Regulación de la vía de tnf-a/nfkb por acción de la glicina en adipocitos
Introduction: Glycine, an amino acid non-essential and simple structure consisting of a hydrogen as a side chain (Lynch, 2004); it has shown to be protective against cell injury, providing cytoprotection (Zhong et al., 2003). Likewise, it partnered with the blocking of the systemic inflammatory proc...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2018 |
| País: | México |
| Institución: | Universidad Autónoma Metropolitana |
| Repositorio: | Repositorio Institucional de la UAM Iztapalapa |
| Idioma: | español |
| OAI Identifier: | oai:bindani.izt.uam.mx:g732d898n |
| Acceso en línea: | https://doi.org/10.24275/uami.g732d898n |
| Access Level: | acceso abierto |
| Palabra clave: | info:eu-repo/classification/LEM/Glycine info:eu-repo/classification/LEM/Adipocitos info:eu-repo/classification/LEM/Glicina info:eu-repo/classification/cti/2 |
| Sumario: | Introduction: Glycine, an amino acid non-essential and simple structure consisting of a hydrogen as a side chain (Lynch, 2004); it has shown to be protective against cell injury, providing cytoprotection (Zhong et al., 2003). Likewise, it partnered with the blocking of the systemic inflammatory process that originates in a variety of pathological states. These processes involve powerful inflammatory mediators, such as cytokines, which play an important role in the progressive inflammatory response (Matilla et al., 2002; Zhong et al., 2003). The effects of glycine are regulated by a specific receptor (GlyR), which is anchored to the plasma membrane and associated with Cl- channels. When there is the union of glycine to your receiver, it exerts its inhibitory action (Chatterton et al., 2002; Laube et al., 2002); such action of the glycine can be selectively blocked by strychnine (Sakata et al., 2001). So far investigations on the effects of glycine in vitro and in vivo studies have shown that decreases the expression and secretion of proinflammatory cytokines and increases the anti-inflammatory (Alarcón-Aguilar et al., 2008; Almanza-Perez et al., 2010; Garcia - Macedo et al., 2008). Effects that have been associated with the participation of the transcription factor NF-kB (nuclear factor kappa B), which promotes transcription of genes encoding proinflammatory cytokines and chemokines, which have as main function to recruit macrophages polymorphonuclear site of tissue damage, to kill the invader by phagocytosis or trying to regulate the inflammatory process with the synthesis of anti-inflammatory cytokines (Hayden and Ghosh, 2004). It has been shown that pretreatment with glycine suppresses the activation of this vii transcription factor as well as the activity of its inhibitor and the complex IKK-a resulting in a decrease in the synthesis of proinflammatory cytokines and an improvement in the profile inflammatory (Blancas-Flores et al., 2012; ContrerasNuñez., 2013). This research intends to continue with the elucidation of the mechanism of action of glycine in adipocytes explaining his participation in the inflammatory process. General objective: determine if the anti-inflammatory effect of glycine occurs by inhibition of IKK-a/b complex on via TNF-a/NF-kB by binding to a specific receiver. Material and methods: We used the cell line 3T3-L1; the cells were incubated with TNF - (5 ng/mL) for 30 min (positive control), BAY (10 µM, an inhibitor of NF-kB) during 30 min (negative control), and pretreatment with glycine (10 mM) to 15, 30, 45, 60, 120 and 240 min. He was the extraction of nuclear and cytoplasmic protein for its quantification. Applied technique of the Western Blot for detection of protein kinases (IKK-a/b). The activation of NF-kB as well as the secretion of cytokines (IL-6, TNF-a and adiponectin) was measured by ELISA method. He was the extraction of mRNA and applied the technique of RT-PCR for the quantification of cytokines (IL-6, TNF-a and adiponectin) and the expression of glycine receptor. The [Ca2+]i and determined the location of the receiver of glycine in the membrane by confocal microscopy. In trials to determine a possible presence of receptors for glycine (glycine receptor antagonist) strychnine was used by the ELISA method in which measured the activation of NF-kB. The data were analyzed using an analysis of variance followed by complementary Tukey-Kramer test, with a significance level of 95%. Results: The results showed that glycine decreased the presence of IKK-a at 30, 60 and 120 min, as well as the p-IKK-a (Thr 23) to 15, 60, 120, 240 min. However, the complex IKK-a/b decreased its presence at 15 min and showed a significant increase at 240 min relative to the positive control TNF-a, there was also a decrease in the content of p-IKK-a/b (Ser176/177) at 30, 60, 120 and 240 min. This coincided with a decrease in phosphorylation of p65 NF-kB (pS536). Effect that caused a decrease in the expression and secretion of IL-6 and TNF-a. Expression and secretion of adiponectin were increased. It also showed that glycine generates a decrease in the [Ca+2]i increasing the expression of mRNA of receptor (GlyR) glycine, as well as its location in the Adipocyte plasma membrane. Conclusión: IKK-a/b is regulated by the action of glycine, interfering in their activity and, consequently, by suppressing the activation of NF-kB, action associated with a specific receptor that causes a hyperpolarization in the plasma membrane of the adipocyte, generating blockade of voltage-dependent channels, as of Ca+2, regulating the expression and secretion of proinflammatory cytokines (IL-6, TNF-a) and increasing an anti-inflammatory cytokine (adiponectin) in 3T3-L1 adipocytes, contributing to the reduction of the inflammatory process at the systemic level. |
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