RESUMO
MicroRNAs (miRNA) have emerged as important post-transcriptional regulators of metabolic pathways that contribute to cellular and systemic lipoprotein homeostasis. Here, we identify two conserved miRNAs, miR-224, and miR-520d, which target gene networks regulating hepatic expression of the low-density lipoprotein (LDL) receptor (LDLR) and LDL clearance. In silico prediction of miR-224 and miR-520d target gene networks showed that they each repress multiple genes impacting the expression of the LDLR, including the chaperone molecules PCSK9 and IDOL that limit LDLR expression at the cell surface and the rate-limiting enzyme for cholesterol synthesis HMGCR, which is the target of LDL-lowering statin drugs. Using gain- and loss-of-function studies, we tested the role of miR-224 and miR-520d in the regulation of those predicted targets and their impact on LDLR expression. We show that overexpression of miR-224 or miR-520d dose-dependently reduced the activity of PCSK9, IDOL, and HMGCR 3'-untranslated region (3'-UTR)-luciferase reporter constructs and that this repression was abrogated by mutation of the putative miR-224 or miR-520d response elements in the PCSK9, IDOL, and HMGCR 3'-UTRs. Compared to a control miRNA, overexpression of miR-224 or miR-520d in hepatocytes inhibited PCSK9, IDOL, and HMGCR mRNA and protein levels and decreased PCSK9 secretion. Furthermore, miR-224 and miR-520d repression of PCSK9, IDOL, and HMGCR was associated with an increase in LDLR protein levels and cell surface expression, as well as enhanced LDL binding. Notably, the effects of miR-224 and miR-520d were additive to the effects of statins in upregulating LDLR expression. Finally, we show that overexpression of miR-224 in the livers of Ldlr +/- mice using lipid nanoparticle-mediated delivery resulted in a 15% decrease in plasma levels of LDL cholesterol, compared to a control miRNA. Together, these findings identify roles for miR-224 and miR-520d in the posttranscriptional control of LDLR expression and function.
RESUMO
During obesity, macrophages infiltrate the visceral adipose tissue and promote inflammation that contributes to type II diabetes. Evidence suggests that the rewiring of cellular metabolism can regulate macrophage function. However, the metabolic programs that characterize adipose tissue macrophages (ATM) in obesity are poorly defined. Here, we demonstrate that ATM from obese mice exhibit metabolic profiles characterized by elevated glycolysis and oxidative phosphorylation, distinct from ATM from lean mice. Increased activation of HIF-1α in ATM of obese visceral adipose tissue resulted in induction of IL-1ß and genes in the glycolytic pathway. Using a hypoxia-tracer, we show that HIF-1α nuclear translocation occurred both in hypoxic and non-hypoxic ATM suggesting that both hypoxic and pseudohypoxic stimuli activate HIF-1α and its target genes in ATM during diet-induced obesity. Exposure of macrophages to the saturated fatty acid palmitate increased glycolysis and HIF-1α expression, which culminated in IL-1ß induction thereby simulating pseudohypoxia. Using mice with macrophage-specific targeted deletion of HIF-1α, we demonstrate the critical role of HIF-1α-derived from macrophages in regulating ATM accumulation, and local and systemic IL-1ß production, but not in modulating systemic metabolic responses. Collectively, our data identify enhanced glycolysis and HIF-1α activation as drivers of low-grade inflammation in obesity.
Assuntos
Tecido Adiposo Branco/metabolismo , Glicólise , Subunidade alfa do Fator 1 Induzível por Hipóxia/fisiologia , Interleucina-1beta/biossíntese , Gordura Intra-Abdominal/metabolismo , Macrófagos/metabolismo , Obesidade/metabolismo , Tecido Adiposo Branco/patologia , Animais , Medula Óssea/patologia , Hipóxia Celular/genética , Células Cultivadas , Dieta Hiperlipídica/efeitos adversos , Regulação da Expressão Gênica , Glicólise/efeitos dos fármacos , Glicólise/genética , Interleucina-1beta/genética , Gordura Intra-Abdominal/patologia , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Obesidade/genética , Especificidade de Órgãos , Fosforilação Oxidativa , Palmitatos/farmacologiaRESUMO
The purpose of the study is to examine how several well-known correlates of nonsuicidal self-injury (NSSI) might work together to contribute to the occurrence of this behavior. Specifically, we examined models including child abuse, psychiatric comorbidity, and disinhibition, testing how these factors may work together to lead to NSSI in the past month. Participants (n=194; 144 female; age 13-18 years) were recruited from a short-term, acute adolescent residential unit. Within 48 hours of admission to the hospital participants completed structured clinical interviews assessing mental disorders and patterns of NSSI. Following the interviews, participants completed a self-report questionnaire assessing childhood abuse and a computerized continuous performance task. Consistent with study hypotheses, results revealed that the association between child abuse and NSSI is partially mediated by comorbidity. Although disinhibition is associated with comorbidity, contrary to our hypothesis, disinhibition does not mediate the relation between child abuse and NSSI. Collectively, these findings provide new information about how comorbidity may increase risk for NSSI, and critically, discuss the potential importance of creating targeted programs to reduce the prevalence of child abuse.
Assuntos
Comportamento do Adolescente/psicologia , Maus-Tratos Infantis/psicologia , Inibição Psicológica , Comportamento Autodestrutivo/psicologia , Adolescente , Feminino , Humanos , Masculino , Testes Neuropsicológicos , Fatores de Risco , Autorrelato , Inquéritos e QuestionáriosRESUMO
During obesity, macrophage accumulation in adipose tissue propagates the chronic inflammation and insulin resistance associated with type 2 diabetes. The factors, however, that regulate the accrual of macrophages in adipose tissue are not well understood. Here we show that the neuroimmune guidance cue netrin-1 is highly expressed in obese but not lean adipose tissue of humans and mice, where it directs the retention of macrophages. Netrin-1, whose expression is induced in macrophages by the saturated fatty acid palmitate, acts via its receptor Unc5b to block their migration. In a mouse model of diet-induced obesity, we show that adipose tissue macrophages exhibit reduced migratory capacity, which can be restored by blocking netrin-1. Furthermore, hematopoietic deletion of Ntn1 facilitates adipose tissue macrophage emigration, reduces inflammation and improves insulin sensitivity. Collectively, these findings identify netrin-1 as a macrophage retention signal in adipose tissue during obesity that promotes chronic inflammation and insulin resistance.