Resumo
Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa- ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of 6.9, 7.6, 7.1, 6.9, 6.7, and 7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 g / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 g / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.
Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa- [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de 6,9, 7,6, 7,1, 6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 g / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 g / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.
Resumo
Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.
Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.
Assuntos
Osteoclastos , Ligante RANK , Diferenciação Celular , Simulação de Acoplamento Molecular , Resveratrol/farmacologiaResumo
Abstract Cisplatin (CP) is a commonly used, powerful antineoplastic drug, having numerous side effects. Casticin (CAS) is considered as a free radical scavenger and a potent antioxidant. The present research was planned to assess the curative potential of CAS on CP persuaded renal injury in male albino rats. Twenty four male albino rats were distributed into four equal groups. Group-1 was considered as a control group. Animals of Group-2 were injected with 5mg/kg of CP intraperitoneally. Group-3 was co-treated with CAS (50mg/kg) orally and injection of CP (5mg/kg). Group-4 was treated with CAS (50mg/kg) orally throughout the experiment. CP administration substantially reduced the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) content while increased thiobarbituric acid reactive substances (TBARS), and hydrogen peroxide (H2O2) levels. Urea, urinary creatinine, urobilinogen, urinary proteins, kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) levels were substantially increased. In contrast, albumin and creatinine clearance was significantly reduced in CP treated group. The results demonstrated that CP significantly increased the inflammation indicators including nuclear factor kappa-B (NF-B), tumor necrosis factor- (TNF-), Interleukin-1 (IL-1), Interleukin-6 (IL-6) levels and cyclooxygenase-2 (COX-2) activity and histopathological damages. However, the administration of CAS displayed a palliative effect against CP-generated renal toxicity and recovered all parameters by bringing them to a normal level. These results revealed that the CAS is an effective compound having the curative potential to counter the CP-induced renal damage.
Resumo A cisplatina (CP) é uma droga antineoplásica poderosa, comumente usada, com vários efeitos colaterais. Casticin (CAS) é considerado um eliminador de radicais livres e um potente antioxidante. A presente pesquisa foi planejada para avaliar o potencial curativo da CAS em lesão renal induzida por PC em ratos albinos machos. Vinte e quatro ratos albinos machos foram distribuídos em quatro grupos iguais. O Grupo 1 foi considerado grupo controle. Os animais do Grupo 2 foram injetados com 5 mg / kg de PB por via intraperitoneal. O Grupo 3 foi cotratado com CAS (50 mg / kg) por via oral e injeção de CP (5 mg / kg). O Grupo 4 foi tratado com CAS (50 mg / kg) por via oral durante todo o experimento. A administração de CP reduziu substancialmente as atividades de catalase (CAT), superóxido dismutase (SOD), peroxidase (POD), glutationa S-transferase (GST), glutationa redutase (GSR), glutationa (GSH), enquanto aumentou as substâncias reativas ao ácido tiobarbitúrico (TBARS) e níveis de peróxido de hidrogênio (H2O2). Os níveis de ureia, creatinina urinária, urobilinogênio, proteínas urinárias, molécula 1 de lesão renal (KIM-1) e lipocalina associada à gelatinase de neutrófilos (NGAL) aumentaram substancialmente. Em contraste, a albumina e a depuração da creatinina foram significativamente reduzidas no grupo tratado com PC. Os resultados demonstraram que a CP aumentou significativamente os indicadores de inflamação, incluindo fator nuclear kappa-B (NF-B), fator de necrose tumoral- (TNF-), interleucina-1 (IL-1), interleucina-6 (IL-6) níveis e atividade da ciclooxigenase-2 (COX-2) e danos histopatológicos. No entanto, a administração de CAS apresentou um efeito paliativo contra a toxicidade renal gerada por CP e recuperou todos os parâmetros, trazendo-os a um nível normal. Estes resultados revelaram que o CAS é um composto eficaz com potencial curativo para combater o dano renal induzido por CP.
Resumo
By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.
Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.
Assuntos
Humanos , Osteoporose/tratamento farmacológico , Resveratrol/farmacologia , Microscopia de FluorescênciaResumo
By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.(AU)
Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.(AU)
Assuntos
Humanos , Osteoporose/tratamento farmacológico , Resveratrol/farmacologia , Microscopia de FluorescênciaResumo
Cisplatin (CP) is a commonly used, powerful antineoplastic drug, having numerous side effects. Casticin (CAS) is considered as a free radical scavenger and a potent antioxidant. The present research was planned to assess the curative potential of CAS on CP persuaded renal injury in male albino rats. Twenty four male albino rats were distributed into four equal groups. Group-1 was considered as a control group. Animals of Group-2 were injected with 5mg/kg of CP intraperitoneally. Group-3 was co-treated with CAS (50mg/kg) orally and injection of CP (5mg/kg). Group-4 was treated with CAS (50mg/kg) orally throughout the experiment. CP administration substantially reduced the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) content while increased thiobarbituric acid reactive substances (TBARS), and hydrogen peroxide (H2O2) levels. Urea, urinary creatinine, urobilinogen, urinary proteins, kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) levels were substantially increased. In contrast, albumin and creatinine clearance was significantly reduced in CP treated group. The results demonstrated that CP significantly increased the inflammation indicators including nuclear factor kappa-B (NF-κB), tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), Interleukin-6 (IL-6) levels and cyclooxygenase-2 (COX-2) activity and histopathological damages. However, the administration of CAS displayed a palliative effect against CP-generated renal toxicity and recovered all parameters by bringing them to a normal level. These results revealed that the CAS is an effective compound having the curative potential to counter the CP-induced renal damage.
A cisplatina (CP) é uma droga antineoplásica poderosa, comumente usada, com vários efeitos colaterais. Casticin (CAS) é considerado um eliminador de radicais livres e um potente antioxidante. A presente pesquisa foi planejada para avaliar o potencial curativo da CAS em lesão renal induzida por PC em ratos albinos machos. Vinte e quatro ratos albinos machos foram distribuídos em quatro grupos iguais. O Grupo 1 foi considerado grupo controle. Os animais do Grupo 2 foram injetados com 5 mg / kg de PB por via intraperitoneal. O Grupo 3 foi cotratado com CAS (50 mg / kg) por via oral e injeção de CP (5 mg / kg). O Grupo 4 foi tratado com CAS (50 mg / kg) por via oral durante todo o experimento. A administração de CP reduziu substancialmente as atividades de catalase (CAT), superóxido dismutase (SOD), peroxidase (POD), glutationa S-transferase (GST), glutationa redutase (GSR), glutationa (GSH), enquanto aumentou as substâncias reativas ao ácido tiobarbitúrico (TBARS) e níveis de peróxido de hidrogênio (H2O2). Os níveis de ureia, creatinina urinária, urobilinogênio, proteínas urinárias, molécula 1 de lesão renal (KIM-1) e lipocalina associada à gelatinase de neutrófilos (NGAL) aumentaram substancialmente. Em contraste, a albumina e a depuração da creatinina foram significativamente reduzidas no grupo tratado com PC. Os resultados demonstraram que a CP aumentou significativamente os indicadores de inflamação, incluindo fator nuclear kappa-B (NF-κB), fator de necrose tumoral-α (TNF-α), interleucina-1β (IL-1β), interleucina-6 (IL-6) níveis e atividade da ciclooxigenase-2 (COX-2) e danos histopatológicos. No entanto, a administração de CAS apresentou um efeito paliativo contra a toxicidade renal gerada por CP e recuperou todos os parâmetros, trazendo-os a um nível normal. Estes resultados revelaram que o CAS é um composto eficaz com potencial curativo para combater o dano renal induzido por CP.
Assuntos
Masculino , Animais , Ratos , Antineoplásicos/administração & dosagem , Antineoplásicos/efeitos adversos , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Rim/lesões , Sequestradores de Radicais Livres/administração & dosagem , Sequestradores de Radicais Livres/farmacologia , Ratos EndogâmicosResumo
Cisplatin (CP) is a commonly used, powerful antineoplastic drug, having numerous side effects. Casticin (CAS) is considered as a free radical scavenger and a potent antioxidant. The present research was planned to assess the curative potential of CAS on CP persuaded renal injury in male albino rats. Twenty four male albino rats were distributed into four equal groups. Group-1 was considered as a control group. Animals of Group-2 were injected with 5mg/kg of CP intraperitoneally. Group-3 was co-treated with CAS (50mg/kg) orally and injection of CP (5mg/kg). Group-4 was treated with CAS (50mg/kg) orally throughout the experiment. CP administration substantially reduced the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) content while increased thiobarbituric acid reactive substances (TBARS), and hydrogen peroxide (H2O2) levels. Urea, urinary creatinine, urobilinogen, urinary proteins, kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) levels were substantially increased. In contrast, albumin and creatinine clearance was significantly reduced in CP treated group. The results demonstrated that CP significantly increased the inflammation indicators including nuclear factor kappa-B (NF-κB), tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), Interleukin-6 (IL-6) levels and cyclooxygenase-2 (COX-2) activity and histopathological damages. However, the administration of CAS displayed a palliative effect against CP-generated renal toxicity and recovered all parameters by bringing them to a normal level. These results revealed that the CAS is an effective compound having the curative potential to counter the CP-induced renal damage.(AU)
A cisplatina (CP) é uma droga antineoplásica poderosa, comumente usada, com vários efeitos colaterais. Casticin (CAS) é considerado um eliminador de radicais livres e um potente antioxidante. A presente pesquisa foi planejada para avaliar o potencial curativo da CAS em lesão renal induzida por PC em ratos albinos machos. Vinte e quatro ratos albinos machos foram distribuídos em quatro grupos iguais. O Grupo 1 foi considerado grupo controle. Os animais do Grupo 2 foram injetados com 5 mg / kg de PB por via intraperitoneal. O Grupo 3 foi cotratado com CAS (50 mg / kg) por via oral e injeção de CP (5 mg / kg). O Grupo 4 foi tratado com CAS (50 mg / kg) por via oral durante todo o experimento. A administração de CP reduziu substancialmente as atividades de catalase (CAT), superóxido dismutase (SOD), peroxidase (POD), glutationa S-transferase (GST), glutationa redutase (GSR), glutationa (GSH), enquanto aumentou as substâncias reativas ao ácido tiobarbitúrico (TBARS) e níveis de peróxido de hidrogênio (H2O2). Os níveis de ureia, creatinina urinária, urobilinogênio, proteínas urinárias, molécula 1 de lesão renal (KIM-1) e lipocalina associada à gelatinase de neutrófilos (NGAL) aumentaram substancialmente. Em contraste, a albumina e a depuração da creatinina foram significativamente reduzidas no grupo tratado com PC. Os resultados demonstraram que a CP aumentou significativamente os indicadores de inflamação, incluindo fator nuclear kappa-B (NF-κB), fator de necrose tumoral-α (TNF-α), interleucina-1β (IL-1β), interleucina-6 (IL-6) níveis e atividade da ciclooxigenase-2 (COX-2) e danos histopatológicos. No entanto, a administração de CAS apresentou um efeito paliativo contra a toxicidade renal gerada por CP e recuperou todos os parâmetros, trazendo-os a um nível normal. Estes resultados revelaram que o CAS é um composto eficaz com potencial curativo para combater o dano renal induzido por CP.(AU)
Assuntos
Animais , Masculino , Ratos , Rim/lesões , Antineoplásicos/administração & dosagem , Antineoplásicos/efeitos adversos , Sequestradores de Radicais Livres/administração & dosagem , Sequestradores de Radicais Livres/farmacologia , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Ratos EndogâmicosResumo
ABSTRACT Purpose To demonstrate the effect of IL-33 on the macrophage pyroptosis in mice with sepsis through the NF-kB/p38 MAPK signal pathway. Methods In total, 24 C57BL/6 mice were divided into the sham operation group (sham) and the cecal ligation and puncture group (CLP). After CLP, 24 IL-33-/- mice were divided into the IL-33-/- group and the IL-33-/- intervention group. The latter group was intraperitoneally injected with IL-33. Mouse mortality was observed after CLP. Macrophage apoptosis in peritoneal lavage fluid was detected by flow cytometry. Serum inflammatory factor level was detected by ELISA. Apoptotic protein expression and NF-κB/p38 MAKP signaling pathway protein expression were detected by qRT-PCR and Western blot. Results Knocking out IL-33 significantly reduced the mortality of CLP mice, as well as the mRNA expression of IL-33 and the levels of serum inflammatory factors, including IL-33, IL-1β, and IL-18. It also reduced the rate of macrophage apoptosis and the expression of the apoptotic protein caspase-1 p10; increased the expression of IκBα; and reduced the protein expression of NF-κB and p38 MAPK. These effects were reversed after exogenous injection of IL-33. Conclusions IL-33 can increase the level of macrophage pyroptosis in mice with sepsis (by activating the NF-kB/p38MAPK signal pathway) and the mortality of these mice.
Assuntos
Animais , Camundongos , NF-kappa B/metabolismo , Sepse , Transdução de Sinais , Fator de Necrose Tumoral alfa , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Interleucina-33 , Piroptose , Macrófagos/metabolismo , Camundongos Endogâmicos C57BLResumo
Chicken coccidiosis is a parasitic disease caused by one or more species of Eimeria infection with severe consequences. The NF-B signaling pathway and TGF-4 play an important role in the inflammation and immune response caused by coccidiosis infection. This study was designed to investigate the changes of NF-B signaling pathway and the effects of TGF-4 silencing on the expression of NF-B signaling pathway in chickens intestinal epithelial cells (IECs) after infecting with E. tenella sporozoites. TGF-4 small interfering RNA (TGF-4 siRNA) sequences were transfected into chicken IECs for reducing TGF-4 expression. The results showed that compared with uninfected control group (Con.), the proinflammatory factors of NF-B, IL-6, IL-2, IL-1 increased significantly in the E. tenella infected group (ET) (p 0.05). In comparison with the IECs in the ET, the expression level of NF-B, IL-6, IL-2, IL-1 dropped significantly in the group infected both by E. tenella and TGF-4 siRNA vector (ET+siRNA) (p 0.05). The results of this experiment indicate that NF-B signaling pathway is closely correlated with TGF-4 in IECs infected with coccidia sporozoites. TGF-4 plays an important role in regulating the immune function of coccidia-infected chicken epithelial cells through NF-B signaling pathway and preventing immune-mediated pathological changes.(AU)
Assuntos
Animais , Feminino , NF-kappa B/análise , NF-kappa B/classificação , Fator de Crescimento Transformador beta/análise , Células Epiteliais , Eimeria tenella/patogenicidade , CoccidioseResumo
Chicken coccidiosis is a parasitic disease caused by one or more species of Eimeria infection with severe consequences. The NF-B signaling pathway and TGF-4 play an important role in the inflammation and immune response caused by coccidiosis infection. This study was designed to investigate the changes of NF-B signaling pathway and the effects of TGF-4 silencing on the expression of NF-B signaling pathway in chickens intestinal epithelial cells (IECs) after infecting with E. tenella sporozoites. TGF-4 small interfering RNA (TGF-4 siRNA) sequences were transfected into chicken IECs for reducing TGF-4 expression. The results showed that compared with uninfected control group (Con.), the proinflammatory factors of NF-B, IL-6, IL-2, IL-1 increased significantly in the E. tenella infected group (ET) (p 0.05). In comparison with the IECs in the ET, the expression level of NF-B, IL-6, IL-2, IL-1 dropped significantly in the group infected both by E. tenella and TGF-4 siRNA vector (ET+siRNA) (p 0.05). The results of this experiment indicate that NF-B signaling pathway is closely correlated with TGF-4 in IECs infected with coccidia sporozoites. TGF-4 plays an important role in regulating the immune function of coccidia-infected chicken epithelial cells through NF-B signaling pathway and preventing immune-mediated pathological changes.
Assuntos
Feminino , Animais , Coccidiose , Células Epiteliais , Eimeria tenella/patogenicidade , Fator de Crescimento Transformador beta/análise , NF-kappa B/análise , NF-kappa B/classificaçãoResumo
Purpose. To evaluate the neuroprotective effect of L-alanyl-glutamine in a gerbil model of brain ischemia-reperfusion injury based on immunohistochemical quantification of pro-inflammatory and cell activation biomarkers (TNF-α, NF-κB, IL-6 and HO-1).. Methods. Male gerbils weighing 100-180 g were pretreated with either 0.75 g/kg L-Ala-Gln (n=18) or 2.0 mL saline (n=18) administered i.v. 30 minutes before the bilateral ligation of the common carotid artery during 15 min and then the ligation was removed. Under anesthesia with urethane, brain tissue was harvested at 0 min (T0), 30 min (T30) and 60 min (T60) after reperfusion. The tissue was embedded in 10% formalin overnight and 4-μm sections were prepared for immunostaining with monoclonal antibodies. Immunostained cells were counted by optical microscopy. The statistical analysis used mean values based on 4 sections.. Results. The pretreatment with L-Ala-Gln animal group 1 demonstrated significantly lower levels of TNF-α, NF-κB and IL-6. On the other hand, the levels of HO-1 were significantly higher, suggesting a protective role in model of brain ischemia-reperfusion injury.. Conclusion. These findings suggest a protective effect of L-Ala-Gln by decreasing levels of TNF-alpha, IL-6 and NF-κB and Increasing levels of HO-1.(AU)
Assuntos
Animais , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/classificação , NF-kappa B/análise , Interleucina-6/análise , Isquemia , GerbillinaeResumo
Purpose:To explore the effect of milk fat globule-epidermal growth factor 8 (MFG-E8) on sepsis-induced acute kidney injury (SAKI).Methods:Male C57BL/6 mice were randomized to control, sham, CLP, CLP+PBS, and CLP+rmMFG-E8 groups. SAKI was induced by cecal ligation and puncture (CLP). Recombinant mouse MFG-E8 (rmMFG-E8) (20 μg/kg) or PBS (vehicle) was administered intraperitoneally. Blood, urine and renal tissue were collected at 24 h after CLP. Blood samples were tested for serum kidney injury biomarker and cytokines. Urine samples were collected to detect KIM-1, and NGAL. Real-time PCR was tested for Bax and Bcl-2. TUNEL staining was used to determine renal apoptosis. Western blot was used to detect the expression of Bax, Bcl-2, and proteins in the NF-κB pathway.Results:MFG-E8 alleviated SAKI by decreasing serum Cre, BUN, urine KIM-1 and NGAL and by mitigating renal pathological changes significant (p < 0.05). IL-1β, IL-6, TNF-α were significantly inhibited by MFG-E8 (p < 0.05). Apoptosis induced by SAKI was markedly suppressed by MFG-E8. Finally, MFG-E8 attenuated the activation of the NF-𝜅B signaling pathway in SAKI.Conclusion:MFG-E8 has beneficial effects on SAKI, which may be achieved by inhibiting the NF-κB pathway.(AU)
Assuntos
Animais , Ratos , Fator de Crescimento Epidérmico/análise , Injúria Renal Aguda/veterinária , Sepse , NF-kappa B/antagonistas & inibidores , Inflamação , ApoptoseResumo
Avian leukosis virus subgroup J (ALV-J), a member of the retroviridae family, can infect both broilers and layers and induce a spectrum of different neoplasms, resulting in serious economic losses in poultry production. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, has demonstrated remarkable anti-inflammatory and cancer chemopreventive effects in many animal tumor bioassays, cell culture systems and epidemiological studies. To assess the antiviral effects of EGCG on ALV-J-induced cell apoptosis in vitro, DF-1 cells were treated with different EGCG concentrations (0, 5, 10, 20 and 40 µg/mL), and their antiviral effects were examined at different time points (0, 24, 48, 72 and 96 h) using a variety of assays. EGCG alleviated the ALV-J-induced apoptosis in a dose-dependent manner. Because high concentrations (20 and 40 µg/mL) inhibited DF-1 cell growth, and low concentration (5 µg/mL) did not suppress the ALV-J virus, 10 µg/mL was the most appropriate concentration. After 96 h of incubation, 10 µg/mL EGCG improved the ALV-J-triggered suppression of the nuclear transcription factor system by enhancing cytoplasmic NF-B p50/p65 expression and inhibiting nuclear NF-B p50/p65 expression, resulting in decreased cell apoptosis. These results demonstrated that EGCG inhibited ALV-J-induced apoptosis in DF-1 cells in a dose-dependent manner via the NF-B signaling pathway, and that 10 µg/mL EGCG is the optimal concentration, which may be useful for therapeutic drug design.(AU)
Assuntos
Animais , Galinhas/virologia , Apoptose , Galato de Propila/química , NF-kappa B/análise , Vírus da Leucose AviáriaResumo
Avian leukosis virus subgroup J (ALV-J), a member of the retroviridae family, can infect both broilers and layers and induce a spectrum of different neoplasms, resulting in serious economic losses in poultry production. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, has demonstrated remarkable anti-inflammatory and cancer chemopreventive effects in many animal tumor bioassays, cell culture systems and epidemiological studies. To assess the antiviral effects of EGCG on ALV-J-induced cell apoptosis in vitro, DF-1 cells were treated with different EGCG concentrations (0, 5, 10, 20 and 40 µg/mL), and their antiviral effects were examined at different time points (0, 24, 48, 72 and 96 h) using a variety of assays. EGCG alleviated the ALV-J-induced apoptosis in a dose-dependent manner. Because high concentrations (20 and 40 µg/mL) inhibited DF-1 cell growth, and low concentration (5 µg/mL) did not suppress the ALV-J virus, 10 µg/mL was the most appropriate concentration. After 96 h of incubation, 10 µg/mL EGCG improved the ALV-J-triggered suppression of the nuclear transcription factor system by enhancing cytoplasmic NF-B p50/p65 expression and inhibiting nuclear NF-B p50/p65 expression, resulting in decreased cell apoptosis. These results demonstrated that EGCG inhibited ALV-J-induced apoptosis in DF-1 cells in a dose-dependent manner via the NF-B signaling pathway, and that 10 µg/mL EGCG is the optimal concentration, which may be useful for therapeutic drug design.
Assuntos
Animais , Apoptose , Galato de Propila/química , Galinhas/virologia , NF-kappa B/análise , Vírus da Leucose AviáriaResumo
Purpose To examine the therapeutic effect of external adenosine on an acetic acid-induced acute ulcerative colitis model in rats. Methods Thirty male mature rats were divided into three groups as control, acute colitis (AC) and AC+adenosine group (AC+AD). AC was induced by rectal administration of 4% acetic acid (AA). 5mg/kg/day adenosine was performed i.p for 4 weeks to AC+AD group. Rectum and colon were excised for microscopic and histopathological histopathologic evaluations, and immunohistochemical analysis of nuclear factor kappa B (NF-kB). Blood samples were collected for biochemical detection of TNF-α, Pentraxin-3 and malondialdehyde (MDA) levels. Results AC group had generalized hyperemia and hemorrhage with increased macroscopic and histopathological scores compared with control (P <0.0001) while adenosine treatment decreased these scores significantly (P <0.001), with reduced distribution of disrupted epithelium, leukocyte infiltrates, and focal hemorrhage. AC group showed significantly increased immunoexpression of NF-kB in rectum, plasma and tissue levels of TNF-α, plasma Pentraxin-3 and MDA levels (P <0.0001) while adenosine reduced these levels (P < 0.05). Conclusion Adenosine appears to promote healing of colon and rectum exposed to AA-induced AC, suggesting a boosting effect of adenosine on the intestinal immune system to cure ulcerative colitis.(AU)
Assuntos
Animais , Masculino , Ratos , Colite Ulcerativa/tratamento farmacológico , Adenosina/uso terapêutico , NF-kappa B/antagonistas & inibidores , Modelos AnimaisResumo
Purpose: To investigate the place of the transcription factor nuclear kappa B (NF-kB), which is a marker of chronic inflammation, in the etiology of the ovarian carcinoma. Methods: NFkB analysis with the immunohistochemical method has been performed. To evaluate immunohistochemical NF-kB expression in the ovarian tissue, the H-score method. H-score = Pi (i+1), where Pi is the percentage of stained cells in each intensity category (0-100%) and i is the intensity indicating weak (i=1), moderate (i=2) or strong staining (i=3). Results: It has been seen that, the mean H score is statistically significantly higher in the patient group with serous and musinous adenocarcinoma diagnosis than the two other patient groups (p 0.005). Conclusions: Factor nuclear kappa B is an important mediator that acts in the chronic inflammation. The highest expression rates are determined by the immunohistochemical method in the ovarian cancer group.(AU)
Assuntos
Humanos , NF-kappa B/análise , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/etiologia , Imuno-Histoquímica/métodosResumo
Purpose: To evaluate the role of CX3CL1 and NF-B in the lumbar disc herniation induced neuropathic pain. Methods: After LDH induced by implantation of autologous nucleus pulposus (NP) on the left L5 nerve root was established, mechanical thresholds and thermal hyperalgesia were tested at relevant time points during an observation period of 28 days. Expression of CX3CL1 and NF-Bin the dorsal root ganglion (DRG) were performed by using Western blotting and RT-PCR. Results: Implantation of autologous nucleus pulposus (NP) induced neuropathic pain, associated with increased mRNA and protein expression of CX3CL1 in the DRG. Moreover, intrathecal injection of neutralizing antibody against CX3CL1 could attenuates LDH-induced persistent pain hypersensitivity. Interestingly, NF-B activation in the DRGs were found in LDH-induced neuropathic pain. Furthermore, NF-B downregulation by p65 inhibitor PDTC markedly alleviated LDH-induced mechanical allodynia and thermal hyperalgesia in rat. Importantly, CX3CL1 neutralizing antibody (10 g/10 l, i.t.) reduces p-p65 protein level in DRG Conclusions: CX3XL1 could regulate LDH-induced neuropathic pain through NF-B pathway. Targeting CX3CL1 and NF-B may represent a potential treatment for neuropathic pain caused by LDH.(AU)
Assuntos
Animais , Masculino , Adulto , Ratos , NF-kappa B/análise , Quimiocina CX3CL1/análise , Neuralgia/induzido quimicamente , Gânglios Espinais/patologia , Modelos Animais , Ratos Sprague-DawleyResumo
This study aims to investigate whether resveratrol (RES) could inhibit NF-κB activity and protect in vitro cultured chicken lymphocytes from apoptosis induced by continuous LPS stimulation. Blood lymphocytes of chickens were collected and cultured. LPS was added to the samples of treatment group, while the same volume of normal saline (NS) was supplemented to those of the control group. In the treatment groups, four different concentrations of RES (0, 20, 40, and 80 µg/ml) were admixed. Then, the following indicators were tested: lymphocyte apoptotic rate, lymphocyte viability, NF-κB activity, TNF-a level, reactive oxygen species (ROS), and expression of apoptosis-related proteins (Fas/FasL and Caspase-8). The results indicated that the application of different concentrations of RES could significantly increase the viability of lymphocytes and decrease the apoptotic rate (p<0.05) after continuous stimulations by LPS. The activity of NF-κB levels of TNF-a and ROS in the RES treatment subgroups was significantly decreased in comparison with that in the RES-blank subgroup (p<0.05). The cells in the treatment group that had been treated with 20, 40 and 80 μl/ml of RES exhibited a significantly lower Fas and Caspase-8 expression level than that in the RES-blank subgroup (p<0.05). These findings revealed that RES could substantially diminish the concentrations of ROS and TNF-a, down-regulate NF-κB activity, decrease the expression of Fas and Caspase-8, stimulate the activity of peripheral lymphocytes, and lower the rate of their apoptosis, induced by continuous LPS stimulation in chicken lymphocyte cultures.(AU)
Assuntos
Animais , Recém-Nascido , NF-kappa B/antagonistas & inibidores , Apoptose , Galinhas/sangue , Galinhas/imunologia , Polifenóis/efeitos adversos , Polifenóis/uso terapêutico , Lipopolissacarídeos , Linfócitos/patologiaResumo
This study aims to investigate whether resveratrol (RES) could inhibit NF-κB activity and protect in vitro cultured chicken lymphocytes from apoptosis induced by continuous LPS stimulation. Blood lymphocytes of chickens were collected and cultured. LPS was added to the samples of treatment group, while the same volume of normal saline (NS) was supplemented to those of the control group. In the treatment groups, four different concentrations of RES (0, 20, 40, and 80 µg/ml) were admixed. Then, the following indicators were tested: lymphocyte apoptotic rate, lymphocyte viability, NF-κB activity, TNF-a level, reactive oxygen species (ROS), and expression of apoptosis-related proteins (Fas/FasL and Caspase-8). The results indicated that the application of different concentrations of RES could significantly increase the viability of lymphocytes and decrease the apoptotic rate (p<0.05) after continuous stimulations by LPS. The activity of NF-κB levels of TNF-a and ROS in the RES treatment subgroups was significantly decreased in comparison with that in the RES-blank subgroup (p<0.05). The cells in the treatment group that had been treated with 20, 40 and 80 μl/ml of RES exhibited a significantly lower Fas and Caspase-8 expression level than that in the RES-blank subgroup (p<0.05). These findings revealed that RES could substantially diminish the concentrations of ROS and TNF-a, down-regulate NF-κB activity, decrease the expression of Fas and Caspase-8, stimulate the activity of peripheral lymphocytes, and lower the rate of their apoptosis, induced by continuous LPS stimulation in chicken lymphocyte cultures.
Assuntos
Animais , Recém-Nascido , Apoptose , Galinhas/imunologia , Galinhas/sangue , NF-kappa B/antagonistas & inibidores , Polifenóis/efeitos adversos , Polifenóis/uso terapêutico , Linfócitos/patologia , LipopolissacarídeosResumo
Purpose: To investigate the effects of baicalin on inflammatory reaction, oxidative stress and protein kinase D1 (PKD1) and nuclear factor-kappa B (NF-B) protein expressions in severe acute pancreatitis (SAP) rats. Methods: Sixty rats were divided into sham operation, model, and low-, medium- and high-dose baicalin group. SAP model was established in later 4 groups. The later 3 groups were injected with 0.1, 0.2 and 0.4 ml/100 g 5% baicalin injection, respectively. At 12 h, the serum SAP related indexes and inflammatory factors, peripheral blood CD3 and T cell percentages, wet/dry ratio and pancreas ascites volume, oxidative stress indexes and PKD1 and NF-B protein expressions in pancreatic tissue were determined. Results: Compared with model group, in high-dose baicalin group the wet/dry ratio and ascites volume, serum amylase level, phospholipase A2 activity, TNF-, IL-1 and IL-6 levels, and pancreatic malondialdehyde level and PKD1 and NF-B protein expression were significantly decreased (P 0.05), and peripheral blood CD3 and T cell percentages and pancreatic superoxide dismutase and glutathione peroxidase levels were significantly increased (P 0.05). Conclusion: Baicalin can resist the inflammatory reaction and oxidative stress, and down-regulate protein kinase D1 and nuclear factor-kappa B protein expressions, thus exerting the protective effects on severe acute pancreatitis in rats.(AU)