Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Association of serum irisin levels with endothelial dysfunction markers in an in vitro model of HUVEC cells challenged with serum from pediatric patients with Diabetes Mellitus and Metabolic Syndrome(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-05-12) Luna Ceron, Eder; SEGURA MEDINA, PATRICIA; 15820; Segura Medina, Patricia; Elizondo Montemayor, Leticia; emipsanchez; Sobrevia, Luis; García Rivas, Gerardo de Jesus; Guerrero Beltrán, Carlos Enrique; School of Medicine and Health Sciences; Campus Monterrey; Gonzalez Castillo, Elena CristinaBackground and aims: Endothelial dysfunction is a crucial pathophysiological mechanism for cardiovascular diseases and a potential consequence of metabolic disorders. Preclinical studies have revealed that irisin, an adipomyokine, can exert beneficial effects in preventing endothelial dysfunction by decreasing the production of oxidative stress, promoting endothelial production of nitric oxide (NO), and increasing the endothelial cell viability. However, these studies have the limitation of studying single-damage inductors for endothelium, evading the representativeness of the combined adverse metabolic extracellular milieu in humans. We first aimed to characterize the role of irisin on dual damage in an in vitro model of hyperglycemia and hyperlipidemia. We further investigated the associations between irisin serum levels and endothelial dysfunction markers in an in vitro model of human umbilical vein endothelial cells (HUVEC) challenged with the serum from pediatric patients diagnosed with Type 2 Diabetes Mellitus (T2DM) and Metabolic Syndrome (MetS). Methodology: We characterized the effects of recombinant irisin (0-20 nM) after 24 hours of treatment with high glucose and high palmitic acid on cell viability, cell migration and adhesion, and endothelial NO production. In a second stage, we obtained serum samples from a cohort of 55 pediatric patients aged 6 to 16 years old. These patients were grouped depending on their metabolic status into T2DM patients (n=21), MetS (n=18) patients, and healthy controls (n=16). We performed cell viability assays and measured endothelial NO production in individual HUVEC cell cultures challenged for 24 hours with the serum of these patients. Results: Treatment with recombinant irisin (10 nM) demonstrated a significant maintenance of endothelial cell viability in culture with hyperlipidemic and hyperglycemic media. Endothelial cell viability was supported mainly by significantly reducing late-apoptotic cell populations. Functionally, irisin remarkably maintained the endothelial NO production in these conditions while improving endothelial cell adhesion to a 5% gelatin extracellular matrix substrate. Assays involving patients revealed significantly lower serum irisin levels in the MetS and T2DM groups than in controls. HUVEC cell cultures challenged with the serum of patients with T2DM and MetS showed substantially lower endothelial NO production, lower cell viability, and higher rates of late apoptotic and necrotic cells. We observed significant positive associations between irisin serum levels and NO production (R= 0.27, p<0.05) and cell viability (R=0.37, p<0.05). Negative correlations between serum irisin levels and rates of late apoptotic cells (R=-0.41) and necrotic cells (R=-0.32, p<0.05) were identified. Conclusions: Overall, these results suggest that irisin exerts beneficial effects on endothelial integrity in the presence of adverse metabolic scenarios by maintaining endothelial cell functionality through nitric oxide production, sustaining cell viability, downregulating their late apoptosis. This is the first study to show that serum irisin levels are positively correlated with endothelial cell viability and nitric oxide production, while inversely correlated with late-apoptotic and necrotic endothelial cell rate. The loss of these protection mechanisms in patients with MetS and T2DM, which show low circulant irisin levels, translates into the detrimental functional effects on endothelial cells and their association with endothelial dysfunction markers.
- The role of irisin on the polarization of monocyte-derived macrophages from pediatric patients with type 2 diabetes mellitus(Instituto Tecnológico y de Estudios Superiores de Monterrey) González Gil, Adrián Marcelo; Elizondo Montemayor, Luz Leticia; puelquio; García Rivas, Gerardo de Jesús; Estrada, Manuel; Genevieve Brunck, Marion Emilie; School of Medicine and Health Sciences; Campus Monterrey; González Castillo, Elena CristinaIrisin is a novel peptide hormone released from skeletal muscle following acute bouts of physical exercise. Besides its originally described role on adipose tissue browning, its direct antiinflammatory properties have been recently described. Specifically, when murine RAW264.7 macrophages were pre-treated with irisin, both gene and protein expression of key proinflammatory cytokines, such as TNF-α, IL-1β, and IL-6, decrease following exposure to lipopolysaccharide (LPS), while levels of the anti-inflammatory cytokine IL-10 increase. These changes have been linked to decreased TLR4 protein expression and activity of NF-ΚB. As well, in murine models, irisin has been described to favor an alternatively activated M2 macrophage phenotype, both from a M0 unstimulated state and a M1 classical proinflammatory state. Recently, the importance of an anti-inflammatory microenvironment to maintain the differentiation and function of brown adipose tissue has been emphasized in preclinical models, which could imply that irisin exerts its global metabolic effects indirectly through its action on adipose tissue macrophages, at least partially. Thus, irisin and its downstream pathways could represent a novel therapeutic target in type 2 diabetes mellitus (T2DM), a disease state characterized by a systemic low-grade proinflammatory state and lower circulating levels of irisin compared with healthy controls. In humans, evidence shows that M1 macrophage infiltrates in adipose tissue originate from circulating monocytes, but no studies have described the role of irisin on any type of human immune cell. Using monocyte-derived macrophages (MDMs) obtained from peripheral blood mononuclear cells (PBMCs) from a previously characterized cross-sample of pediatric T2DM patients (n=15) and matched healthy controls (n=8), we sought to determine differences in MDM immunophenotypes between groups, their response to in vitro treatment with recombinant human irisin, and to correlate patients’ basal monocyte subsets with their clinical and biochemical parameters and MDM phenotypes. In order to standardize the procedures previous to testing cryopreserved samples of T2DM and control pediatric patients, blood was obtained from one healthy adult individual. In this subject, in vitro irisin treatment significantly increased CD163 MFI. T2DM subjects had higher proportions of circulating intermediate monocytes (IMs) relative to healthy controls, which correlated positively with body fat percentage and the inflammatory marker hs-CRP and negatively with HDL-c. MDMs from T2DM subjects had similar polarization profiles compared with controls when exposed to IL-4 and IFN-γ and LPS. However, macrophage polarization capacity, as measured by M1 (CD80) and M2 (CD163, CD200R) marker MFI, was significantly associated with basal monocyte proportions when considering all participants. Upon irisin treatment, CD163 upregulation was no longer observed in MDMs from patient samples, but a trend towards decreased NF-κB activation was noted. Our results provide preliminary evidence in favor of irisin’s anti-inflammatory role in human macrophages but must be replicated in future studies with larger sample sizes. On the other hand, increased IMs in pediatric T2DM might suggest enhanced monocyte migration and differentiation to macrophages in obese white adipose tissue or to vascular atherosclerotic lesions early in disease evolution, which warrants future longitudinal and mechanistic studies.