AIM: Currently available medicines have little to offer in terms of supporting the regeneration of injured hepatic cells. Previous experimental studies have shown that resveratrol and metformin, less specific activators of AMP-activated protein kinase (AMPK) and sirtuin 1 (SIRT1), can effectively attenuate acute liver injury. The aim of this experimental study was to elucidate whether modulation of AMPK and SIRT1 activity can modify drug/paracetamol (APAP)-induced hepatocyte damage in vitro. METHODS: Primary rat hepatocytes were pretreated with mutual combinations of specific synthetic activators and inhibitors of SIRT1 and AMPK and followed by a toxic dose of APAP. At the end of cultivation, medium samples were collected for biochemical analysis of alanine-aminotransferase and nitrite levels. Hepatocyte viability, thiobarbituric reactive substances, SIRT1 and AMPK activity and protein expression were also assessed. RESULTS: The harmful effect of APAP was associated with decreased AMPK and SIRT1 activity and protein expression alongside enhanced oxidative stress in hepatocytes. The addition of AMPK activator (AICAR) or SIRT1 activator (CAY10591) significantly attenuated the deleterious effects of AMPK inhibitor (Compound C) on the hepatotoxicity of APAP. Furthermore, CAY10591 but not AICAR markedly decreased the deleterious effect of APAP in combination with SIRT1 inhibitor (EX-527). CONCLUSION: Our findings demonstrate that decreased AMPK activity is associated with the hepatotoxic effect of APAP which can be significantly attenuated by the administration of a SIRT1 activator. These findings suggest that differentiated modulation of AMPK and SIRT1 activity could therefore provide an interesting and novel therapeutic opportunity in the future to combat hepatocyte injury.

• Keywords
• 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), CAY10591, adenosine monophosphate protein kinase (AMPK), enzyme activation, hepatocyte protection, sirtuin 1 (SIRT1),
• MeSH
• Cyclopentanes pharmacology   MeSH
• Hepatocytes * metabolism   MeSH
• Rats MeSH
• Chemical and Drug Induced Liver Injury * etiology   genetics   metabolism   pathology   MeSH
• Acetaminophen toxicity   MeSH
• AMP-Activated Protein Kinases * chemistry   metabolism   pharmacology   MeSH
• Sirtuin 1 * metabolism   pharmacology   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• CAY10591 MeSH Browser
• Cyclopentanes MeSH
• Acetaminophen MeSH
• AMP-Activated Protein Kinases * MeSH
• Sirt1 protein, rat MeSH Browser
• Sirtuin 1 * MeSH

Two experiments were carried out to examine the impacts of hydroxytyrosol (HT) on lipid metabolism and mitochondrial function in Megalobrama amblycephala. Triplicate groups of fish were fed four test diets: (1) low-fat diet (LFD, 5% fat), (2) high-fat diet (HFD, 15% fat), (3) LFD + 100 mg/kg HT (LFD + HT), and (4) HFD + 100 mg/kg HT (HFD + HT) (in vivo). Hepatocytes from the same batch were exposed to three media including L-15 medium (L15), oleic acid (OA) medium [L15 + 400 μM OA], and OA + HT medium [L15 + 400 μM OA + 10 μM HT] to explore the roles of HT in mitochondrial function (in vitro). Fish fed HFD had excessive fat deposition in the liver, and HT inclusion in the HFD decreased hepatic fat deposition. Transmission electron microscopy revealed that the HFD triggers loss of cristae and metrical density and hydropic changes in mitochondria and that HT supplementation attenuates the ultrastructural alterations of mitochondria. The in vitro test showed that HT decreases fat deposition in hepatocytes, suppresses the reactive oxygen species formation, and facilitates the expression of phospho-AMPK protein and the genes involved in mitochondria biogenesis (PGC-1, NRF-1, TFAM) and autophagy (PINK1, Mul1, Atg5). These findings suggest the lipid-lowering effect of HT mediated by activation of mitochondrial biogenesis and autophagy through the AMPK pathway.

• Keywords
• blunt snout bream, fat deposition, hydroxytyrosol, mitochondrial biogenesis, mitophagy,
• MeSH
• Autophagy * MeSH
• Organelle Biogenesis MeSH
• Cyprinidae genetics   metabolism   MeSH
• Dietary Fats metabolism   MeSH
• Phenylethyl Alcohol analogs & derivatives   metabolism   MeSH
• Hepatocytes metabolism   MeSH
• Liver cytology   metabolism   MeSH
• Animal Feed analysis   MeSH
• Mitochondria metabolism   MeSH
• AMP-Activated Protein Kinases genetics   metabolism   MeSH
• Fish Proteins genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• 3,4-dihydroxyphenylethanol MeSH Browser
• Dietary Fats MeSH
• Phenylethyl Alcohol MeSH
• AMP-Activated Protein Kinases MeSH
• Fish Proteins MeSH

Metformin (MTF) is a widely used drug for the treatment of diabetes mellitus type 2 (DM2) and frequently used as an adjuvant therapy for polycystic ovarian syndrome, metabolic syndrome, and in some cases also tuberculosis. Its protective effect on the cardiovascular system has also been described. Recently, MTF was subjected to various analyzes and studies that showed its beneficial effects in cancer treatment such as reducing cancer cell proliferation, reducing tumor growth, inducing apoptosis, reducing cancer risk in diabetic patients, or reducing likelihood of relapse. One of the MTF's mechanisms of action is the activation of adenosine-monophosphate-activated protein kinase (AMPK). Several studies have shown that AMPK/mammalian target of rapamycin (mTOR) pathway has anticancer effect in vivo and in vitro. The aim of this review is to present the anticancer activity of MTF highlighting the importance of the AMPK/mTOR pathway in the cancer process.

• MeSH
• Antineoplastic Agents adverse effects   therapeutic use   MeSH
• Apoptosis drug effects   MeSH
• Hypoglycemic Agents therapeutic use   MeSH
• Humans MeSH
• Metformin adverse effects   therapeutic use   MeSH
• Neoplasms drug therapy   enzymology   pathology   MeSH
• Drug Repositioning MeSH
• AMP-Activated Protein Kinases metabolism   MeSH
• Signal Transduction MeSH
• TOR Serine-Threonine Kinases metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Antineoplastic Agents MeSH
• Hypoglycemic Agents MeSH
• Metformin MeSH
• MTOR protein, human MeSH Browser
• AMP-Activated Protein Kinases MeSH
• TOR Serine-Threonine Kinases MeSH

The importance of cellular metabolic adaptation in inducing robust T cell responses is well established. However, the mechanism by which T cells link information regarding nutrient supply to clonal expansion and effector function is still enigmatic. Herein, we report that the metabolic sensor adenosine monophosphate-activated protein kinase (AMPK) is a critical link between cellular energy demand and translational activity and, thus, orchestrates optimal expansion of T cells in vivo. AMPK deficiency did not affect T cell fate decision, activation, or T effector cell generation; however, the magnitude of T cell responses in murine in vivo models of T cell activation was markedly reduced. This impairment was global, as all T helper cell subsets were similarly sensitive to loss of AMPK which resulted in reduced T cell accumulation in peripheral organs and reduced disease severity in pathophysiologically as diverse models as T cell transfer colitis and allergic airway inflammation. T cell receptor repertoire analysis confirmed similar clonotype frequencies in different lymphoid organs, thereby supporting the concept of a quantitative impairment in clonal expansion rather than a skewed qualitative immune response. In line with these findings, in-depth metabolic analysis revealed a decrease in T cell oxidative metabolism, and gene set enrichment analysis indicated a major reduction in ribosomal biogenesis and mRNA translation in AMPK-deficient T cells. We, thus, provide evidence that through its interference with these delicate processes, AMPK orchestrates the quantitative, but not the qualitative, manifestation of primary T cell responses in vivo.

• Keywords
• AMPK, T cell, cellular metabolism, translation,
• MeSH
• Adenylate Kinase genetics   metabolism   MeSH
• Lymphocyte Activation MeSH
• Th17 Cells physiology   MeSH
• CD4-Positive T-Lymphocytes MeSH
• DNA-Binding Proteins genetics   metabolism   MeSH
• Adaptation, Physiological MeSH
• Colitis immunology   MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Mice, Knockout MeSH
• Mice MeSH
• Adoptive Transfer MeSH
• Gene Expression Regulation, Enzymologic MeSH
• T-Lymphocytes, Regulatory physiology   MeSH
• T-Lymphocytes, Helper-Inducer physiology   MeSH
• Th1 Cells physiology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Adenylate Kinase MeSH
• DNA-Binding Proteins MeSH
• RNA, Messenger MeSH
• Rag2 protein, mouse MeSH Browser

A large part of the world's population is affected by Alzheimer's disease (AD) and diabetes mellitus type 2, which cause both social and economic impacts. These two conditions are associated with one protein, AMPK. Studies have shown that vanadium complexes, such as bis(N',N'-dimethylbiguanidato)-oxovanadium(IV), VO(metf)2·H2O, are potential agents against AD. A crucial step in drug design studies is obtaining information about the structure and interaction of these complexes with the biological targets involved in the process through molecular dynamics (MD) simulations. However, MD simulations depend on the choice of a good force field that could present reliable results. Moreover, general force fields are not efficient for describing the properties of metal complexes, and a VO(metf)2·H2O-specific force field does not yet exist; thus, the proper development of a parameter set is necessary. Furthermore, this investigation is essential and relevant given the importance for both the scientific community and the population that is affected by this neurodegenerative disease. Therefore, the present work aims to develop and validate the AMBER force field parameters for VO(metf)2·H2O since the literature lacks such information on metal complexes and investigate through classical molecular dynamics the interactions made by the complex with the protein. The proposed force field proved to be effective for describing the vanadium complex (VC), supported by different analyses and validations. Moreover, it had a great performance when compared to the general AMBER force field. Beyond that, MD findings provided an in-depth perspective of vanadium complex-protein interactions that should be taken into consideration in future studies.

• MeSH
• Alzheimer Disease * drug therapy   MeSH
• Coordination Complexes * therapeutic use   MeSH
• Humans MeSH
• Neurodegenerative Diseases * MeSH
• AMP-Activated Protein Kinases chemistry   MeSH
• Molecular Dynamics Simulation MeSH
• Vanadium chemistry   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Coordination Complexes * MeSH
• AMP-Activated Protein Kinases MeSH
• Vanadium MeSH

Adiponectin is an adipokine increasing glucose and fatty acid metabolism and improving insulin sensitivity. The aim of this study was to investigate the role of adiponectin in the regulation of adipocyte lipolysis. Human adipocytes isolated from biopsies obtained during surgical operations from 16 non-obese and 17 obese subjects were incubated with 1) human adiponectin (20 microg/ml) or 2) 0.5 mM AICAR - activator of AMPK (adenosine monophosphate activated protein kinase). Following these incubations, isoprenaline was added (10(-6) M) to investigate the influence of adiponectin and AICAR on catecholamine-induced lipolysis. Glycerol concentration was measured as lipolysis marker. We observed that adiponectin suppressed spontaneous lipolysis by 21 % and isoprenaline-induced lipolysis by 14 % in non-obese subjects. These effects were not detectable in obese individuals, but statistically significant differences in the effect of adiponectin between obese and non-obese were not revealed by two way ANOVA test. The inhibitory effect of AICAR and adiponectin on lipolysis was reversed by Compound C. Our results suggest, that adiponectin in physiological concentrations inhibits spontaneous as well as catecholamine-induced lipolysis. This effect might be lower in obese individuals and this regulation seems to involve AMPK.

• MeSH
• Adiponectin pharmacology   MeSH
• Isoproterenol pharmacology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lipolysis drug effects   MeSH
• Obesity metabolism   MeSH
• AMP-Activated Protein Kinases metabolism   MeSH
• Adipocytes drug effects   enzymology   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Adiponectin MeSH
• ADIPOQ protein, human MeSH Browser
• Isoproterenol MeSH
• AMP-Activated Protein Kinases MeSH

Non-small cell lung cancer (NSCLC) results in high mortality and has gained increasing attention. C-Phycocyanin (C-PC) has been identified as a potential therapeutic inhibitor for NSCLC, but its underlying mechanism remains obscure. The gene expression of the long noncoding RNA neighbour of BRCAI RNA 2 (NBR2) in NSCLC cells was evaluated by quantitative reverse transcription-PCR. The cell capacity for proliferation and migration was examined by EdU and wound-healing assays. Furthermore, the viability and apoptosis of cells was measured with CCK-8 and annexin V/PI, respectively. Next, the protein level of activation of adenosine monophosphate- activated protein kinase and the rapamycin kinase (mTOR) signalling pathway-associated molecules was evaluated by western blotting. H292 cells were pre-treated with C-PC or transfected with plasmids encoding NBR2 or the shNBR2 plasmid, to over-express or knock down NBR2 expression, respectively. NBR2 expression was robustly down-regulated in NSCLC cell lines compared with a normal cell line (BEAS-2B). NBR2 over-expression inhibited migration and promoted apoptosis of H292 cells. Treatment of H292 cells with C-PC enhanced NBR2 levels in a dose- and time-dependent manner. Downregulation of NBR2 in H292 cells inhibited the activity of C-PC on cell proliferation, viability and clone formation. Further mechanistic investigation showed that the down-regulation of NBR2 abolished the modulatory effects of C-PC on the AMPK/mTOR signalling pathway. In conclusion, C-PC inhibits H292 cell growth by enhancing the NBR2/AMPK signalling pathway.

• MeSH
• Adenosine Monophosphate pharmacology   therapeutic use   MeSH
• Annexin A5 pharmacology   therapeutic use   MeSH
• Apoptosis MeSH
• Phycocyanin metabolism   pharmacology   therapeutic use   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Lung Neoplasms * drug therapy   genetics   metabolism   MeSH
• Carcinoma, Non-Small-Cell Lung * drug therapy   genetics   metabolism   MeSH
• Cell Movement MeSH
• Cell Proliferation MeSH
• AMP-Activated Protein Kinases genetics   metabolism   pharmacology   MeSH
• RNA, Long Noncoding * genetics   MeSH
• Sincalide metabolism   pharmacology   therapeutic use   MeSH
• Sirolimus pharmacology   therapeutic use   MeSH
• TOR Serine-Threonine Kinases metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Adenosine Monophosphate MeSH
• Annexin A5 MeSH
• Phycocyanin MeSH
• AMP-Activated Protein Kinases MeSH
• RNA, Long Noncoding * MeSH
• Sincalide MeSH
• Sirolimus MeSH
• TOR Serine-Threonine Kinases MeSH

Activation of autophagy suppresses ovarian cancer (OC). This in vitro study investigated whether the anti-tumour effect of exendin-4 against OC involves modulation of autophagy and figured out the possible mechanisms of action. SKOV-3 and OVCAR-3 cells (1 × 105/ml) were cultured in DMEM medium and treated with exendin-4 in the presence or absence of chloroquine (CQ), an autophagy inhibitor. In some cases, cells were also treated with exendin- 4 with or without pre-treatment with compound C (CC), an AMPK inhibitor, or insulin-like growth factor (IGF-1), a PI3K/Akt activator. Exendin-4 increased expression of beclin-1 and LC3I/II, suppressed expression of p62, reduced cell survival, migration, and invasion, and increased cell apoptosis and LDH release in both SKOV-3 and OVCAR-3 cells. Besides, exendin-4 reduced phosphorylation of mTORC1, 6SK, 4E-BP1, and Akt but increased phosphorylation of AMPK in both cell lines. These effects were associated with down-regulation of Bcl-2, suppression of nuclear phosphorylation of NF-κB p65, and increased expression of Bax and cleaved caspases 3/8. Chloroquine completely prevented the inhibitory effects of exendin-4 on the cell survival, Bcl-2, NF-κB, and cell invasiveness and abolished its stimulation of cell apoptosis and LDH release. Moreover, only the combined treatment with IGF-1 and CC completely abolished the observed effect of exendin-4 on the expression of beclin-1, LC3I/II, p62, as well as on cell survival, apoptosis, and LDH release. Exendin-4 exhibits a potent anti-tumour cytotoxic effect in SKOV-3 and OVCAR-3 cells by activating the markers of autophagy, mediated by activation of AMPK and inhibition of Akt.

• MeSH
• Apoptosis MeSH
• Autophagy MeSH
• Exenatide pharmacology   MeSH
• Phosphatidylinositol 3-Kinases MeSH
• Humans MeSH
• Mechanistic Target of Rapamycin Complex 1 MeSH
• Cell Line, Tumor MeSH
• Ovarian Neoplasms * drug therapy   MeSH
• AMP-Activated Protein Kinases MeSH
• Proto-Oncogene Proteins c-akt MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Exenatide MeSH
• Mechanistic Target of Rapamycin Complex 1 MeSH
• AMP-Activated Protein Kinases MeSH
• Proto-Oncogene Proteins c-akt MeSH

Atrial fibrillation is associated with atrial remodeling, in which connexin 43 (Cx43) and cell hypertrophy play important roles. In this study, apelin-13, an aliphatic peptide, was used to explore the protective effects of the adenosine monophosphate-activated protein kinase (AMPK)/mTOR signaling pathway on Cx43 expression and autophagy, using murine atrial HL-1 cells. The expression of Cx43, AMPK, B-type natriuretic peptide (BNP) and pathway-related proteins was detected by Western blot analysis. Cellular fluorescence imaging was used to visualize Cx43 distribution and the cytoskeleton. Our results showed that the Cx43 expression was significantly decreased in HL-1 cells treated with angiotensin II but increased in cells additionally treated with apelin-13. Meanwhile, apelin-13 decreased BNP expression and increased AMPK expression. However, the expression of Cx43 and LC3 increased by apelin-13 was inhibited by treatment with compound C, an AMPK inhibitor. In addition, rapamycin, an mTOR inhibitor, promoted the development of autophagy, further inhibited the protective effect on Cx43 expression and increased cell hypertrophy. Thus, apelin-13 enhances Cx43 expression and autophagy via the AMPK/mTOR signaling pathway, and serving as a potential therapeutic target for atrial fibrillation.

• MeSH
• Angiotensin II pharmacology   MeSH
• Autophagy drug effects   MeSH
• Cell Line MeSH
• Down-Regulation drug effects   MeSH
• Atrial Fibrillation metabolism   pathology   prevention & control   MeSH
• Myocytes, Cardiac drug effects   metabolism   pathology   MeSH
• Connexin 43 metabolism   MeSH
• Drug Interactions MeSH
• Intercellular Signaling Peptides and Proteins pharmacology   MeSH
• Mice MeSH
• AMP-Activated Protein Kinases metabolism   MeSH
• Signal Transduction drug effects   MeSH
• TOR Serine-Threonine Kinases metabolism   MeSH
• Vasoconstrictor Agents pharmacology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Angiotensin II MeSH
• apelin-13 peptide MeSH Browser
• GJA1 protein, mouse MeSH Browser
• Connexin 43 MeSH
• Intercellular Signaling Peptides and Proteins MeSH
• AMP-Activated Protein Kinases MeSH
• TOR Serine-Threonine Kinases MeSH
• Vasoconstrictor Agents MeSH

Responses of adipose cells to adrenoceptor regulation, including that of β-adrenoceptor (AR), and the signalling machinery involved in these responses are not sufficiently understood; information that is helpful for elucidating the adrenoceptor (adrenergic and β-AR)-responsive machinery is insufficient. We examined phospho-Thr-172 AMPK production in mouse-derived 3T3-L1 adipocytes treated with epinephrine or CL316243 (a β3-AR agonist) for 15 min. We also examined MAPK activation or G protein-associated PI3K activation or -associated PI3K p85 complex formation in rat epididymal (white) adipocytes treated with CL316243 for 15 min or aluminum fluoride (a G-protein signalling activator) for 20 min. Furthermore, we examined the effect of PTX (a trimeric G-protein inactivator) on p85 complex formation induced by aluminum fluoride treatment. Western blot analysis revealed that epinephrine or CL316243 treatment increased the phospho- Thr-172 AMPK (an active form of AMPK) level in 3T3-L1 adipocytes. Activated kinase analysis with a specific substrate showed that CL316243 or aluminum fluoride treatment activated MAPK in rat adipocytes. Immunoprecipitation experiments with a G-protein β subunit (Gβ) antibody showed that treatment of rat adipocytes with CL316243 activated PI3K and increased the PI3K p85 level in the Gβ antibody immunoprecipitates. Such an increase in the p85 level was similarly elicited by aluminum fluoride treatment in a PTX-sensitive manner. Our results provide possible clues for clarifying the signalling machinery involved in adrenoceptor responses, including those of β3-AR, in mouse-derived adipocytes and rat white adipocytes. Our findings advance the understanding of responses to adrenoceptor regulation in adipose cells and of the cellular signalling machinery present in the cells.

• MeSH
• Epinephrine pharmacology   MeSH
• Adipocytes, White enzymology   MeSH
• 3T3-L1 Cells MeSH
• Dioxoles pharmacology   MeSH
• Fluorides pharmacology   MeSH
• Phosphatidylinositol 3-Kinases metabolism   MeSH
• Phosphothreonine metabolism   MeSH
• Immunoprecipitation MeSH
• Rats MeSH
• Mitogen-Activated Protein Kinases metabolism   MeSH
• Mice MeSH
• Pertussis Toxin pharmacology   MeSH
• AMP-Activated Protein Kinases metabolism   MeSH
• GTP-Binding Proteins metabolism   MeSH
• Aluminum Compounds pharmacology   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Epinephrine MeSH
• aluminum fluoride MeSH Browser
• Dioxoles MeSH
• disodium (R,R)-5-(2-((2-(3-chlorophenyl)-2-hydroxyethyl)-amino)propyl)-1,3-benzodioxole-2,3-dicarboxylate MeSH Browser
• Fluorides MeSH
• Phosphatidylinositol 3-Kinases MeSH
• Phosphothreonine MeSH
• Mitogen-Activated Protein Kinases MeSH
• Pertussis Toxin MeSH
• AMP-Activated Protein Kinases MeSH
• GTP-Binding Proteins MeSH
• Aluminum Compounds MeSH