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SUMMARY: Gastrin plays a vital role in the development and progression of gastric cancer (GC). Its expression is up-regulated in GC tissues and several GC cell lines. Yet, the underlying mechanism remains to be investigated. Here, we aim to investigate the role and mechanism of gastrin in GC proliferation. Gastrin-overexpressing GC cell model was constructed using SGC7901 cells. Then the differentially expressed proteins were identified by iTRAQ analysis. Next, we use flow cytometry and immunofluorescence to study the effect of gastrin on the mitochondrial potential and mitochondria-derived ROS production. Finally, we studied the underlying mechanism of gastrin regulating mitochondrial function using Co-IP, mass spectrometry and immunofluorescence. Overexpression of gastrin promoted GC cell proliferation in vitro and in vivo. A total of 173 proteins were expressed differently between the controls and gastrin- overexpression cells and most of these proteins were involved in tumorigenesis and cell proliferation. Among them, Cox17, Cox5B and ATP5J that were all localized to the mitochondrial respiratory chain were down-regulated in gastrin-overexpression cells. Furthermore, gastrin overexpression led to mitochondrial potential decrease and mitochondria-derived ROS increase. Additionally, gastrin-induced ROS generation resulted in the inhibition of cell apoptosis via activating NF-kB, inhibiting Bax expression and promoting Bcl-2 expression. Finally, we found gastrin interacted with mitochondrial membrane protein Annexin A2 using Co-IP and mass spectrometry. Overexpr ession of gastrin inhibits GC cell apoptosis by inducing mitochondrial dysfunction through interacting with mitochondrial protein Annexin A2, then up-regulating ROS production to activate NF-kB and further leading to Bax/Bcl-2 ratio decrease.
La gastrina juega un papel vital en el desarrollo y progresión del cáncer gástrico (CG). Su expresión está regulada al alza en tejidos de CG y en varias líneas celulares de CG. Sin embargo, el mecanismo subyacente aun no se ha investigado. El objetivo de este estudio fue investigar el papel y el mecanismo de la gastrina en la proliferación de CG. El modelo de células CG que sobre expresan gastrina se construyó usando células SGC7901. Luego, las proteínas expresadas diferencialmente se identificaron mediante análisis iTRAQ. A continuación, utilizamos la citometría de flujo y la inmunofluorescencia para estudiar el efecto de la gastrina en el potencial mitocondrial y la producción de ROS derivada de las mitocondrias. Finalmente, estudiamos el mecanismo subyacente de la gastrina que regula la función mitocondrial utilizando Co-IP, espectrometría de masas e inmunofluorescencia. La sobreexpresión de gastrina promovió la proliferación de células CG in vitro e in vivo. Un total de 173 proteínas se expresaron de manera diferente entre los controles y las células con sobreexpresión de gastrina y la mayoría de estas proteínas estaban implicadas en la tumorigenesis y la proliferación celular. Entre estas, Cox17, Cox5B y ATP5J, todas localizadas en la cadena respiratoria mitocondrial, estaban reguladas a la baja en las células con sobreexpresión de gastrina. Además, la sobreexpresión de gastrina provocó una disminución del potencial mitocondrial y un aumento de las ROS derivadas de las mitocondrias. Por otra parte, la generación de ROS inducida por gastrina resultó en la inhibición de la apoptosis celular mediante la activación de NF-kB, inhibiendo la expresión de Bax y promoviendo la expresión de Bcl-2. Finalmente, encontramos que la gastrina interactuaba con la proteína de membrana mitocondrial Anexina A2 usando Co-IP y espectrometría de masas. La sobreexpresión de gastrina inhibe la apoptosis de las células CG al inducir la disfunción mitocondrial a través de la interacción con la proteína mitocondrial Anexina A2, luego regula el aumento de la producción de ROS para activar NF-kB y conduce aún más a la disminución de la relación Bax/Bcl-2.
Subject(s)
Animals , Mice , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Gastrins/metabolism , Annexin A2/metabolism , Mitochondria/pathology , Mass Spectrometry , NF-kappa B , Fluorescent Antibody Technique , Reactive Oxygen Species , Apoptosis , Cell Line, Tumor , Immunoprecipitation , Cell Proliferation , Carcinogenesis , Flow CytometryABSTRACT
SUMMARY: Paracetamol (known as acetaminophen, or APAP) poisoning causes acute liver damage that can lead to organ failure and death. We sought to determine that APAP overdose can augment tumor necrosis factor-alpha (TNF-α)/ nuclear factor kappa B (NF-kB)/induced nitic oxide synthase (iNOS) axis-mediated hepatotoxicity in rats, and the anti-inflammatory polyphenolic compounds, quercetin (QUR) plus resveratrol (RES) can ameliorate these parameters. Therefore, we induced acute hepatotoxicity in rats using APAP overdose (2 g/kg, orally) and the protective group of rats were treated with 50 mg/kg QUR plus 30 mg/kg RES for one week before APAP ingestion. Animals were killed at day 8. APAP poisoning caused the induction of hepatic tissue levels of TNF-α, NF-kB, and iNOS, which were significantly (p<0.05) decreased by QUR+RES. QUR+RES, also inhibited liver injury biomarkers, alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Additionally, a link between liver injury and TNF-α /NF-kB / iNOS axis mediated hepatotoxicity was observed. Thus, the presented data backing the conclusion that intoxication by paracetamol increases TNF-α / NF-kB / iNOS axis -mediated hepatotoxicity, and is protected by a combination of quercetin and resveratrol.
El envenenamiento por paracetamol (conocido como acetaminofeno o APAP) causa daño hepático agudo que puede provocar una insuficiencia orgánica y la muerte. El objetivo de este trabajo fue determinar si la sobredosis de APAP puede aumentar la hepatotoxicidad mediada por el eje del factor de necrosis tumoral alfa (TNF-α)/factor nuclear kappa B (NF-kB)/óxido nítico sintasa inducida (iNOS) en ratas, y si el polifenólico antiinflamatorio compuesto por quercetina (QUR) más resveratrol (RES) pueden mejorar estos parámetros. Por lo tanto, inducimos hepatotoxicidad aguda en ratas usando una sobredosis de APAP (2 g/kg, por vía oral). El grupo protector de ratas se trató con 50 mg/ kg de QUR más 30 mg/kg de RES durante una semana antes de la ingestión de APAP. Los animales se sacrificaron el día 8. El envenenamiento con APAP en el tejido hepático provocó la inducción de niveles de TNF-α, NF-kB e iNOS, que se redujeron significativamente (p<0,05) con QUR+RES. QUR+RES, también inhibió los biomarcadores de daño hepático, la alanina aminotransferasa (ALT) y el aspartato aminotransferasa (AST). Además, se observó una relación entre la lesión hepática y la hepatotoxicidad mediada por el eje TNF-α /NF-kB/iNOS. Por lo tanto, los datos presentados respaldan la conclusión de que la intoxicación por paracetamol aumenta la hepatotoxicidad mediada por el eje TNF-α /NF-kB / iNOS, y está protegida por una combinación de quercetina y resveratrol.
Subject(s)
Animals , Rats , Quercetin/administration & dosage , Chemical and Drug Induced Liver Injury, Chronic/drug therapy , Resveratrol/administration & dosage , Acetaminophen/toxicity , Acute Disease , NF-kappa B/antagonists & inhibitors , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Rats, Sprague-Dawley , Nitric Oxide Synthase/antagonists & inhibitors , Protective Agents , Drug Therapy, Combination , Drug OverdoseABSTRACT
Abstract Objective: Given the high proliferative activity of germinal matrix and its direct correlation with hypoxemia, it is necessary to investigate the possible molecular regulation pathways, to understand the existing clinical relationship between the hypoxic-ischemic insult and the biomarkers NF-kB, AKT-3, Parkin, TRK-C and VEGFR-1. Methods: A hundred and eighteen germinal matrix samples of the central nervous system of patients who died in the first 28 days of life were submitted to histological and immunohistochemistry analysis to identify the tissue immunoexpression of those biomarkers related to asphyxia, prematurity, and death events within 24h. Results: A significantly increased tissue immunoexpression of NF-kB, AKT-3 and Parkin was observed in the germinal matrix of preterm infants. In addition, significantly decreased tissue immunoexpression of VEGFR-1 and NF-kB was observed in patients who experienced asphyxia followed by death within 24 hours. Conclusions: The results suggest a direct involvement between the hypoxic-ischemic insult and NF-kB and VEGFR-1 markers since a decreased immunoexpression of these biomarkers was observed in asphyxiated patients. Furthermore, it is suggested that there was not enough time for VEGFR-1 to be transcribed, translated and expressed on the surface of the plasma membrane. This temporality can be observed in the relationship between NF-kB expression and the survival time of individuals who died within 24 hours, suggesting that this factor is essential for the production of VEGFR-1 and, therefore, to carry out the necessary remodeling effect to neovascularize the affected region.
RESUMO Objetivo: Dada a alta atividade proliferativa da matriz germinativa e sua correlação direta com a hipoxemia, é necessário investigar as possíveis vias de regulação molecular para entender a relação clínica existente entre o insulto hipóxico-isquêmico e os biomarcadores NF-kB, AKT -3, Parkina, TRK-C e VEGFR-1. Métodos: Cento e dezoito amostras de matriz germinativa do sistema nervoso central de pacientes que faleceram nos primeiros 28 dias de vida foram submetidas a análise histológica e imuno-histoquímica para identificar a imunoexpressão tecidual desses biomarcadores relacionados a eventos de asfixia, prematuridade e óbito em 24 horas. Resultados: Observou-se uma imunoexpressão tecidual significativamente aumentada de NF-kB, AKT-3 e Parkin na matriz germinativa de prematuros. Além disso, constatou-se uma imunoexpressão tecidual significativamente diminuída de VEGFR-1 e de NF-kB em pacientes que apresentaram asfixia seguida de morte em 24 horas. Conclusões: Os resultados sugerem o envolvimento direto entre o insulto hipóxico-isquêmico e os marcadores NF-kB e VEGFR-1, visto que se observou uma imunoexpressão diminuída destes biomarcadores nos pacientes asfixiados. Além disso, sugere-se que não houve tempo suficiente para que o VEGFR-1 fosse transcrito, traduzido e expresso na superfície da membrana plasmática. Essa temporalidade pode ser observada na relação entre a expressão de NF-kB e o tempo de vida dos indivíduos que morreram em 24 horas, o que sugere que esse fator é essencial para a produção do VEGFR-1 e, portanto, para realizar o efeito remodelador necessário para neovascularizar a região afetada.
ABSTRACT
SUMMARY: Diabetes is a metabolic disorder characterized by high blood sugar levels and it causes complications in many systems, including the reproductive system. As a result of diabetic conditions, one of the mechanisms that can cause repression of reproductive activity is testicular oxidant stress. The identification of diabetes on the cell signaling molecules axis is still under discussion. The aim of this study was to determine the effect of Transforming Growth Factor (TGFβ), Nuclear Factor kappa B (NF-kB), Heat-schock 90β (HSP90β) signal pathways and E-cadherin cell adhesion molecule on infertility in diabetic rat testicular tissue. In our study, includes histological, molecular and biochemical analysis of testicular tissue removed at the end of the 2 weeks experiment period. A total of 14 adult male rats were divided as control and diabetes. No intervention was given to 7 male rats in the control group. For the diabetic group, 7 male rats were injected by intraperitoneal with a single dose of 55 mg/kg streptozotocin (STZ). TGFβ, NF-kB, HSP90β and E-cadherin proteins were immunohistochemically studied to investigate possible tissue damage, inflammatory process, cell stabilization and integrity due to diabetes. In order to determine oxidant stress, lipid peroxidation product malondialdehyde (MDA), glutathione (GSH) and glutathione peroxidase (GPx) analyzes were performed. Fibrosis, inflammatory changes and loss of spermatogenetic series are prominent findings in the diabetic group. On analysis of all the samples with immunostaining, in the diabetic group, TGFβ and NF-kB immunoexpression significantly increased, while Hsp90β and E-cadherin immunoexpression significantly decreased compared with control groups. Experimental diabetes was found to cause fibrosis, inflammation, disrupting cell adhesion and stabilization in testicular tissue. These results suggest that cellular therapy studies are needed for possible damage.
RESUMEN: La diabetes es una enfermedad metabólica caracterizada por niveles altos de azúcar en sangre y causa complicaciones en muchos sistemas, incluido el sistema reproductivo. Como resultado de las condiciones diabéticas, uno de los mecanismos que puede causar alteraciones en la actividad reproductiva es el estrés oxidativo testicular. La identificación de la diabetes en el eje de las moléculas de señalización celular aún está en discusión. El objetivo de este estudio fue determinar el efecto del factor de crecimiento transformante (TGFβ), el factor nuclear kappa B (NF-kB), las vías de señalización de Heat-Schock 90b (HSP90β) y la molécula de adhesión celular de E-cadherina sobre la infertilidad en testículo de rata diabética. Al término de dos semanas se realizaron análisis histológico, molecular y bioquímico del tejido testicular extraído. Las 7 ratas macho del grupo control no fueron intervenidas. Para el grupo de diabéticos, 7 ratas macho fueron inyectadas por vía intraperitoneal con una dosis única de 55 mg / kg de estreptozotocina (STZ). Se estudiaron inmunohistoquímicamente las proteínas TGFβ, NF-kB, HSP90β y E-cadherina para investigar el posible daño tisular, el proceso inflamatorio, la estabilización celular y la integridad debido a la diabetes. Para determinar el estrés oxidativo, se realizaron análisis del producto de peroxidación lipídica malondialdehído (MDA), glutatión (GSH) y glutatión peroxidasa (GPx). La fibrosis, los cambios inflamatorios y la pérdida de series espermatogenéticas son hallazgos destacados en el grupo de ratas diabéticas. En el análisis de todas las muestras con inmunotinción, en el grupo diabético, la inmunoexpresión de TGFβ y NF-kB aumentó significativamente, mientras que la inmunoexpresión de Hsp90β y e-cadherina disminuyó significativamente en comparación con los grupos control. Se encontró que la diabetes experimental causa fibrosis, inflamación, alteración de la adhesión celular y estabilización en el tejido testicular. Estos resultados sugieren que son necesarios estudios de terapia celular para verificar posibles daños.
Subject(s)
Animals , Male , Rats , Testis/pathology , Diabetes Mellitus, Experimental/metabolism , Testis/metabolism , Immunohistochemistry , Transforming Growth Factors/metabolism , Cadherins/metabolism , NF-kappa B/metabolism , HSP90 Heat-Shock Proteins/metabolismABSTRACT
Menopause complications such as cardiovascular and bone diseases represent a major public health concern. We sought to determine whether a high-fat diet (HFD) can augment ovariectomy-induced bone resorption in a rat model of menopause possibly via the upregulation of the inflammatory biomarkers and dyslipidemia. Rats were either ovariectomized and fed a standard laboratory chow (model group) or were ovariectomized and fed with a HFD for 15 weeks before being sacrificed. Ovariectomy significantly (p<0.05) increased body weight, dyslipidemia, insulin resistance, pro-inflammatory cytokines tumor necrosis factor-a (TNF-α) and interleukin-6 (IL-6), and biomarker of bone resorption, nuclear factor-kB (NF-kB), which were augmented by feeding animals with a HFD. This was confirmed through immunohistochemical study, where ovariectomy induced expression of p65/NF-kB protein in tibia bone sections of the model group, which were augmented by HFD. HFD augments ovariectomy-induced bone resorption through increased inflammatory biomarkers and NF-kB in rats.
Las complicaciones de la menopausia, como las enfermedades cardiovasculares y óseas, representan un importante problema de salud pública. Intentamos determinar si una dieta alta en grasas (HFD) puede aumentar la resorción ósea inducida por ovariectomía en un modelo de menopausia en ratas, a través de la regulación positiva de los biomarcadores inflamatorios y la dislipidemia. Las ratas fueron ovariectomizadas y alimentadas con una comida estándar de laboratorio (grupo modelo) o fueron ovariectomizadas y alimentadas con un HFD durante 15 semanas antes de ser sacrificadas. La ovariectomía aumentó significativamente (p <0,05) el peso corporal, dislipidemia, resistencia a la insulina, citocinas proinflamatorias, factor de necrosis tumoral a (TNF-α) e interleucina-6 (IL-6), y el biomarcador de resorción ósea, factor nuclear-kB (NF-kB), que se aumentaron alimentando animales con un HFD. Esto se confirmó a través del estudio inmunohistoquímico, donde la ovariectomía indujo la expresión de la proteína p65 / NF-kB en secciones de hueso de tibia del grupo modelo, que fueron aumentadas por HFD. HFD aumenta la resorción ósea inducida por ovariectomía a través del aumento de biomarcadores inflamatorios y NF-kB en ratas.
Subject(s)
Animals , Female , Rats , Bone Resorption/pathology , Diet, High-Fat/adverse effects , Triglycerides/analysis , Bone Resorption/etiology , Insulin Resistance , Menopause , Ovariectomy/adverse effects , Rats, Wistar , Disease Models, Animal , Dyslipidemias/complicationsABSTRACT
BACKGROUND: The relationship between nuclear factor (NF)-kB signaling pathway and intervertebral disc degeneration is a hotspot in the field of orthopedics. In-depth investigation on various signaling pathways in the intervertebral disc contributes to understanding the mechanism of intervertebral disc degeneration. OBJECTIVE: To investigate the regulation of the serum containing Yishen Huoxue Tongluo Recipe on the NF-kB signal pathway of human intervertebral disc nucleus pulposus cells under different hydrostatic pressures, attempting to explore the possible therapeutic mechanism and target of Yishen Huoxue Tongluo Recipe in the treatment of degenerative diseases of the intervertebral disc from the perspective of molecular biology. METHODS: Passage 3 Human intervertebral disc nucleus pulposus cells were divided into eight groups and were cultured in the drug-containing serum of Yishen Huoxue Tongluo Recipe. The cells were intervened for 2, 4, and 6 hours under different hydrostatic loading conditions (0.3,1, and 3 MPa). The morphology and growth of nucleus pulposus cells were observed by an inverted phase contrast microscope. Ultrastructural changes of intervertebral disc nucleus pulposus cells were observed by a transmission electron microscopy. Cell counting kit-8 method was used to detect the proliferation activity of nucleus pulposus cells. Annexin V-FITC/Propidium Iodide apoptotic kit was used to double-stain nucleus pulposus cells to detect the cell apoptotic rate. Western blot method was used to detect the changes of NF-kB p65, Collagen II, ADAMTS-4, MMP-13 and Caspase-3 in nucleus pulposus cells. RESULTS AND CONCLUSION: (1) Under the same pressure and time, the morphology and growth of nucleus pulposus cells in the pressure+drug-containing serum groups were better than those in the normal pressure group and the simple pressure groups. Among them, the 0.3 and 1 MPa pressure+drug-containing serum groups had more intact cell morphology and better cell growth than the 3 MPa pressure+ drug-containing serum group. (2) The proliferative activity of nucleus pulposus cells was higher in the pressure+drug-containing serum group, and there was a significant difference between the 0.3 MPa pressure+drug-containing serum group and the 0.3 MPa simple pressure group (P < 0.05). (3) The apoptotic rate of nucleus pulposus cells in the pressure+drug-containing serum group was lower than that in the normal pressure group and simple pressure intervention group (P < 0.05). (4) The expression of Collagen II and Caspase-3 increased in the pressure+drug-containing serum group, while the expression of NF-kappa B p65, ADAMTS-4 and MMP-13 decreased in the pressure+drug-containing serum group (P < 0.05). To conclude, Yishen Huoxue Tongluo Recipe can increase cell activity, reduce cell apoptosis and effectively delay the degeneration of nucleus pulposus cells. Its mechanism is likely to promote the expression of Collagen II and Caspase-3 through the NF-kB signaling pathway of nucleus pulposus cells of intervertebral disc, and inhibit the expression of NF-kB p65, ADAMTS-4 and MMP-13.
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Isatis indigotica Fort. (Ban-Lan-Gen) is an herbal medicine prescribed for influenza treatment. However, its active components and mode of action remain mostly unknown. In the present study, erucic acid was isolated from Isatis indigotica Fort., and subsequently its underlying mechanism against influenza A virus (IAV) infection was investigated in vitro and in vivo. Our results demonstrated that erucic acid exhibited broad-spectrum antiviral activity against IAV resulting from reduction of viral polymerase transcription activity. Erucic acid was found to exert inhibitory effects on IAV or viral (v) RNA-induced pro-inflam-matory mediators as well as interferons (IFNs). The molecular mechanism by which erucic acid with antiviral and anti-inflammatory properties was attributed to inactivation of NF-kB and p38 MAPK signaling. Furthermore, the NF-kB and p38 MAPK inhibitory effect of erucic acid led to diminishing the transcriptional activity of interferon-stimulated gene factor 3 (ISGF-3), and thereby reducing IAV-triggered pro-inflammatory response amplification in IFN-β-sensitized cells. Additionally, IAV- or vRNA-triggered apoptosis of alveolar epithelial A549 cells was prevented by erucic acid. In vivo, erucic acid administration consistently displayed decreased lung viral load and viral antigens expression. Meanwhile, erucic acid markedly reduced CD8+cytotoxic T lymphocyte (CTL) recruitment, pro-apoptotic signaling, hyperactivity of multiple signaling pathways, and exacerbated immune inflammation in the lung, which resulted in decreased lung injury and mortality in mice with a mouse-adapted A/FM/1/47-MA(H1N1) strain infection. Our findings provided a mechanistic basis for the action of erucic acid against IAV-mediated inflammation and injury, suggesting that erucic acid may have a therapeutic potential in the treatment of influenza.
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Objective@#To investigate the regulatory effect of the transcription factor NF-kB1 on the expression of miR-195 in prostate cancer (PCa).@*METHODS@#We analyzed the possibility of NF-kB1 binding to the miR-195 promoter and the expression of NF-kB1 in PCa using the JASPAR and Oncomine databases, respectively, and determined the expressions of NF-kB1 and miR-195 in PCa cells by real-time quantitative PCR after inhibiting the former by interfering RNA targeting NF-kB1. We detected the activity of the luciferase reporter gene after constructing its gene plasmid in the miR-195 promoter region and having it co-transfected with the NF-kB1 plasmid. Then we analyzed the correlation between the expressions of miR-195 and NF-kB1 in the prostate tissue.@*RESULTS@#NF-kB1 was overexpressed in PCa. After inhibition of the expression of NF-kB1, that of miR-195 was increased in PC-3 and DU-145 cell lines, with a negative correlation between the NF-kB1 and miR-195 expressions in the PCa tissue. The results of luciferase reporter gene assay showed direct binding of NF-kB1 to the miR-195 promoter zone.@*CONCLUSIONS@#NF-kB1 regulates the expression of miR-195 in prostate cancer.
Subject(s)
Humans , Male , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , NF-kappa B p50 Subunit/metabolism , Promoter Regions, Genetic , Prostatic Neoplasms/genetics , Transcription Factors/metabolismABSTRACT
Adverse changes occur gradually in the skeletal muscles with age via continuous exposure to oxidative stress. Quercetin, a member of the flavonoids family, possesses anti-oxidative and radical-scavenging activities. Therefore, this study investigated the role of quercetin to modulate age-induced changes in the transcript levels of some apoptosis-related genes in rat's gastrocnemius muscles, up to 15 months-old. Half of the rats at each age (1, 5, 10 and 15 months old) were given a vehicle and the other half was given 200 mg/kg quercetin for 2 weeks, respectively. With the increase of age, vehicle groups showed hyalinization of the muscle fibers and a decrease of the catalase and an increase of the malondialdehyde levels. Down-regulation of Bcl2 gene and up-regulation of both NF-κB and Bax genes were recorded. Interestingly, quercetin groups showed focal hyalinization of the muscle fibers at both 10th and 15th months old. An increase in the catalase and a decrease in malondialdehyde levels, up-regulation of Bcl2 gene and down-regulation of both NF-κB and Bax genes were recorded. In conclusion, quercetin minimized age-induced alteration in the morphological structure and the expression of the apoptosis-related genes via increasing the antioxidant defense in the gastrocnemius muscle.
Subject(s)
Animals , Male , Rats , Role , Apoptosis , Muscle, Skeletal , Muscles , Quercetin/analysis , Flavonoids/analysis , Down-Regulation , Up-Regulation , Oxidative Stress , Antioxidants/adverse effectsABSTRACT
Aim To explore the protective mechanisms of Chaiqinchengqi decoction (CQCQD) on liver injury during severe acute pancreatitis ( SAP) based on TLR4/NF-kB p65 pathway. Methods Thirty-six KM mice were randomly divided into control group (Control) , SAP and SAP + CQCQD (treatment group) (n = 12). The mice in SAP group were injected with 20% L-arginine intraperitoneal
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Aim To investigate the effect of FTY-720 on bleomycin-induced pulmonary fibrosis in mice and its mechanism. Methods Forty specific pathogen-free healthy male BALB/c mice were randomly divided into control group, BLM group, NS + FTY-720 control group, and BLM + FTY-720 group, with 5 mice in each group, and the mice were sacrificed on 7th day and 14th day for a total of 8 groups. Lung tissue inflammation and pulmonary fibrosis were observed by HE staining and Masson staining; Bronchoalveolar lavage fluid (BALF) cells were counted by Swiss-Giemsa staining; BALF protein content was determined by BCA assay; the expression levels of inflammatory cytokines IL-IP and TNF-a in BALF supernatant were measured by ELISA; the expression levels of COL1A1 in lung tissues were detected by immunohistochemis-try; and the expression levels of TGF-|31, p-p38 MAPK and NF-kB were detected by Western blot. Results FTY-720 significantly reduced BLM-induced increase of extracellular collagen deposition in lung tissues of mice, reduced IL-1(3 and TNF-a content in BALF of mice, and reduced the protein content and cell number in BALF of mice; FTY-720 inhibited TGF-(31 activity and p38 MAPK phosphorylation, and then inhibited the expression and activation of NF-kB. Conclusions FTY-720 inhibits bleomycin-induced lung fibrosis in mice by inhibiting the TGF-(31/p38 MAPK/NF-kB signaling pathway.
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Aim To investigate whether Sal B alleviates hypoxic-induced cardiomyocyte injury by regulating the priming of NLRP3 inflammasomes. Methods The effects of Sal B on growth of H9C2 cells were examined by CCK8 assay,then the appropriate concen tration of Sal B was selected. The expression level of LDH was detected by Microliter assay. The expression levels of cTn/IL-lp were measured by Elisa assay. The protein and mRNA levels of TLR4/Myd88/I-RAK1/NF-kB/NLRP3 were detected by Western blot and qPCR. Results Hypoxia intervention notably reduced the viability of H9C2 cells and increased the expression levels of cTn/IL-IP. Besides,the protein and mRNA expression levels of TLR4/Myd88/IRAK1/NF-kB/NLRP3 were significant uP-regulated after hypoxia intervention. However, the viability of H9C2 cells increased, the secretion levels of LDH/cTn/IL-1 p were reduced,and the protein and mRNA levels of TLR4/Myd88/IRAK1/NF-KB/NLRP3 were significant inhibited after pretreated with Sal B. Conclusion Sal B attenuates cardiomyocyte injury by regulating the priming of NLRP3 inflammasome.I.
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Swiprosin-1 (also known as EFhd2) is one of the earliest adaptor proteins found in T lymphocytes. Subsequent studies have found that it is also expressed on macrophages, B lymphocytes, mast cells and other immune cells, and affects the activation, migration, apoptosis and inflammatory response of immune cells, and participates in many signaling pathways 9uch as NF-kB, JAK-STAT, MAPK and calcium signaling. This arti cle briefly reviews the role and mechanism of EFhd2 reported in the regulation of immune inflammatory response in recent years, and provides a theoretical basis for further clarifying EFhd2 as a potential target for new immune regulation.
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GPCBAR1, also known as TGR5 receptor, is one member of the GPCRs family. TGR5, a pattern recognition receptor located on cell membrane, is widely expressed in human beings and animals, playing numerous significant roles in anti-inflammation and immune regulation, energy metabolism, glucose metabolism and anti-cancer. Many signaling pathways are induced by TGR5 upon activated. AKT, NF-kB, ERK, STAT3, cAMP signaling pathways, methods and results of TGR5-mediated signaling pathways and t(ie development of new drugs via TGR5-mediated signaling pathways were reviewed in this paper.
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As our age advances, many changes are seen in our body, such as cellular changes, cardiovascular problems, cerebrovascular diseases, including cancer mediated by inflammation and their mediators such as free radicals (ROS, RNS), cytokines, transcription factors (NF-KB, STAT3) due to altered dietary patterns and digestive disorders. The disease pattern can be suppressed by including anti-inflammatory dietary supplements in our diet to prevent various diseases in geriatric peoples associated with inflammation. Chronic activation of the inflammatory response, defined as inflammation, is the key physio-pathological substrate for anabolic resistance, sarcopenia and frailty in older individuals. Nutrients can theoretically modulate this phenomenon. This article briefs about anti-inflammatory dietary supplements in prevention of diseases associated with inflammation in geriatric people.
ABSTRACT
Obesity is a chronic inflammatory disease that affects millions of people worldwide. Most studies observe the effects of a high-fat diet (HFD) in 10-12 weeks. This work investigated the effects induced by a HFD administered for 6 weeks on the nutritional status of mice and some aspects of the inflammatory response in mouse peritoneal macrophages. Male Swiss Webster mice, 2-3 months of age, were fed a control diet or HFD for 6 weeks. After this period, the mice were euthanized, and peritoneal macrophages were collected for immunoassays and assessment of biochemical parameters. A HFD was associated with increased cholesterol, insulin resistance, C-reactive protein (CRP), leptin, and serum resistin levels. Lipopolysaccharide (LPS)- stimulated adipocyte cultures of animals subjected to a HFD showed increased production of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-1), and interleukin-6 (IL-6). However, peritoneal macrophages of the HFD group showed no changes in the levels of these cytokines. LPS-stimulated peritoneal macrophages from HFD-treated animals showed a reduction in mRNA expression of TNF-α and IL-6, as well as a decrease in expression of the transcription factor nuclear factor-kappa B (NF-kB). In conclusion, HFD treatment for 6 weeks induces similar signs to metabolic syndrome and decreases the capacity of peritoneal macrophages to develop an appropriate inflammatory response to a bacterial component
Subject(s)
Animals , Male , Mice , Macrophages, Peritoneal/classification , Diet, High-Fat/adverse effects , NF-kappa B/pharmacokinetics , Metabolic SyndromeABSTRACT
OBJECTIVE To investigate the influence of high mobility group box-1 protein (HMGB1) inhibition on the expression of receptors for advanced glycation end products (RAGE)/Toll-like receptor 4 (TLR4) signal passway related proteins, RAGE, TLR4, NF-kB, interieukin-1β (IL-1βand tumor necrosis factor-α(TNF-α) in HT22 cells stimulated with amyloid-β25-35 (Aβ25-35). METHODS HT22 cells were stimulated with Aβ25-350, 2.5, 5, 10, 20 and 40 ymol • L"1 for 24 h, and the vitality of HT22 cells was determined using MTT assay. Half maximal inhibitory concentration (IC50) was calculated. HT22 cells were divided into 5 groups: Normal cell control group, Apas-K 40 Mmol-L"' group, siRNA 50 Mmol-L"' group, siRNA or scramble siRNA+APas-as group (HT22 cells were transfected with siRNA or scramble siRNA at 50 pmol • L"' for 24 h, then treated with synthetic Apa-as at a final concentration of 40 pmol • L"1 for 24 h). The morphology of HT22 cells was observed under a microscope. The location of HMGB1 was observed by immunofluorescence. The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 were detected by Western blotting. The levels of IL-IP and TNF-α in the culture supernatants were examined by ELISA. RESULTS The ICso of HT22 cells treated with Apas-as for 24 h was 41.17 Mmol-L-1, and A25-35 40 Mmol-L"1 was selected in subsequent experiments. After 24 h treatment with A25-35 40 nmol-L"', a large number of cells died, or aggregated into clusters, processes decreased, cell gaps increased, and HMGB1 was released from the nucleus to the outside. The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells were significantly increased (P<0.05) after treatment with Apas-as, and the levels of IL-1p and TNF-α in the culture supernatants were significantly increased (P<0.05). The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells were significantly decreased (P<0.05) after 24 h treatment with siRNA 50 pmol-L"' and the levels of IL-1p and TNF-α in the culture supernatants were significantly decreased (P<0.05). CONCLUSION RNA interference with HMGB1 reduces the expression of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells stimulated with A25-35
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OBJECTIVE To investigate the effects of bilobalide (BB) on the myocardial tissue of rats with myocardial ischemia/reperfusion (Ml/R). METHODS An Ml/R model was prepared by ligating the anterior descending branch of the left coronary artery in rats for 30 min. BB 2, 4 and 8 mg-kg"1 was adminsterted for four weeks. The heart rate of rats was recorded and myocardial infarction area was detected. Detective kits were used to evaluate the concentrations of creatine kinase MB isoenzyme (CK-MB) and lactate dehydrogenase (LDH). HE staining was performed to observe the myocardial injury, and TUNEL staining was performed to evaluate apoptosis. ELISA was performed to determine the concentrations of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), IL-1p, tumor necrosis factor-a (TNF-a) and IL-10. Immunohistochemistry was performed to evaluate the expressions of Ki67 and interleukin-6 (IL-6). Western blotting was performed to evaluate the expressions of Bcl-2, Bax, cleaved-caspase 3, nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2), heme oxygenase 1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQ01), NF-kB P65, phosphorylated inhibitor of NF-kB (P-IkBoc) and IkBcx kinase (p-IKK). RESULTS Compared with normal control group, the percentage of the myocardial infarction area and the concentration of CK-MB and LDH were increased in Ml/R model group, but heart rate was decreased (P<0.01) and myocardial tissue showed myocardial fiber rupture and inflammatory infiltration. Compared with Ml/R model group, the percentage of the myocardial infarction area and the concentration of CK-MB and LDH were decreased in Ml/R model+BB 2, 4 and 8 mg-kg"1 groups (P<0.05), but heart rate was increased (P<0.01) and myocardial tissue fibers tended to be regular while inflammatory cell infiltration was significantly reduced. Compared with normal control group, the percentage of apoptotic cells and Ki67 expression-positive cells and the expressions of Bax and cleaved-caspase 3 were increased in Ml/R model group (F<0.01), but the expression of Bcl-2 was decreased markedly (P<0.01). Compared with Ml/R model group, the percentages of apoptotic cells and Ki67 expression-positive cells and the expressions of Bax and cleaved-caspase 3 were decreased in Ml/R model+BB 2, 4 and 8 mg∗kg-1 groups (P<0.01), while the expression of Bcl-2 was increased (P<0.01). Ml/R could lower the activity of SOD and GSH-Px, but enhance MDA content. BB could weaken the effect of Ml/R on SOD, GSH-Px and MDA. Compared with normal control group, the concentrations of IL-1β, TNF-α and IL-10 in serum and the percentage of IL-6 expression-positive cells in myocardial tissue of Ml/R model group were increased (P<0.01). Compared with Ml/R model group, the concentrations of IL-1β and TNF-a and the percentage of IL-6 expression-positive cells were decreased in Ml/R model∗ BB 2, 4 and 8 mg-kg"1 group (P<0.01), but the concentration of IL-10 was increased(P<0.05). Compared with mormal control group, the protein levels of Nrf2, HO-1 and NQ01 significantly decreased (P<0.01), while the levels of NF-kB P65, p-IKK and p-kBa significantly increased (P<0.01). BB could effectively reverse protein changes (P<0.05, P<0.01). CONCLUSION BB can protect rats with Ml/R via anti-inflamma-tion and anti-oxidation.
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OBJECTIVE To explore the inhibition of Compound Kushen Injection (CKI) on the prolif-eration, in vitro migration and in vitro invasion of SMMC-7221 cells and the mechanism. METHODS SMMC-7221 cells were treated with CKI of different dilution percentages (0.0% (cell control), 2.5%, 5.0% and 10.0%) for 24 h. The viability of SMMC-7221 cells was evaluated by MTT assay, while the ability of cell migration and invasion was evaluated by the wound healing and transwell chamber assay, respectively. Cells were treated with 0.0%, 5.0% and 10.0% CKI for 12 or 24 h, before the mRNA and protein levels of NF-kB were detected by RT-PCR and Western blotting, respectively. RESULTS Compared with the cell control group, the survival rates in 2.5%, 5.0% and 10.0% CKI groups were reduced to (92±7)%, (77±5)% (P<0.01) and (56±7)% (P<0.01), the migration rates to (90±7)%, (50±10)% (P<0.01) and (25±5)% (P<0.01), and the invasion rates to (98±7)%, (51 ±10)% (P<0.01) and (20±5)% (P<0.01), respectively. The NF-kB mRNA levels in 5.0% and 10.0% CKI groups were reduced to (42±9)% and (46±10)% of the cell control group (P<0.01) after 12 h treatment of CKI, and the protein levels of NF-kB were reduced to (67±16)% and (27±11)% of cell control group after 24 h treatment with CKI, respectively (P<0.01). CONCLUSION CKI can inhibit the proliferation, migration and invasion of SMMC-7221 cells in vitro, and the mechanism may be related to the inhibition of NF-kB signal pathway.
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Aim To study the intervention effect of tet- ramethylpyrazine(TMP) on human microvascular endothelial cells (HMECs) inflammatory response induced by activated complement alternative pathway and the possible molecular mechanisms. Methods HMECs were pretreated with different concentrations of TMP, and then exposed to the activated products of the complement alternative pathway which was prepared by cobra venom factor( CVF). The supernatant was removed and assayed for expression of the adhesion molecules (ICAM-1, VCAM-1 and E-selectin) and the inflammatory mediator(IL-6 and TNF-ot) by using ELISA reagent kits. The nucleus transcriptional activity of NF- kB was measured by the dual luciferase reporter assay ? system. Results The adhesion molecules, inflammato ry mediator and nucleus transcriptional activity of NF- kB increased after HMECs were exposed to the products of the activated complement alternative pathway. The up-regulation of ICAM-1, VCAM-1, E-selectin, IL-6, TNF-a and the nucleus transcriptional activity of NF-kB were inhibited by various concentrations of TMP in a dose-dependent manner. Conclusions TMP can effectively reduce inflammatory response of HMECs in-duced by the activated complement alternative pathway , and the mechanism may be highly related to inhibition of nucleus transcriptional activity of NF-kB.