We investigated the protective effect of Brazilian propolis, a natural resinous substance produced by honeybees, against glycation stress in mouse skeletal muscles. Mice were divided into four groups: (1) Normal diet + drinking water, (2) Brazilian propolis (0.1%)-containing diet + drinking water, (3) normal diet + methylglyoxal (MGO) (0.1%)-containing drinking water, and (4) Brazilian propolis (0.1%)-containing diet + MGO (0.1%)-containing drinking water. MGO treatment for 20 weeks reduced the weight of the extensor digitorum longus (EDL) muscle and tended to be in the soleus muscle. Ingestion of Brazilian propolis showed no effect on this change in EDL muscles but tended to increase the weight of the soleus muscles regardless of MGO treatment. In EDL muscles, Brazilian propolis ingestion suppressed the accumulation of MGO-derived advanced glycation end products (AGEs) in MGO-treated mice. The activity of glyoxalase 1 was not affected by MGO, but was enhanced by Brazilian propolis in EDL muscles. MGO treatment increased mRNA expression of inflammation-related molecules, interleukin (IL)-1β, IL-6, and toll-like receptor 4 (TLR4). Brazilian propolis ingestion suppressed these increases. MGO and/or propolis exerted no effect on the accumulation of AGEs, glyoxalase 1 activity, and inflammatory responses in soleus muscles. These results suggest that Brazilian propolis exerts a protective effect against glycation stress by inhibiting the accumulation of AGEs, promoting MGO detoxification, and reducing proinflammatory responses in the skeletal muscle. However, these anti-glycation effects does not lead to prevent glycation-induced muscle mass reduction.

5′‐Adenosine monophosphate‐activated protein kinase (AMPK) has been identified as a key mediator of contraction‐stimulated insulin‐independent glucose transport in skeletal muscle. Caffeine acutely stimulates AMPK in resting skeletal muscle, but it is unknown whether caffeine affects AMPK in contracting muscle. Isolated rat epitrochlearis muscle was preincubated and then incubated in the absence or presence of 3 mmol/L caffeine for 30 or 120 min. Electrical stimulation (ES) was used to evoke tetanic contractions during the last 10 min of the incubation period. The combination of caffeine plus contraction had additive effects on AMPKα Thr¹⁷² phosphorylation, α‐isoform‐specific AMPK activity, and 3‐O‐methylglucose (3MG) transport. In contrast, caffeine inhibited basal and contraction‐stimulated Akt Ser⁴⁷³ phosphorylation. Caffeine significantly delayed muscle fatigue during contraction, and the combination of caffeine and contraction additively decreased ATP and phosphocreatine contents. Caffeine did not affect resting tension. Next, rats were given an intraperitoneal injection of caffeine (60 mg/kg body weight) or saline, and the extensor digitorum longus muscle was dissected 15 min later. ES of the sciatic nerve was performed to evoke tetanic contractions for 5 min before dissection. Similar to the findings from isolated muscles incubated in vitro, the combination of caffeine plus contraction in vivo had additive effects on AMPK phosphorylation, AMPK activity, and 3MG transport. Caffeine also inhibited basal and contraction‐stimulated Akt phosphorylation in vivo. These findings suggest that caffeine and contraction synergistically stimulate AMPK activity and insulin‐independent glucose transport, at least in part by decreasing muscle fatigue and thereby promoting energy consumption during contraction.

Exercise has beneficial effects on our health by stimulating metabolic activation of skeletal muscle contraction. Caffeine is a powerful metabolic stimulant in the skeletal muscle that has ergogenic effects, including enhanced muscle power output and endurance capacity. In the present study, we aim to characterize the metabolic signatures of contracting muscles with or without caffeine stimulation using liquid chromatography-mass spectrometry and capillary electrophoresis coupled to mass spectrometry. Isolated rat epitrochlearis muscle was incubated in the presence or absence or of 3 mM caffeine for 30 min. Electrical stimulation (ES) was used to induce tetanic contractions during the final 10 min of incubation. Principal component analysis and hierarchical clustering analysis detected 184 distinct metabolites across three experimental groups—basal, ES, and ES with caffeine (ES + C). Significance Analysis of Microarray identified a total of 50 metabolites with significant changes in expression, and 23 metabolites significantly changed between the ES and ES + C groups. Changes were observed in metabolite levels of various metabolic pathways, including the pentose phosphate, nucleotide synthesis, β-oxidation, tricarboxylic acid cycle, and amino acid metabolism. In particular, D-ribose 5-phosphate, IMP, O-acetylcarnitine, butyrylcarnitine, L-leucine, L-valine, and L-aspartate levels were higher in the ES + C group than in the ES group. These metabolic alterations induced by caffeine suggest that caffeine accelerates contraction-induced metabolic activations, thereby contributing to muscle endurance performance and exercise benefits to our health.

Objective: 5’-adenosine monophosphate-activated protein kinase (AMPK) has been identified as a key mediator of contraction-stimulated insulin-independent glucose transport in skeletal muscle. Caffeine acutely stimulates AMPK in resting skeletal muscle, but it is unknown whether caffeine affects AMPK in contracting muscle. In this study, we examined the effect of caffeine stimulation on contraction-stimulated AMPK activity and glucose transport.Materials and Methods: (1) Isolated rat epitrochlearis muscle was preincubated and then incubated in the absence or presence of 3 mM caffeine for 30 min. Electrical stimulation (ES) was used to evoke tetanic contractions during the last 10 min of the incubation period. (2) Rats were given an intraperitoneal injection of caffeine (60 mg/kg body weight) or saline, and the extensor digitorum longus muscle was dissected 15 min later. ES of the sciatic nerve was performed to evoke tetanic contractions for 5 min before dissection.Results: (1) The combination of caffeine plus contraction had additive effects on AMPKα Thr 172 phosphorylation, α-isoform-specific AMPK activity, and 3-O-methylglucose (3MG) transport. Caffeine significantly delayed muscle fatigue during contraction, and the combination of caffeine and contraction additively decreased ATP and phosphocreatine contents. (2) Similar to the findings from isolated muscles incubated in vitro, the combination of caffeine plus contraction in vivo had additive effects on AMPK phosphorylation, AMPK activity, and 3MG transport.Conclusions: These findings suggest that caffeine and contraction synergistically stimulate AMPK activity and insulin-independent glucose transport, at least in part by decreasing muscle fatigue and thereby promoting energy consumption during contraction.