Physiological Doses of Hydroxytyrosol Modulate Gene Expression in Skeletal Muscle of Exercised Rats.

We tested whether physiological doses of hydroxytyrosol (HT) may alter the mRNA transcription of key metabolic genes in exercised skeletal muscle. Two groups of exercise-trained Wistar rats, HTlow and HTmid, were supplemented with 0.31 and 4.61 mg/kg/d of HT, respectively, for 10 weeks. Another two groups of rats were not supplemented with HT; one remained sedentary and the other one was exercised. After the experimental period, the soleus muscle was removed for qRT-PCR and western blot analysis. The consumption of 4.61 mg/kg/d of HT during exercise increased the mRNA expression of important metabolic proteins. Specifically, 4.61 mg/kg/d of HT may upregulate long-chain fatty acid oxidation, lactate, and glucose oxidation as well as mitochondrial Krebs cycle in trained skeletal muscle. However, a 4.61 mg/kg/d of HT may alter protein translation, as in spite of the increment showed by CD36 and GLUT4 at the mRNA level this was not translated to higher protein content.

Hydroxytyrosol modifies skeletal muscle GLUT4/AKT/Rac1 axis in trained rats.

Training induces a number of healthy effects including a rise in skeletal muscle (SKM) glucose uptake. These adaptations are at least in part due to the reactive oxygen species produced within SKM, which is in agreement with the notion that antioxidant supplementation blunts some training-induced adaptations. Here, we tested whether hydroxytyrosol (HT), the main polyphenol of olive oil, would modify the molecular regulators of glucose uptake when HT is supplemented during exercise. Rats were included into sedentary and exercised (EXE) groups. EXE group was further divided into a group consuming a low HT dose (0.31 mg·kg·d; EXElow), a moderate HT dose (4.61 mg·kg·d; EXEmid), and a control group (EXE). EXE raised glucose transporter type 4 (GLUT4) protein content, Ras-related C3 botulinum toxin substrate 1 (Rac1) activity, and protein kinase b (AKT) phosphorylation in SKM. Furthermore, EXElow blunted GLUT4 protein content and AKT phosphorylation while EXEmid showed a downregulation of the GLUT4/AKT/Rac1 axis. Hence, a low-to-moderate dose of HT, when it is supplemented as an isolated compound, might alter the beneficial effect of training on basal AKT phosphorylation and Rac1 activity in rats.

Hydroxytyrosol influences exercise-induced mitochondrial respiratory complex assembly into supercomplexes in rats.

Hydroxytyrosol (HT) has been demonstrated to improve mitochondrial function, both in sedentary and in exercised animals. Herein, we assessed the effects of two different doses of HT on exercise-induced mitochondrial respiratory complex (C) assembly into supercomplexes (SCs) and the relation of the potential results to OPA1 levels and oxidative stress. Wistar rats were allocated into six groups: sedentary (SED), sedentary consuming 20 mg/kg/d of HT (SED-20), sedentary consuming 300 mg/kg/d of HT (SED-300); exercised (EXE), exercised consuming 20 mg/kg/d of HT (EXE-20) and exercised consuming 300 mg/kg/d of HT (EXE-300). Animals were exercised and/or supplemented for 10 weeks, and assembly of SCs, mitochondrial oxidative status and expression of OPA1 were quantified in the gastrocnemius muscle. Both EXE and EXE-20 animals exhibited increased assembly of CI into SCs, but this effect was absent in EXE-300 animals. Levels of CIII2 assembled into SCs were only increased in EXE-20 animals. Notably EXE-300 animals showed a decreased relative expression of s-OPA1 isoforms. Therefore, HT exerted dose-dependent effects on SC assembly in exercised animals. Although the mechanisms leading to SCs assembly in response to exercise and HT are unclear, it seems that a high HT dose can prevent SCs assembly during exercise by decreasing the expression of the s-OPA1 isoforms.

Effects of hydroxytyrosol dose on the redox status of exercised rats: the role of hydroxytyrosol in exercise performance.

Background: Hydroxytyrosol (HT) is a polyphenol found in olive oil that is known for its antioxidant effects. Here, we aimed to describe the effects of a low and high HT dose on the physical running capacity and redox state in both sedentary and exercised rats. Methods: Male Wistar rats were allocated into 6 groups: sedentary (SED; n = 10); SED consuming 20 mg/kg/d HT (SED20; n = 7); SED consuming 300 mg/kg/d HT (SED300; n = 7); exercised (EXE; n = 10); EXE consuming 20 mg/kg/d HT (EXE20; n = 10) and EXE consuming 300 mg/kg/d HT (EXE300; n = 10). All the interventions lasted 10 weeks; the maximal running velocity was assessed throughout the study, whereas daily physical work was monitored during each training session. At the end of the study, the rats were sacrificed by bleeding. Hemoglobin (HGB) and hematocrit (HCT) were measured in the terminal blood sample. Moreover, plasma hydroperoxide (HPx) concentrations were quantified as markers of lipid peroxidation. Results: In sedentary rats, HT induced an antioxidant effect in a dose-dependent manner without implications on running performance. However, if combined with exercise, the 300 mg/kg/d HT dosage exhibited a pro-oxidant effect in the EXE300 group compared with the EXE and EXE20 groups. The EXE20 rats showed a reduction in daily physical work and a lower maximal velocity than the EXE and EXE300 rats. The higher physical capacity exhibited by the EXE300 group was achieved despite the EXE300 rats expressing lower HGB levels and a lower HCT than the EXE20 rats. Conclusions: Our results suggest that a high HT dose induces a systemic pro-oxidant effect and may prevent the loss of performance that was observed with the low HT dose.