Perfil de IL-6 e TNF na Formação de Células Espumosas: Um Método Aprimorado Usando a Sonda de Isotiocianato de Fluoresceína (FITC) / Profile of IL-6 and TNF in Foam Cell Formation: An Improved Method Using Fluorescein Isothiocyanate (FITC) Probe

Resumo Fundamento A formação de células espumosas ocorre devido ao aumento em lipoproteína plasmática de baixa densidade (LDL) e desregulação da inflamação, sendo importante para o desenvolvimento da aterosclerose. Objetivo Avaliar o perfil do fator de necrose tumoral alfa (TNF-α) e da interleucina-6 (IL-6) no método de formação da célula espumosa existente, otimizando esse protocolo. Métodos A LDL foi isolada, oxidada e marcada com sonda de isotiocianato de fluoresceína (FITC). As células espumosas foram geradas de célula derivada de monócitos humanos THP-1 e incubadas na ausência (controle) ou presença de FITC-ox-LDL (10, 50, 100, 150 ou 200 μg/mL), por 12, 24, 48 ou 72 horas. A FITC-ox-LDL na célula foi quantificada por microscopia. O ensaio de imunoabsorção enzimática foi avaliado para quantificar a IL-6 e o TNF-α, com um p <0,05 considerado significativo. Resultados Todas as concentrações de FITC-ox-LDL testadas apresentaram fluorescência mais alta em comparação com o controle, demonstrando maior acúmulo de lipoproteínas nas células. Quanto mais alta a concentração de FITC-ox-LDL, maior a produção de TNF-α e IL-6. A produção de IL-6 pelas células espumosas foi detectada até o valor de 150 µg/mL da LDL máxima de estímulo. Concentrações acima de 50 μg/mL de LDL estimularam maior liberação de TNF-α comparado ao controle. Conclusões Nosso modelo contribui para o entendimento da liberação de IL-6 e TNF-α em resposta a várias concentrações de ox-LDL usando o método otimizado para a formação de células espumosas.

Abstract Background The formation of foam cells occurs due to the increase in low-density plasma lipoprotein (LDL) and dysregulation of inflammation, which is important for the development of atherosclerosis. Objective To evaluate the profile of tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) in the existing foam cell formation method, optimizing this protocol. Methods The LDL was isolated, oxidized, and labeled with a Fluorescein isothiocyanate (FITC) probe. Foam cells were generated from THP-1 human monocyte-derived cells and incubated in the absence (control) or presence of FITC-ox-LDL (10, 50, 100, 150, or 200 μg/mL), for 12, 24, 48, or 72 hours. The accumulated FITC-ox-LDL in the cell was quantified by microscopy. The enzyme-linked immunosorbent assay was evaluated to quantify the IL-6 and TNF-α, with p < 0.05 considered significant. Results All the FITC-ox-LDL concentrations tested showed a higher fluorescence when compared to the control, showing a greater accumulation of lipoprotein in cells. The higher the concentration of FITC-ox-LDL, the greater the production of TNF-α and IL-6. The production of IL-6 by foam cells was detected up to the value of 150 µg/mL of the maximum stimulus for LDL. Concentrations above 50 μg/mL LDL stimulated greater release of TNF-α compared to control. Conclusions Our model contributes to the understanding of the release of IL-6 and TNF-α in response to different concentrations of ox-LDL, using an optimized method for the formation of foam cells.

Profile of IL-6 and TNF in Foam Cell Formation: An Improved Method Using Fluorescein Isothiocyanate (FITC) Probe. / Perfil de IL-6 e TNF na Formação de Células Espumosas: Um Método Aprimorado Usando a Sonda de Isotiocianato de Fluoresceína (FITC).

BACKGROUND: The formation of foam cells occurs due to the increase in low-density plasma lipoprotein (LDL) and dysregulation of inflammation, which is important for the development of atherosclerosis. OBJECTIVE: To evaluate the profile of tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) in the existing foam cell formation method, optimizing this protocol. METHODS: The LDL was isolated, oxidized, and labeled with a Fluorescein isothiocyanate (FITC) probe. Foam cells were generated from THP-1 human monocyte-derived cells and incubated in the absence (control) or presence of FITC-ox-LDL (10, 50, 100, 150, or 200 µg/mL), for 12, 24, 48, or 72 hours. The accumulated FITC-ox-LDL in the cell was quantified by microscopy. The enzyme-linked immunosorbent assay was evaluated to quantify the IL-6 and TNF-α, with p < 0.05 considered significant. RESULTS: All the FITC-ox-LDL concentrations tested showed a higher fluorescence when compared to the control, showing a greater accumulation of lipoprotein in cells. The higher the concentration of FITC-ox-LDL, the greater the production of TNF-α and IL-6. The production of IL-6 by foam cells was detected up to the value of 150 µg/mL of the maximum stimulus for LDL. Concentrations above 50 µg/mL LDL stimulated greater release of TNF-α compared to control. CONCLUSIONS: Our model contributes to the understanding of the release of IL-6 and TNF-α in response to different concentrations of ox-LDL, using an optimized method for the formation of foam cells.

FUNDAMENTO: A formação de células espumosas ocorre devido ao aumento em lipoproteína plasmática de baixa densidade (LDL) e desregulação da inflamação, sendo importante para o desenvolvimento da aterosclerose. OBJETIVO: Avaliar o perfil do fator de necrose tumoral alfa (TNF-α) e da interleucina-6 (IL-6) no método de formação da célula espumosa existente, otimizando esse protocolo. MÉTODOS: A LDL foi isolada, oxidada e marcada com sonda de isotiocianato de fluoresceína (FITC). As células espumosas foram geradas de célula derivada de monócitos humanos THP-1 e incubadas na ausência (controle) ou presença de FITC-ox-LDL (10, 50, 100, 150 ou 200 µg/mL), por 12, 24, 48 ou 72 horas. A FITC-ox-LDL na célula foi quantificada por microscopia. O ensaio de imunoabsorção enzimática foi avaliado para quantificar a IL-6 e o TNF-α, com um p <0,05 considerado significativo. RESULTADOS: Todas as concentrações de FITC-ox-LDL testadas apresentaram fluorescência mais alta em comparação com o controle, demonstrando maior acúmulo de lipoproteínas nas células. Quanto mais alta a concentração de FITC-ox-LDL, maior a produção de TNF-α e IL-6. A produção de IL-6 pelas células espumosas foi detectada até o valor de 150 µg/mL da LDL máxima de estímulo. Concentrações acima de 50 µg/mL de LDL estimularam maior liberação de TNF-α comparado ao controle. CONCLUSÕES: Nosso modelo contribui para o entendimento da liberação de IL-6 e TNF-α em resposta a várias concentrações de ox-LDL usando o método otimizado para a formação de células espumosas.

Nandrolone increases angiotensin-I converting enzyme activity in rats tendons / Nandrolona aumenta a atividade da enzima conversora de angiotensina em tendões de ratos / La nandrolona aumenta la actividad de la enzima de conversión angiotensina en tendones de ratones

INTRODUCTION: The renin-angiotensin system (RAS) has been associated with several biological processes of the human body, regulating, among others blood pressure and water and electrolytes balance. Moreover, RAS also regulates connective tissue growth. Recently, studies have shown that the use of nandrolone modifies the angiotensin-I converting enzyme (ACE) activity and increases collagen deposition in the heart. OBJECTIVE: The aim of study was to evaluate the Angiotensin-I converting enzyme (ACE) activity in the superficial flexor tendon (SFT) and in serum after load exercise in combination with anabolic androgenic steroid (AAS) administration after training session and six weeks of detraining. METHODS: Forty-eight Wistar rats were used into two groups (G1 and G2) subdivided into four subgroups: Sedentary (S); trained (T); AAS-treated (Deca-Durabolin(r), 5mg/kg, twice a week) sedentary rats (AAS) and AAS-treated and trained animals (AAST). Trained groups performed jumps in water: four series of 10 jumps each, followed by a 30 sec interval between the series, for seven weeks. RESULTS: Training increased ACE activity in the SFT compared to the control group (p <0.05). Both AAS and AAST groups presented higher ACE activity levels (p < 0.05). The AAST increased the ACE activity only compared to the trained animals. Only the AAST group presented significant higher levels of ACE in the serum. In the G2 group, all experimental groups presented decreased ACE activity in the serum and in the tendon, as compared to the control group. CONCLUSION: This study indicates that AAS administration and its combination with exercise increased ACE activity of tendons. AAS abuse could compromise tendon adaptation causing maladaptive remodeling. .

INTRODUÇÃO: O sistema renina-angiotensina (SRA) tem sido associado a importantes processos biológicos do corpo humano, regulando, entre outros processos, a pressão arterial e balanço hidroeletrolítico. Além disso, o SRA também regula o crescimento do tecido conjuntivo. Recentemente, foi demonstrado que a utilização de nandrolona modifica a atividade da enzima conversora de angiotensina (ECA) e aumenta a deposição de colágeno no coração. OBJETIVO: O objetivo do estudo foi avaliar a atividade de ECA no tendão flexor superficial (TFS) e no soro após exercício de força com administração de esteroides anabólicos androgênicos (EAA) durante sete semanas e após seis semanas de destreinamento. MÉTODOS: Quarenta e oito ratos da linhagem Wistar foram divididos em dois grupos (G1 e G2) e subdivididos em quatro subgrupos: Sedentários (S); treinados (T); sedentários com EAA (EAAS) (Deca-Durabolin(r) - 5mg/kg, duas vezes por semana) e treinados com administração de EAA (EAAT). Os grupos treinados realizaram saltos na água: quatro séries de 10 saltos cada, com intervalo de 30 seg entre as séries. RESULTADOS: O treinamento aumentou a atividade de ECA no TFS em comparação ao controle (p<0,05). Os grupos tratados com EAA apresentaram maiores níveis de ECA (p<0,05). O grupo EAA-T mostrou atividade de ECA mais elevada quando comparada ao grupo T. Além disso, o grupo EAA-T apresentou maiores níveis de ECA no soro. No grupo G2, todos os subgrupos diminuíram a atividade de ECA tanto no soro quanto no tendão. CONCLUSÃO: Este estudo indica que a administração de EAA e sua combinação com o exercício aumenta a atividade de ECA nos tendões. O uso abusivo de EAA pode comprometer a adaptação tendínea no qual pode provocar remodelamento mal adaptativas. .

INTRODUCCIÓN: El sistema renina-angiotensina (RAS) ha sido asociado con varios procesos biológicos del cuerpo humano, entre ellos, regular la presión arterial y el contenido de electrolitos. Además, el RAS también regula el tejido conectivo. Recientemente, estudios han demostrado que el uso de nandrolona modifica la actividad de ACE e incrementa la deposición de colágeno en el corazón. OBJETIVO: En este modo, el objetivo del estudio fue evaluar la actividad de la enzima de conversión angiotensina (ACE) en el tendón flexor superficial (TFS) y en el suero después del ejercicio de resistencia en combinación con la administración de esteroides anabólico-androgénicos (AAS) después de la sesión de entrenamiento, y seis semanas de desentrenamiento. MÉTODOS: Cuarenta y ocho ratones Wistar fueron divididos en dos grupos (G1 y G2) y subdivididos en cuatro grupos: sedentarios (S); entrenados (T); ratas sedentarias tratadas con AAS (Deca-Durabolin(r) - 5 mg / kg dos veces a la semana) (AAS) y animales entrenados y tratados con AAS (AAST). Los grupos entrenados realizaron saltos en el agua: cuatro series de 10 saltos cada uno, con 30 segundos de intervalo entre las series, durante siete semanas. RESULTADOS: El entrenamiento aumentó la actividad de ECA en TFS en comparación con el control (p <0,05). Los grupos AAS y AAST mostraron mayores niveles de ACE (p <0,05). El grupo AAST mostró alta actividad de ECA en comparación con el grupo T. Además, el AAST mostró niveles más altos de ACE en el suero. En G2, todos los grupos disminuyeron la la actividad ACE tanto en el suero como en el tendón si comparados con el grupo control. CONCLUSIÓN: Este estudio indica que la administración de AAS y su combinación con el ejercicio aumenta la actividad de ECA en los tendones. El uso abusivo de AAS puede comprometer la adaptación del tendón, lo que puede causar remodelaciones mal adaptativas. .

Resistance training improves body composition and increases matrix metalloproteinase 2 activity in biceps and gastrocnemius muscles of diet-induced obese rats.

OBJECTIVE: We investigated the influence of resistance training on body composition and matrix metalloproteinase 2 activity in skeletal muscles of rats fed a high-fat diet. METHODS: Thirty-two Wistar rats were divided into four experimental groups (n = 8/each) according to diet and exercise status: Control (standard diet), Obese Control (high-fat diet), Resistance Training (standard diet) and Obese Resistance Training (high-fat diet) groups. Animals were fed a high-fat diet for 12 weeks to promote excessive weight gain. Resistance Training groups performed 12 weeks of training periods after this period in a vertical ladder three times/week. Fat percentage, fat-free mass and fat mass were assessed using dual-energy X-ray absorptiometry, and matrix metalloproteinase 2 activity in biceps and gastrocnemius muscles was analyzed using zymography. RESULTS: Resistance training significantly reduced body and fat masses and fat percentages in both trained groups (p<0.05). The maximal carrying load between trained groups was not different, but relative force was higher in the Resistance Training group (p<0.05). Of note, increased matrix metalloproteinase 2 activity was noted in the tested muscles of both trained groups (p<0.05). CONCLUSION: In conclusion, altered body composition and muscle matrix metalloproteinase 2 activity promoted by excessive weight gain were positively modified by resistance training.

The effects of exercise modalities on adiposity in obese rats

OBJECTIVE: The aim of the present study was to evaluate the effect of both swimming and resistance training on tumor necrosis factor-alpha and interleukin-10 expression, adipocyte area and lipid profiles in rats fed a high-fat diet. METHODS: The study was conducted over an eight-week period on Wistar adult rats, who were divided into six groups as follows (n = 10 per group): sedentary chow diet, sedentary high-fat diet, swimming plus chow diet, swimming plus high-fat diet, resistance training plus chow diet, and resistance training plus high-fat diet. Rats in the resistance training groups climbed a vertical ladder with weights on their tails once every three days. The swimming groups swam for 60 minutes/day, five days/week. RESULTS: The high-fat diet groups had higher body weights, a greater amount of adipose tissue, and higher tumor necrosis factor-alpha expression in the visceral adipose tissue. Furthermore, the high-fat diet promoted a negative change in the lipid profile. In the resistance training high-fat group, the tumor necrosis factor-alpha expression was lower than that in the swimming high-fat and sedentary high-fat groups. Moreover, smaller visceral and retroperitoneal adipocyte areas were found in the resistance training high-fat group than in the sedentary high-fat group. In the swimming high-fat group, the tumor necrosis factor-alpha expression was lower and the epididymal and retroperitoneal adipocyte areas were smaller compared with the sedentary high-fat group. CONCLUSION: The results showed that both exercise modalities improved the lipid profile, adiposity and obesity-associated inflammation in rats, suggesting their use as an alternative to control the deleterious effects of a high-fat diet in humans.

The effects of exercise modalities on adiposity in obese rats.

OBJECTIVE: The aim of the present study was to evaluate the effect of both swimming and resistance training on tumor necrosis factor-alpha and interleukin-10 expression, adipocyte area and lipid profiles in rats fed a high-fat diet. METHODS: The study was conducted over an eight-week period on Wistar adult rats, who were divided into six groups as follows (n = 10 per group): sedentary chow diet, sedentary high-fat diet, swimming plus chow diet, swimming plus high-fat diet, resistance training plus chow diet, and resistance training plus high-fat diet. Rats in the resistance training groups climbed a vertical ladder with weights on their tails once every three days. The swimming groups swam for 60 minutes/day, five days/week. RESULTS: The high-fat diet groups had higher body weights, a greater amount of adipose tissue, and higher tumor necrosis factor-alpha expression in the visceral adipose tissue. Furthermore, the high-fat diet promoted a negative change in the lipid profile. In the resistance training high-fat group, the tumor necrosis factor-alpha expression was lower than that in the swimming high-fat and sedentary high-fat groups. Moreover, smaller visceral and retroperitoneal adipocyte areas were found in the resistance training high-fat group than in the sedentary high-fat group. In the swimming high-fat group, the tumor necrosis factor-alpha expression was lower and the epididymal and retroperitoneal adipocyte areas were smaller compared with the sedentary high-fat group. CONCLUSION: The results showed that both exercise modalities improved the lipid profile, adiposity and obesity-associated inflammation in rats, suggesting their use as an alternative to control the deleterious effects of a high-fat diet in humans.