RESUMEN
Impairments in mitochondrial function and substrate metabolism are implicated in the etiology of obesity and Type 2 diabetes. MicroRNAs (miRNAs) can degrade mRNA or repress protein translation and have been implicated in the development of such disorders. We used a contrasting rat model system of selectively bred high- (HCR) or low- (LCR) intrinsic running capacity with established differences in metabolic health to investigate the molecular mechanisms through which miRNAs regulate target proteins mediating mitochondrial function and substrate oxidation processes. Quantification of select miRNAs using the rat miFinder miRNA PCR array revealed differential expression of 15 skeletal muscles (musculus tibialis anterior) miRNAs between HCR and LCR rats (14 with higher expression in LCR; P < 0.05). Ingenuity Pathway Analysis predicted these altered miRNAs to collectively target multiple proteins implicated in mitochondrial dysfunction and energy substrate metabolism. Total protein abundance of citrate synthase (CS; miR-19 target) and voltage-dependent anion channel 1 (miR-7a target) were higher in HCR compared with LCR cohorts (~57 and ~26%, respectively; P < 0.05). A negative correlation was observed for miR-19a-3p and CS (r = 0.32, P = 0.015) protein expression. To determine whether miR-19a-3p can regulate CS in vitro, we performed luciferase reporter and transfection assays in C2C12 myotubes. MiR-19a-3p binding to the CS untranslated region did not change luciferase reporter activity; however, miR-19a-3p transfection decreased CS protein expression (â¼70%; P < 0.05). The differential miRNA expression targeting proteins implicated in mitochondrial dysfunction and energy substrate metabolism may contribute to the molecular basis, mediating the divergent metabolic health profiles of LCR and HCR rats.
Asunto(s)
Tolerancia al Ejercicio/genética , MicroARNs/metabolismo , Mitocondrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Carrera , Animales , Western Blotting , Línea Celular , Citrato (si)-Sintasa/metabolismo , Metabolismo Energético/genética , Técnicas In Vitro , Ratones , Fibras Musculares Esqueléticas/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Canal Aniónico 1 Dependiente del Voltaje/metabolismoRESUMEN
We compared the impact of a high versus low energy intake first meal on glucose and insulin responses during prolonged sitting in individuals with prediabetes. Thirteen adults with overweight/obesity and prediabetes (mean ± SD age: 60 ± 6 years, BMI: 33 ± 4 kg/m²; 2 h OGTT: 8.9 ± 1.1 mmol/L) completed two randomised trials: 10 h uninterrupted sitting, incorporating three meals with matching macronutrient compositions but different energy distributions: High-Energy Breakfast (HE-BF; breakfast: 50%, lunch: 30%, dinner: 20% energy intake), Low-Energy Breakfast (LE-BF: 20%/30%/50% energy intake). Venous blood was sampled from 08:00â»18:00 h for determination of plasma glucose and insulin concentrations, with 24 h continuous glucose monitoring (CGM). Total glucose area under the curve (AUC; +5.7 mmol/L/h, p = 0.019) and mean plasma glucose concentrations (+0.5 mmol/L, p = 0.014) were greater after HE-BF compared to LE-BF. In the HE-BF condition, compared to LE-BF, there was a greater incremental area under the curve (iAUC) for plasma glucose post-breakfast (+44 ± 59%, p = 0.007), but lower iAUC post-lunch (−55 ± 36%, p < 0.001). Total insulin AUC was greater (+480 mIU/mL/h, p < 0.01) after HE-BF compared to LE-BF. Twenty-four-hour (24 h) CGM revealed no differences in mean glucose and total AUC between conditions. Compared to a low-energy first meal, a high-energy first meal elicited exaggerated plasma insulin and glucose responses until lunch but had little effect on 24 h glycaemia. During periods of prolonged sitting, adults with prediabetes may have more beneficial postprandial insulin responses to a low-energy first meal.