Lysates were prepared from cardiomyocytes that were plated on 35-mm plates and serum-starved for 24 h before exposure to testosterone for the indicated times. Glucose 10 mM was added to initiate glycolysis and measure glycolytic capacity in the cells. Inside the cell, the glucose phosphorylated by HK is metabolized to pyruvate. It has been shown that different pro-hypertrophic stimuli induce GLUT4 translocation and glucose uptake during cardiomyocyte growth 19, 20, 23. The increase in the rate of glycolysis from glucose involves modulation of glucose transport and positive regulation of key glycolytic enzymes .
B Glycolysis and maximal glycolytic capacity parameters induced by testosterone (100 nM) after 24 h of stimulation. Insulin (100 nM for 30 min) served as positive control of glucose uptake (Fig. 1c). A total of 12 male 8-week-old Sprague–Dawley rats were used, and orchiectomy (ORX) was performed in 8 rats to reduce circulating levels of testosterone. SiRNA-AMPKα2 (sc , Sta. Cruz Biotechnology) was used to decrease AMPK protein levels. Cells were cultured on gelatin-coated coverslips for 24 h, and then treated for 24 h with testosterone (100 nM), bicalutamide (2 µM) or CC (2 µM).
The regulatory effects of AMPK on metabolic and inflammatory processes were investigated in a letrozole-induced PCOS rat model . It is a combination of endocrine, metabolic, and reproductive disorders that lead to irregular menstrual cycles, dyslipidaemia, excessive body weight, oxidative stress, hyperandrogenism, and infertility 85,86. A previous study has shown that the transplantation of human amniotic mesenchymal stem cells (hAMSCs) in mice with age-related diminished ovarian reserves (AR-DORs) led to a significant improvement in ovarian function, while the apoptosis of granulosa and stromal cells in the ovaries was significantly reduced. In this study, follicle-stimulating hormone (FSH) was shown to stimulate mitophagy, reduce mitochondrial oedema, and increase the colocalisation of mitophagosomes with mitochondrial light chain 3 (LC3) through the activation of AMPK-PI3K/AKT pathways. In addition, AMPK signalling modulates hypothalamic fatty acid metabolism and brown adipose tissue thermogenesis, which contribute to hyperphagia and regulate energy balance during pregnancy in rats . AMPK also regulates the response to steroid hormones, the proliferation of epithelial cells and the receptivity of the uterus during implantation . AMPK expression is found throughout the female reproductive system and is important for the maintenance of female fertility.
Depolarized-induced suppression of excitation (DSE) is potentiated in POMC neurons from TP-treated animals and blocked by the CB1 receptor antagonist AM251. Collectively, this suggests that the ability of TP to further reduce excitatory synaptic input onto anorexigenic POMC neurons requires, at least in part, an AMPK-dependent increase in endocannabinoid tone that leads to a more extensive activation of presynaptic CB1 receptors, as summarized in Fig. To pharmacologically characterize the effects of CB1 receptor activation on mEPSC frequency, ARC neurons from vehicle- and TP-treated animals were tested with various concentrations of WIN 55,212-2 (0.01–3 μM). To this end, we conducted whole animal experiments in orchidectomized male guinea pigs to determine testosterone-induced changes in energy balance and AMPK activation in the ARC microdissected from hypothalamic slices. AMPK is an energy sensor that restores energy balance by activating processes that produce energy (e.g., lipid oxidation and glucose uptake) while inhibiting those that consume energy (e.g., protein synthesis) (28). Collectively, these results indicate that the androgen-induced increases in energy intake are mediated via an AMPK-dependent augmentation in endocannabinoid tone onto POMC neurons.
This activation restored the expression of proliferation-related molecules and proteins and reduced the inflammation-related protein NF-κB in senescent GCs. In another study, treatment with the plant flavonoid nobiletin (Nob) activated cell autophagy through AMPK and sirtuin-1 (SIRT1) in senescent SWF granulosa cells (SWF-GCs) induced by D-galactose. In addition, both the expression of AMPK and the ratio of phosphorylated FoxO3a to total FoxO3a were significantly increased, demonstrating the crucial role of the AMPK/FoxO3a signalling pathway in improving ovarian function . AMPK has been shown to regulate mitophagy and glycophagy in D-galactose (D-Gal)-induced senescent granulosa cells (GCs) in ageing chickens.
Several studies have demonstrated that metformin can reduce the levels of oxidative DNA damage and afford anti-oxidant protection (Attia et al., 2009; Onken and Driscoll, 2010; Martin-Montalvo et al., 2013). These functional markers include motility, capacitation, hyperactivation and the acrosome reaction (Hurtado de Llera et al., 2015). To traverse the female reproductive tract, it is essential that mammalian spermatozoa acquire the functional competence to achieve this objective in order to successfully fertilize the oocyte.
Two men in the testosterone group and 8 men in the placebo group dropped out and did not complete the study. The dose of testosterone was adjusted to keep serum free testosterone concentrations in the mid-normal range. Hypogonadal men were randomized to receive intramuscular injections of testosterone (therapy initiated with 200 mg) or placebo (saline 1 mL) every 2 weeks for 22 weeks.
The mean total testosterone concentrations in the testosterone and placebo groups at study end were 547 ± 234 and 280 ± 132 ng/dL, respectively. The mean free testosterone concentrations in the testosterone and placebo groups at study end were 12.0 ± 6.8 and 5.1 ± 1.7 ng/dL, respectively. Data are presented as mean ± SD, except for expression and phosphorylation of AMPKα, which are shown in graphs as mean ± SE.

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