPs) in fibroblast-like synoviocytes (FLSs) as did IL-1beta. We wondered whether taurine chloramine (TauCl) inhibits the production of MMPs stimulated by adiponectin in the same pattern as by IL-1beta stimulation in vitro METHODS: Synovial cells from rheumatoid arthritis (RA) patients were treated with adiponectin or interleukin (IL)-1beta for 24 hr in the presence or absence of TauCl. The culture supernatant was collected and the levels of MMPs were measured by enzyme-linked immunosorbent assay (ELISA). The IkappaB signaling pathways stimulated by adiponectin were studied and the levels of NF-kappaB in the nuclei of the cells were analyzed by ELISA. RESULTS: TauCl (600 microM) inhibited MMP-13, but not MMP-1, expression in IL-1beta-stimulated RA FLSs. However, TauCl at the same concentration significantly inhibited the production of both adiponectin-stimulated MMP-1 and MMP-13 expression. TauCl inhibited the degradation of IkappaB-alpha stimulated by adiponectin, but not by IL-1beta. Similarly, the level of NF-kappaB in the nucleus was increased by adiponectin stimulation and was inhibited by 600 microM TauCl. However, the levels of NF-kappaB increased by IL-1beta stimulation were not inhibited by 600 microM TauCl. CONCLUSIONS: TauCl more effectively inhibited MMPs expression induced by adiponectin than that by IL-1beta in RA FLS, suggesting that TauCl plays an important role in down-regulating the expression of MMPs in arthritic joints.
Assuntos
Adiponectina/farmacologia , Inibidores Enzimáticos/farmacologia , Interleucina-1beta/farmacologia , Metaloproteinases da Matriz/biossíntese , Membrana Sinovial , Taurina/análogos & derivados , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Isoenzimas/biossíntese , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Membrana Sinovial/citologia , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/metabolismo , Taurina/farmacologiaRESUMO
Inflammation in the joint of rheumatoid arthritis is a complex immune reaction facilitated by various factors, such as cytokines, cells and hypoxia. Thus, we evaluated their relative capacity to produce proinflammatory mediators in response to IL-1beta, TNF-alpha or IL-17 under hypoxia or normoxia in fibroblast-like synoviocytes (FLSs) and macrophages. The level of IL-6 expression was strongly increased in both FLSs and THP-1 macrophages in response to IL-1beta and TNF-alpha, but the level by TNF-alpha was less than that by IL-1beta. In contrast, the expression of IL-8 in both cell types was strongly stimulated by both IL-1beta and TNF-alpha. In FLSs, PGE(2) production increased only in response to IL-1beta; and no effect was observed in THP-1 cells and TNF-alpha-stimulated FLSs. In addition, the production by IL-17 was extremely low when compared with those induced by IL-1beta or TNF-alpha in FLSs and THP-1 cells. Hypoxia (2% O(2)) decreased IL-1beta-stimulated production of PGE(2), even though it increased the expression of mRNA and protein of COX-2. These results suggest that IL-1beta and TNF-alpha differentially regulate gene expression in FLSs and macrophages under hypoxia or normoxia.
Assuntos
Dinoprostona/biossíntese , Fibroblastos/imunologia , Interleucina-1beta/fisiologia , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Macrófagos/imunologia , Membrana Sinovial/imunologia , Fator de Necrose Tumoral alfa/fisiologia , Linhagem Celular Tumoral , Células Cultivadas , Fibroblastos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Humanos , Macrófagos/metabolismo , Membrana Sinovial/citologia , Membrana Sinovial/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/imunologiaRESUMO
The gradual loss of telomeric DNA can contribute to replicative senescence and thus, having longer telomeric DNA is generally considered to provide a longer lifespan. Maintenance and stabilization of telomeric DNA is assisted by binding of multiple DNA-binding proteins, including those involved in double strand break (DSB) repair. We reasoned that declining DSB repair capacity and increased telomere shortening in aged individuals may be associated with decreased expression of DSB repair proteins capable of telomere binding. Our data presented here show that among the DSB repair proteins tested, only the expression of Ku70 and Mre11 showed statistically significant age-dependent changes in human lymphocytes. Furthermore, we found that expressions of Ku70 and Mre11 are statistically correlated, which indicate that the function of Ku70 and Mre11 may be related. All the other DSB repair proteins tested, Sir2, TRF1 and Ku80, did not show any significant differences upon aging. In line with these data, people who live in the regional community (longevity group), which was found to have statistically longer average life span than the rest area, shows higher level of Ku70 expression than those living in the neighboring control community. Taken together, our data show, for the first time, that Ku70 and Mre11 may represent new biomarkers for aging and further suggest that maintenance of higher expression of Ku70 and Mre11 may be responsible for keeping longer life span observed in the longevity group.
Assuntos
Envelhecimento/fisiologia , Antígenos Nucleares/metabolismo , Senescência Celular/fisiologia , Reparo do DNA/genética , Proteínas de Ligação a DNA/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Linfócitos T CD4-Positivos/metabolismo , DNA/genética , Humanos , Autoantígeno Ku , Longevidade , Proteína Homóloga a MRE11 , Pessoa de Meia-Idade , Telômero/genéticaRESUMO
The aim of this study was to determine whether disease activity and the type of therapy differentially modulate serum adipokine levels in patients with rheumatoid arthritis (RA), and whether pre-therapy adipokine levels contribute to resistance to treatment. Fasting blood samples from 40 RA patients were obtained at baseline and six months following therapeutic treatment with disease-modifying antirheumatic drugs (DMARDs) and/or tumor necrosis factor (TNF)-α blockers. Serum levels of adiponectin, leptin, visfatin and resistin were measured by ELISA. Baseline adipokine levels did not exhibit a statistically significant difference when comparing patients with moderate and high disease activity, based on the disease activity score in 28 joints (DAS28). Of all the adipokines, only adiponectin was significantly increased in patients responding to DMARDs and/or TNF-α blocker therapy, based on the American College of Rheumatology 20% improvement criteria (ACR20) at six months (2,964±1,237 to 3,683±1,511 ng/ml, P<0.01). However, adiponectin levels in non-responders did not significantly increase (3,192±2,090 to 3,222±1,150 ng/ml). By contrast, there were no statistically significant changes in leptin, resistin or visfatin levels in either the responders or non-responders. Serum adipokine (adiponectin, leptin, visfatin, and resistin) levels in RA patients did not significantly change following therapy, with the exception of adiponectin. Adipokine levels may not contribute to therapeutic resistance to DMARDs and/or TNF-α blocking agents.
Assuntos
Adipocinas/sangue , Artrite Reumatoide/sangue , Resistencia a Medicamentos Antineoplásicos , Adiponectina/sangue , Adulto , Idoso , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Leptina/sangue , Masculino , Pessoa de Meia-Idade , Nicotinamida Fosforribosiltransferase/sangue , Resistina/sangue , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismoRESUMO
WIN-34B showed analgesic and anti-inflammatory effects in various animal models of pain and osteoarthritis. However, the molecular mechanism by which WIN-34B inhibits pain and inflammation in vivo remains to be elucidated. We investigated the molecular mechanisms of the actions of WIN-34B using various in vitro models using fibroblast-like synoviocytes from patients with rheumatoid arthritis (RA FLSs), RAW264.7 cells and peritoneal macrophages. WIN-34B inhibited the level of IL-6, PGE2, and MMP-13 in IL-1ß-stimulated RA FLSs in a dose-dependent manner. The mRNA levels were also inhibited by WIN-34B. The level of PGE2, NO, IL-1ß, and TNF-α were inhibited by WIN-34B at different concentrations in LPS-stimulated RAW264.7 cells. The production of NO and PGE2 was inhibited by WIN-34B in a dose-dependent manner in LPS-stimulated peritoneal macrophages. All of these effects were comparable to the positive control, celecoxib or indomethacin. IκB signaling pathways were inhibited by WIN-34B, and the migration of NF-κB into the nucleus was inhibited, which is consistent with the degradation of IκB-α. Taken together, the results suggest that WIN-34B has potential as a therapeutic drug to reduce pain and inflammation by inhibiting the production of pro-inflammatory mediators.
RESUMO
To determine whether adiponectin may have synergistic effects in combination with the proinflammatory cytokine interleukin (IL)-1ß regarding the production of proinflammatory mediators during arthritic joint inflammation, synovial cells from rheumatoid arthritis (RA) patients were treated with adiponectin, IL-1ß, and their combination for 24 h. Culture supernatant was collected and analyzed by enzyme-linked immunosorbent assay for levels of IL-6, IL-8, prostaglandin E(2) (PGE(2)), vascular endothelial growth factor (VEGF), and matrix metalloproteinases (MMPs). Adiponectin-mediated intracellular signaling pathways were investigated to elucidate the molecular mechanisms underlying their synergy. The association of proinflammatory mediators with adiponectin was investigated in the synovial fluid of arthritis patients. Adiponectin functioned synergistically with IL-1ß to activate IL-6, IL-8, and PGE2 expression in RA fibroblast-like synoviocytes; Levels of VEGF, MMP-1, and MMP-13 were not synergistically stimulated. Adiponectin and IL-1ß each increased the expression of both adiponectin receptor 1 and IL-1 receptor 1. However, adiponectin and IL-1ß did not synergistically support the degradation of IκB-α or the nuclear translocation of NF-κB. Synergistically increased gene expression was significantly inhibited by MG132, an NF-κB inhibitor. Supporting the in vitro results, IL-6 and IL-8 levels were positively associated with adiponectin in synovial joint fluid from patients with RA, but not osteoarthritis (OA). In conclusion, adiponectin and IL-1ß may synergistically stimulate the production of proinflammatory mediators through unknown signaling pathways during arthritic joint inflammation. Adiponectin may be more important to the pathogenesis of RA than previously thought.
Assuntos
Adiponectina/metabolismo , Artrite Reumatoide , Inflamação , Interleucina-1beta/metabolismo , Líquido Sinovial , Adiponectina/administração & dosagem , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Células Cultivadas , Ciclo-Oxigenase 2/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/metabolismo , Inflamação/patologia , Interleucina-1beta/administração & dosagem , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Articulações/metabolismo , Articulações/patologia , Metaloproteinases da Matriz , NF-kappa B/metabolismo , Obesidade/metabolismo , Obesidade/patologia , Osteoartrite , Receptores de Adiponectina/metabolismo , Receptores de Interleucina-1/metabolismo , Líquido Sinovial/citologia , Líquido Sinovial/metabolismoRESUMO
The aim of this study was to evaluate whether thymosin ß4 (Tß4) levels are increased in the serum of rheumatoid arthritis (RA) patients, and if this increase is associated with RA disease activity and resistance to treatment. Blood samples from 40 patients with RA were collected at baseline and 6 months after starting treatment with disease-modifying antirheumatic drugs (DMARD) and/or tumor necrosis factor (TNF)-α blocker. Serum levels of Tß4 were measured by ELISA. Tß4 levels (mean ± standard deviation) in RA patients were significantly (approximately tenfold) higher than in healthy controls (577.4 ± 67.92 vs. 56.61 ± 5.72 ng/mL). Serum Tß4 levels in patients with severe disease activity before therapy were slightly higher than in patients with moderate disease activity (662.4 ± 491.5 vs. 462.5 ± 305.3 ng/ml, P > 0.05). Tß4 levels were significantly associated with disease activity according to the 28-joint Disease Activity Score. The mean Tß4 level at baseline in the DMARD treatment group was significantly lower than in the DMARD + TNF-α blocker treatment group. Tß4 levels were increased in the serum of patients with RA and were positively associated with disease activity. Levels of Tß4 may also be relevant in determining or predicting resistance to RA treatment. Further studies are necessary to determine if Tß4 is an appropriate therapeutic target for controlling inflammation associated with RA.
Assuntos
Antirreumáticos/uso terapêutico , Artrite Reumatoide/sangue , Timosina/sangue , Fator de Necrose Tumoral alfa/uso terapêutico , Adulto , Artrite Reumatoide/tratamento farmacológico , Resistência a Medicamentos , Quimioterapia Combinada , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Taheebo, the purple inner bark of the Bignoniaceae tree Tabebuia avellanedae Lorentz ex Griseb, which is found in tropical rain forests in northeastern Brazil, has been used as a traditional medicine for various diseases for more than 1,500 years. In the current study, various animal models were used to demonstrate the analgesic and anti-inflammatory properties of its ethanolic extract, thereby investigating its potential as a therapeutic treatment for diseases with pain and inflammation. In the hot plate and writhing tests for the in vivo analgesic effect test of Taheebo, a 200 mg/kg dose of the extract induced a significant anti-nociceptive effect and increased the pain threshold by approximately 30% compared with the control. In vascular permeability and tetradecanoylphorbol acetate (TPA), arachidonic acid- and carrageenan-induced paw edema tests for anti-inflammatory effects, treatment with 200 mg/kg Taheebo led to significant anti-inflammatory effects and inhibited inflammation by 30-50% compared with the control. At 100 mg/kg, the extract decreased the levels of pain and inflammation in all tested models, but the degree of inhibition was not statistically significant. The results suggest that the ethanolic extract of the inner bark of Tabebuia avellanedae has the potential to be developed as a therapeutic or supportive drug against diseases with accompanying pain and inflammation, including osteoarthritis.
Assuntos
Analgésicos/farmacologia , Anti-Inflamatórios/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Etanol/química , Limiar da Dor/efeitos dos fármacos , Extratos Vegetais/farmacologia , Tabebuia/química , Analgésicos/química , Analgésicos/uso terapêutico , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/uso terapêutico , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Masculino , Medicina Tradicional , Camundongos , Camundongos Endogâmicos ICR , Dor/tratamento farmacológico , Casca de Planta/química , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Ratos , Ratos Sprague-DawleyRESUMO
The aim of this study was to determine whether the inflammatory milieu and/or hypoxia induces the dedifferentiation of synovial cells into mesenchymal stem-like cells, which may contribute to the tumor-like growth of synovial cells. Expression of mesenchymal stem cell markers (CD24, CD44, CD90, CD106, CD146 and Stro-1) was compared among cultured fibroblast-like synoviocytes (FLSs) from patients with rheumatoid arthritis (RA) or osteoarthritis (OA), bone marrow mesenchymal stem cells (BM MSCs) and normal dermal fibroblasts. After the cells were stimulated with pro-inflammatory cytokines for 3 days under hypoxia or normoxia, the stem cell markers were analyzed by FACS. CD44 and CD90 were expressed constitutively in all four cell types. Only the BM MSCs strongly expressed CD146. The expression of stem cell markers was similar between FLSs from RA and those from OA patients. In addition, the expression levels in FLSs were similar to those in normal dermal fibroblasts. The stimulation of FLSs and dermal fibroblasts with IL-1ß or a mixture of cytokines under hypoxia did not induce a marked change in the expression of stem cell markers. These results indirectly suggest that the pro-inflammatory milieu may be not sufficient to induce the dedifferentiation of FLSs in arthritic joints.
Assuntos
Diferenciação Celular , Citocinas/farmacologia , Fibroblastos/citologia , Fibroblastos/metabolismo , Membrana Sinovial/citologia , Membrana Sinovial/metabolismo , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Células da Medula Óssea/citologia , Antígeno CD146/metabolismo , Hipóxia Celular , Células Cultivadas , Humanos , Receptores de Hialuronatos/metabolismo , Interleucina-1beta/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteoartrite/metabolismo , Osteoartrite/patologia , Membrana Sinovial/efeitos dos fármacos , Antígenos Thy-1/metabolismoRESUMO
Pyrrolidine dithiocarbamate (PDTC) can form a complex with metal ions and then act as a proteasome inhibitor, which leads to tumor cell apoptosis, and could therefore be developed as an anticancer agent. In our efforts to find factors that induce PDTC-mediated apoptosis of tumor cells, the effect of serum concentration on the apoptotic activity of PDTC was investigated. PDTC could not induce MCF-7 breast tumor cell death in serum-free media but significantly induced cell death in a dose-dependent manner at concentrations of ≥25 µM in media containing 10% fetal bovine serum. PDTC-mediated cell death was also dependent on serum concentration. PDTC-mediated cell death occurred through apoptosis. Similar to that in normal FBS, PDTC-mediated apoptotic cell death was also induced in media containing dialyzed FBS, indicating that PDTC-mediated apoptosis does not require metal ions or salts, but rather proteins in fetal bovine serum. In addition, differential apoptotic effects of PDTC were not observed with inhibitors of NF-κB activation such as N-acetylcysteine (NAC), Fenofibrate and carbobenzoxyl-l-leucyl-l-leucyl-l-leucinal (MG132) or with the metal-binding agent, 5-chloro-7-iodo-8-hydroxyquinoline (Clioquinol). These results indicate that serum is required for PDTC-mediated apoptosis and that zinc-binding compounds such as PDTC, N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) and Clioquinol may each have their own mechanisms by which they induce tumor cell death, even though they are all classified as zinc-binding compounds.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Sanguíneas/metabolismo , Neoplasias da Mama/tratamento farmacológico , Sangue Fetal/química , Pirrolidinas/farmacologia , Tiocarbamatos/farmacologia , Animais , Neoplasias da Mama/patologia , Bovinos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Quelantes/farmacologia , Meios de Cultura/química , Diálise , Inibidores Enzimáticos/farmacologia , Feminino , Humanos , NF-kappa B/antagonistas & inibidores , Inibidores de Proteassoma , Soro/química , Zinco/metabolismoRESUMO
Activated NF-kappaB plays an important role in the expression of matrix metalloproteinase (MMP)-1 and MMP-13 in rheumatoid arthritis and osteoarthritis. The objective of this study was to determine the effects of the NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC) on the expression of MMPs in IL-1beta-stimulated fibroblast-like synoviocytes (FLSs) of rheumatoid arthritis patients. FLSs were treated with IL-1beta (10 ng/ml) for 24 h in the presence or absence of PDTC. The level of MMP-1 and MMP-13 increased in response to PDTC in time- and dose-dependent manners in IL-1beta-stimulated FLSs; the expressions of IL-6 and vascular endothelial growth factor (VEGF) decreased in a PDTC concentration-dependent manner. However, PDTC-mediated repression of IL-6 and VEGF expression was not observed in TNF-alpha-stimulated rheumatoid arthritis FLSs. In contrast, other NF-kappaB inhibitors, such as fenofibrate, N-acetylcysteine and MG132, decreased MMP expression in IL-1beta-stimulated FLSs. The stimulatory effect of PDTC on MMP expression was not mimicked by specific inhibitors of the mitogen-activated protein kinase (MAPK) signaling pathway. Treatments with 100 muM PDTC did not inhibit the phosphorylation of p-ERK1/2, p-P38, and p-JNK, or the transnuclear migration of NF-kappaB through degradation of IkappaB-alpha in IL-1beta-stimulated FLSs. These results suggest that the increase of MMP expression may occur in a stimuli-specific manner or by an NF-kappaB independent mechanism. Therefore, therapeutic NF-kappaB inhibitors should be thoroughly studied before their clinical use in treating rheumatoid arthritis, as undesirable genes may be upregulated through unknown mechanisms, possibly resulting in worse symptoms.
Assuntos
Artrite Reumatoide/patologia , Interleucina-1beta/farmacologia , Metaloproteinases da Matriz/biossíntese , NF-kappa B/antagonistas & inibidores , Pirrolidinas/farmacologia , Membrana Sinovial/patologia , Tiocarbamatos/farmacologia , Regulação para Cima/efeitos dos fármacos , Animais , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Fibroblastos/patologia , Humanos , Proteínas I-kappa B/metabolismo , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/metabolismo , Osteoartrite/patologia , Coelhos , Transdução de Sinais/efeitos dos fármacos , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/metabolismo , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Mizoribine is a disease-modifying anti-rheumatic drug (DMARD) that is used in the treatment of rheumatoid arthritis. However, clinical use of the drug is restricted to a few Asian countries due to a lack of comprehensive evidence on its effectiveness. The inhibitory effect of the drug on human osteoclastogenesis was investigated in the hopes of providing some clear evidence. Mizoribine was found to inhibit in vitro osteoclastogenesis in a dose-dependent manner. In addition, the size of the pit area was closely related to the number of osteoclasts in a bone resorption assay. However, mizoribine did not affect the phosphorylation of MAP kinase (p38, JNK, ERK), the degradation of IkappaBalpha, or receptor activator of NF-kappaB ligand (RANKL) expression in fibroblast-like synoviocytes stimulated with IL-1beta. These results suggested that mizoribine may partially suppress osteoclastogenesis, leading to progressive bone erosion by inhibiting the growth or the signaling pathway of precursor cells to form osteoclasts rather than fibroblast-like synoviocytes.