Role of the nitric oxide pathway in AMPK-mediated glucose uptake and GLUT4 translocation in heart muscle
|
|
- Barry Henderson
- 6 years ago
- Views:
Transcription
1 Am J Physiol Endocrinol Metab 287: E834 E841, First published July 20, 2004; doi: /ajpendo Role of the nitric oxide pathway in AMPK-mediated glucose uptake and GLUT4 translocation in heart muscle Ji Li, 1 Xiaoyue Hu, 1 Pradeepa Selvakumar, 1 Raymond R. Russell III, 1 Samuel W. Cushman, 2 Geoffrey D. Holman, 3 and Lawrence H. Young 1 1 Section of Cardiovascular Medicine, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520; 2 National Institutes of Health, Bethesda, Maryland 20892; and 3 Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom Submitted 1 June 2004; accepted in final form 28 June 2004 Li, Ji, Xiaoyue Hu, Pradeepa Selvakumar, Raymond R. Russell III, Samuel W. Cushman, Geoffrey D. Holman, and Lawrence H. Young. Role of the nitric oxide pathway in AMPK-mediated glucose uptake and GLUT4 translocation in heart muscle. Am J Physiol Endocrinol Metab 287: E834 E841, First published July 20, 2004; doi: /ajpendo AMP-activated protein kinase (AMPK) is a serine-threonine kinase that regulates cellular metabolism and has an essential role in activating glucose transport during hypoxia and ischemia. The mechanisms responsible for AMPK stimulation of glucose transport are uncertain, but may involve interaction with other signaling pathways or direct effects on GLUT vesicular trafficking. One potential downstream mediator of AMPK signaling is the nitric oxide pathway. The aim of this study was to examine the extent to which AMPK mediates glucose transport through activation of the nitric oxide (NO)-signaling pathway in isolated heart muscles. Incubation with 1 mm 5-amino-4-imidazole- 1- -carboxamide ribofuranoside (AICAR) activated AMPK (P 0.01) and stimulated glucose uptake (P 0.05) and translocation of the cardiomyocyte glucose transporter GLUT4 to the cell surface (P 0.05). AICAR treatment increased phosphorylation of endothelial NO synthase (enos) 1.8-fold (P 0.05). enos, but not neuronal NOS, coimmunoprecipitated with both the 2 and 1 AMPK catalytic subunits in heart muscle. NO donors also increased glucose uptake and GLUT4 translocation (P 0.05). Inhibition of NOS with N -nitro-l-arginine and N -methyl-l-arginine reduced AICAR-stimulated glucose uptake by 21 3% (P 0.05) and 25 4% (P 0.05), respectively. Inhibition of guanylate cyclase with ODQ and LY reduced AICAR-stimulated glucose uptake by 31 4% (P 0.05) and 22 3% (P 0.05), respectively, as well as GLUT4 translocation to the cell surface (P 0.05). Taken together, these results indicate that activation of the NO-guanylate cyclase pathway contributes to, but is not the sole mediator of, AMPK stimulation of glucose uptake and GLUT4 translocation in heart muscle. nitric oxide synthase; AMP-activated protein kinase; glucose transporter AMP-ACTIVATED PROTEIN KINASE (AMPK) is a serine-threonine kinase that has an important role in the regulation of cellular metabolism (14, 21), ion channels (13), and gene expression (44). AMPK is activated by increases in the AMP/ATP ratio and, as such, is a key signaling pathway during cellular metabolic or energetic stress. AMPK was initially found to be an important regulator of fatty acid oxidation in heart (23, 24) and skeletal muscle (43) but has also emerged as an important mediator of glucose metabolism (37). Specifically, AMPK activation by 5-amino-4-imidazole-1- -D-carboxamide ribofuranoside (AICAR) increases heart and skeletal muscle glucose uptake (15, 27, 36). AMPK also activates phosphofructokinase-2, which accelerates glycolysis (26). The importance of AMPK during hypoxic conditions is highlighted by recent findings that transgenic mice with deficient AMPK signaling have diminished glucose uptake in both the ischemic (35) and postischemic heart (35, 45) and hypoxic skeletal muscle (29). The mechanisms through which AMPK modulates glucose uptake are only partially understood and may involve both acute and chronic changes in glucose uptake. AMPK increases glucose transport by stimulating translocation of GLUT4 to the sarcolemma in heart (36) and skeletal muscle (25). In Clone 9 cells, AICAR increases GLUT1 activity without a change in content or distribution (1). Long-term AMPK activation with AICAR also increases the expression of GLUT4 in skeletal muscle (17). However, AMPK is not necessary for GLUT4 expression in muscle tissues, as transgenic mice with AMPK deficiency do not have reduced GLUT4 content (29, 35, 45). The downstream mechanisms through which AMPK mediates the acute activation of glucose transport remain uncertain. However, AMPK may potentially mediate its effect on glucose transport in part through interaction with the nitric oxide pathway. AMPK phosphorylates endothelial nitric oxide synthase (enos) on Ser 1177 (7, 8, 28), leading to NOS activation in a calcium-independent fashion. AMPK also phosphorylates muscle neuronal NOS (nnos) on Ser 1451 (7), although the functional significance of this interaction remains uncertain. In addition, NOS inhibition with N -nitro-l-arginine (L-NAME) reduces AICAR stimulation of glucose transport in skeletal muscle (12), and recent findings indicate that L-NAME also partially inhibits AICAR stimulation of muscle deoxyglucose uptake in the rat (40). However, the extent to which NOS modulates glucose uptake and GLUT translocation in heart muscle remains uncertain. Nitric oxide produced in endothelial cells may have an important paracrine role to modulate myocyte metabolism and function in muscle tissues. Nitric oxide has pleiotropic effects, depending in part on its concentration, and both the metabolic and hemodynamic states of the heart (39). In skeletal muscle from enos knockout mice, there is diminished insulin-stimulated glucose uptake, indicating that insulin activation of nitric oxide may contribute to the stimulation of glucose transport (5, 10, 42). At the present time, the role of the nitric oxide pathway in mediating AMPK s action in the heart is unknown. Address for reprint requests and other correspondence: L. H. Young, Section of Cardiovascular Medicine, TAC S460, Yale Univ. School of Medicine, New Haven, CT ( lawrence.young@yale.edu). E834 The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
2 The purpose of this study was to determine whether AMPK activation of heart glucose transport and GLUT4 translocation is mediated in part through interaction with and activation of NOS. An isolated rat left ventricular papillary muscle preparation was utilized to assess these effects independently of the hemodynamic and systemic consequences of NOS inhibition. The results indicate that AMPK binds to and phosphorylates enos in heart muscle and that nitric oxide and its downstream cgmp pathway modulate in part AMPK activation of glucose uptake and GLUT4 translocation in heart muscle. MATERIALS AND METHODS Animal preparationss. Male Sprague-Dawley rats weighing g were allowed to have standard chow and water ad libitum before experiments. All procedures were approved by the Yale University Animal Care and Use Committee. Rats were anesthetized by intraperitoneal injection of pentobarbital sodium (60 mg/kg). Anterior and posterior left ventricular papillary muscles (3 5 mg) were equilibrated in oxygenated phosphate-buffered saline (PBS) containing 1 mm MgCl 2, 1 mm CaCl 2, 5 mm glucose, and 1% BSA, as previously described (36). There was constant oxygen flow through the sealed muscle incubation containers, which were oscillated in a water bath at 37 C. Papillary muscles were then preincubated in buffer containing inhibitor, or inhibitor vehicle as control, for 30 min before the addition of pharmacological activators for min, as designated in RE- SULTS. NOS and guanylate cyclase inhibitors were used in optimally effective concentrations (33). Glucose transport. In experiments designed to assess glucose transport, papillary muscles were incubated as above, and 2-deoxy- [1-3 H]glucose (1 Ci/ml) was added during the final 30 min of incubation to measure the rates of glucose transport and phosphorylation. In addition, [U- 14 C]mannitol (0.1 Ci/ml) was added to measure the muscle extracellular space to correct for extracellular deoxyglucose (36). After incubations, muscles were washed in ice-cold PBS, blotted dry, weighed, solubilized, and counted by liquid scintillation (36). Glucose transporter surface labeling. Cell surface (sarcolemma and T tubule) glucose transporters were photoaffinity labeled with the cell-impermeant compound 4,4 -O-[2-[2-[2-[2-[2-[6 (biotinylamino)- hexanoyl]amino]ethoxy]ethoxy]ethoxy]-4-(1-azi-2,2,2,-trifluoroethyl)benzoyl]amino-1,3-propanediyl]bis-d-mannose (Bio-LC-ATB- BGPA), as previously described (14a, 38). In brief, after experimental incubations, muscles were labeled for 15 min in buffer containing 400 M Bio-LC-ATB-BGPA at 4 C and then irradiated with ultraviolet light (300 nm) twice for 3 min to cross-link the label with glucose transport proteins. Pooled (3 4) labeled muscles (10 20 mg wet wt) were homogenized in buffer containing 250 mm sucrose, 1 mm EDTA, 20 mm HEPES, and 1 g/ml of the proteinase inhibitors antipain, aprotinin, pepstatin, leupeptin, and 100 M 4-(2- aminoethyl)benzenesulfonyl fluoride (ph 7.2). Cell membranes were prepared by ultracentrifugation (227,000 g for 50 min at 4 C) and solubilized in PBS containing 2% Thesit and proteinase inhibitors. Membranes underwent centrifugation at 30,000 g for 30 min, and the solubilized membrane proteins were precipitated with streptavidinagarose (Pierce Chemical). The streptavidin-precipitated surfacelabeled proteins were washed several times and then subjected to SDS-PAGE and immunoblotting with GLUT4 or GLUT1 antibody. The surface-labeled GLUT1 and GLUT4 contents were normalized to the amount of these proteins in the total cell membrane fraction. AMPK activity. In experiments designed to assess AMPK activation, papillary muscles were frozen in liquid nitrogen, stored at 80 C, and subsequently homogenized in buffer containing 125 mm Tris, 1 mm EDTA, 1 mm EGTA, 250 mm mannitol, 50 mm sodium fluoride, 5 mm sodium pyrophosphate, 1 mm DTT, 1 mm benzamidine, 0.004% trypsin inhibitor, and 3 mm sodium azide, ph 7.5, at 4 C (9, 36). After centrifugation (13,200 g for 30 min), the supernatant underwent polyethylene glycol (PEG) precipitation, and the 2.5 6% fraction was used for measurement of total AMPK activity by use of the SAMS assay, as previously described (9, 36). Protein concentration was determined spectrophotometrically using the Bio- Rad reagent. The AMPK activity associated with specific catalytic subunits was examined following immunoprecipitation with polyclonal 1 and 2 antibodies, as previously described (9). AMPK-eNOS coimmunoprecipitation. After incubation with or without AICAR, heart muscles were homogenized in buffer containing 20 mm HEPES, 50 mm -glycerol phosphate, 2 mm EGTA, 1 mm DTT, 10 mm NaF, 1 mm sodium orthovanadate, 1% Triton X-100, 10% glycerol, 0.1% PMSF, 0.1 M leupeptin, and 10 ng/ml aprotinin at 4 C. After low-speed centrifugation, 400 g of protein were immunoprecipitated with 1 or 2 polyclonal antibodies, or nonimmune IgG as a negative control (9). After extensive washing, the immunoprecipitates were resuspended in Laemmli buffer for immunoblotting with enos or nnos antibodies. Immunoblotting. Proteins were combined with Laemmli sample buffer prior to SDS-PAGE. After transfer to PDVF membranes, proteins were immunoblotted, detected with enhanced chemiluminence, and quantified by densitometry of autoradiographs, as previously described (9). Immunoblots were performed with rabbit pan- ( 1/ 2) AMPK antibody at 1:10,000 dilution (kind gift from Dr. M. Birnbaum), sheep anti- 2 AMPK at 1:1,000 dilution (kind gift from Dr. D. G. Hardie), rabbit anti-pthr 172 AMPK antibody at 1:5,000 dilution (Cell Signaling), rabbit anti-pser 1177 enos (Cell Signaling) at 1:1,000 dilution, mouse anti-enos (BD Transduction Laboratory) at 1:2,500 dilution, rabbit anti-pser 1416 nnos (Upstate Biotechnology) at 1:2,000 dilution, and mouse anti-nnos (Santa Cruz Biotechnology) at 1:1,000 dilution. Statistical analysis. All data are reported as means SE. The number of experiments in each group is presented in the text, table, or figure legend. Data were analyzed by two-tailed, unpaired Student s t-test. Differences were considered significant at P RESULTS E835 Effects of AICAR on AMPK activation. We initially examined the effects of AICAR (1 mm) on AMPK activation in the isolated left ventricular papillary muscles. AICAR increased total AMPK activity threefold (P 0.01) in PEG-precipitated muscle homogenates (Fig. 1A). Immunoblots with anti-pthr 172 AMPK antibody showed that AICAR also increased AMPK catalytic subunit phosphorylation at its key regulatory site, reflecting phosphorylation by the upstream AMPK activating protein kinase (Fig. 1B). Finally, AICAR stimulated AMPK activity in both 1 (P 0.05) and 2 (P 0.01) immunoprecipitates in the heart muscles (Fig. 1C). Effects of AICAR on glucose transport. Incubation with AICAR stimulated heart muscle deoxyglucose transport (P 0.05; Fig. 2A), confirming our previous results (36). To determine whether this increase in glucose transport was due to translocation of glucose transporters, we assessed cell surface GLUT4 and GLUT1 content with the cell-impermeant biotinylated BGPA compound (38). AICAR increased cell surface GLUT4 (P 0.05) but did not affect the total cellular content of GLUT4, indicating that AMPK activation led to the glucose transporter translocation (Fig. 2B). AICAR also tended to increase cell surface GLUT1 (Fig. 2B). Effects of nitric oxide pathway activation on glucose uptake. To the extent that AMPK mediates its effect on glucose transport through interaction with the nitric oxide pathway, one would anticipate that nitric oxide donors would stimulate
3 E836 Fig Amino-4-imidazole-1- -D-carboxamide ribofuranoside (AICAR) activation of AMP-activated protein kinase (AMPK) A: AMPK activity. Total AMPK activity was assessed using the SAMS kinase assay in polyethylene glycol extracts of heart muscles incubated for 60 min with or without 1 mm AICAR (see MATERIALS AND METHODS). B: AMPK phosphorylation. AMPK catalytic subunit phosphorylation was assessed by immunoblotting with a phospho-thr 172 AMPK antibody. C: AMPK isoform kinase activity. The activity associated with the 1 or 2 catalytic AMPK subunits was measured after immunoprecipitation with isoform-specific antibodies. Values are means SE for 3 5 experiments, with 2 4 pooled heart muscles per experiment. P 0.05, *P 0.01 vs. control. glucose transport in heart muscle. Incubations with either sodium nitroprusside (SNP) or S-nitroso-N-acetyl-DL-penicillamine (SNAP) led to stimulation of glucose transport (P 0.05) at concentrations below 1 M (Fig. 3A). This was associated with an increase (P 0.05) in surface GLUT4, again indicating glucose transporter translocation (Fig. 3B). Nitric oxide mediates its downstream effects in part through activation of soluble guanylate cyclase and the formation of cgmp, which activates the downstream cgmp-dependent protein kinase pathway (11, 39). To assess whether cgmp might activate heart glucose transport, muscles were incubated with the cell-permeant cgmp analog bromo-cgmp. This compound was also found to increase (P 0.05) glucose uptake (Fig. 3C). Taken together, these results suggest that nitric oxide, or activation of its downstream cgmp pathway, leads to increased glucose uptake in heart muscle. AICAR activation and interaction with enos. Phosphorylation of enos at Ser 1177 by AMPK is known to increase its enzymatic activity (7, 8, 28). enos is highly expressed in heart capillary endothelial cells, but cardiac myocytes also contain small amounts of enos, nnos, and inducible NOS (inos) (30, 39). AMPK is also known to phosphorylate nnos, although the physiological consequences are uncertain (7, 12). Thus we evaluated whether AICAR stimulated phosphorylation of enos and/or nnos in heart muscles. AICAR incubation for 30 min increased enos phosphorylation at pser fold (P 0.05; Fig. 4A). To determine whether AMPK binds directly to enos in heart muscles, we immunoblotted 1 and 2 AMPK immunoprecipitates with enos antibodies and found coimmunoprecipitation of enos with both the 1 and 2 catalytic subunits but no evidence of enos in control nonimmune IgG immunoprecipitates (Fig. 4B). These findings were consistent with direct phosphorylation of enos by AMPK within heart muscle (8). Although small amounts of nnos were detectable in papillary muscles, there was no evidence that AICAR stimulated nnos phosphorylation at Ser 1416 or that there was binding of nnos to AMPK (Fig. 4, C and D). Effect of nitric oxide pathway inhibition on AICAR-stimulated glucose uptake. To determine the extent to which the nitric oxide pathway might contribute to the stimulation of heart muscle glucose transport by AICAR, muscles were incubated with NOS inhibitors before and during treatment with AICAR. Addition of 1 mm L-NAME (Fig. 5A) or 1 mm Fig. 2. Stimulation of glucose transport by AICAR. A: deoxyglucose transport. Isolated heart muscles were preincubated for min with or without 1 mm AICAR before addition of 2-deoxy-[1-3 H]glucose for an additional 30 min to measure glucose uptake (see MATERIALS AND METHODS). Values are means SE for 3 experiments, with 2 4 independently analyzed muscles per experiment. *P 0.05 vs. control. B: GLUT translocation. After control or AICAR incubations, cell surface GLUT4 (s-glut4) and GLUT1 (s-glut1) proteins were labeled with a cell-impermeant biotinylated bis-mannose compound (Bio-LC-ATB-BGPA) and cross-linked with UV irradiation (see MATERIALS AND METHODS). Labeled surface proteins were isolated on streptavidin-agarose and immunoblotted with GLUT4 or GLUT1 antibodies. Total cell membrane GLUT4 (t-glut4) or GLUT1 (t-glut 1) was measured before isolation with streptavidin. Representative immunoblots are shown, and ratios of surface to total GLUT4 and GLUT1 are expressed as means SE for 3 experiments, including 2 3 pooled muscles per experiment. *P 0.05 vs. control.
4 Fig. 3. Activation of glucose transport by nitric oxide donors. A: deoxyglucose transport. Isolated heart muscles were incubated without (control) or with the nitric oxide donors sodium nitroprusside (SNP) or S-nitroso-N-acetyl-DLpenicillamine (SNAP) ( M) for 60 min (see MATERIALS AND METHODS). Results indicate the means SE for 3 5 experiments, with 2 4 independently analyzed muscles per experiment. *P 0.05 vs. control. B: GLUT surface labeling. After control, SNP (1 M), or SNAP (1 M) incubations, surface GLUT4 and GLUT1 were labeled with Bio-LC-ATB-BGPA, isolated on streptavidin-agarose, and immunoblotted with specific GLUT4 or GLUT1 antibodies. Representative immunoblots are shown, and ratios of surface to total GLUT4 and GLUT1 are expressed as means SE for 3 experiments, including 2 3 pooled papillary muscles per experiment. *P 0.05 vs. control. C: bromo (Br)-cGMP and deoxyglucose transport. Heart muscles were incubated for 60 min with 10 M Br-cGMP, 1 M SNP, or 1 M SNAP (see MATERIALS AND METHODS). Results indicate means SE for 3 experiments, including 2 4 independently analyzed muscles per experiment. *P 0.05 vs. control. N -methyl-l-arginine (L-NMMA; Fig. 5B) inhibited AICARstimulated glucose uptake by 21 3% (P 0.05) or 25 4% (P 0.05), which is consistent with the hypothesis that NOS has a role in mediating the effect of AMPK to stimulate heart glucose transport. NOS inhibitors had no effect on baseline glucose transport (Fig. 5), suggesting that nitric oxide production is not required, at least acutely, to maintain baseline glucose uptake in heart muscles. To further distinguish whether AMPK activation of glucose transport is mediated through nitric oxide stimulation of guanylate cyclase or alternatively by non-cgmp-dependent mechanisms, we preincubated papillary muscles with the guanylate cyclase inhibitors 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1- one (ODQ, 25 M) or LY (10 M) before stimulation with AICAR. Both ODQ and LY inhibited AICARstimulated glucose uptake by 31 4% (P 0.05) and 22 3% (P 0.05), respectively (Fig. 6, A and C). In addition, ODQ also blunted AICAR-stimulated GLUT4 translocation using the cell surface-labeling technique (Fig. 6B), indicating that downstream activation of guanylate cyclase has a role in mediating the AMPK effect on cardiomyocyte glucose transport. Guanylate cyclase inhibitors had no effect on baseline glucose transport (Fig. 6, A and C), suggesting that cgmp production is not required to maintain baseline glucose uptake in heart muscles. DISCUSSION E837 The results of this study indicate that AMPK-mediated activation of the nitric oxide pathway plays a role in mediating the effects of AICAR to stimulate glucose transport and GLUT translocation in heart muscle. AICAR stimulated AMPK activity, which led to increased enos Ser 1177 phosphorylation, as well as to stimulation of heart glucose uptake and translocation of the cardiomyocyte glucose transporter GLUT4. Although AICAR may have additional nonspecific effects, we have previously shown that AICAR-stimulated glucose transport in heart muscles was not inhibited by either phosphatidylinositol 3-kinase inhibitors or adenosine receptor blockers (36). Low concentrations of nitric oxide donors, as well as bromo-cgmp, were found to increase heart muscle glucose uptake and cell surface GLUT4, indicating that the NOS-cGMP pathway activates heart glucose transporter translocation. This hypothesis was supported by the observation that either NOS or guanylate cyclase inhibitors partially blocked AICAR-stimulated (but not basal) glucose uptake. Although these findings demonstrate that NOS and downstream guanylate cyclase activation contribute to the effects of AMPK, there was 60 80% residual AICAR stimulation of glucose transport despite inhibition of the nitric oxide pathways, indicating that AMPK has additional important effects on glucose transport in heart muscle. These results expand our understanding of the mechanisms through which AMPK stimulates glucose transport in the heart and are of interest in light of recent studies showing that AMPK has an important role in the activation of glucose transport in the ischemic heart. Transgenic mice expressing dominant negative catalytic subunits of AMPK have a reduced glucose uptake response during both ischemia (35) and postischemic reperfusion (35, 45). AMPK deficiency also blocks hypoxia-stimulated glucose transport in skeletal muscle (29). AMPK is also known to be activated during exercise in both heart (9) and skeletal muscle (43), although it remains uncertain how important a role AMPK has in modulating glucose transport during exercise. In isolated skeletal muscle, contraction-stimulated glucose uptake is partially inhibited in AMPK-deficient dominant negative transgenic mice (29), but appears to be normal in muscles from 2 knockout mice (19).
5 E838 Fig. 4. AMPK activation of endothelial nitric oxide synthase (enos). A: enos phosphorylation. Isolated heart papillary muscles were incubated in the presence or absence of AICAR (1 mm) for 30 min and homogenates immunoblotted with antibodies to pser 1177 enos and total enos. Representative immunoblots are shown, and ratios of p-enos to total enos are expressed as means SE for 3 experiments, including 2 3 pooled muscles per experiment. *P 0.05 vs. control. B: AMPK binding to enos. Muscles were incubated in the presence or absence of AICAR (1 mm) for 30 min and homogenates immunoprecipitated (IP) with antibodies to 1 or 2 AMPK or nonimmune sheep IgG. Immunoprecipitates were immunoblotted with antibodies to enos, as well as with antibodies to 1 and 2 AMPK to determine recovery. C. neuronal (n)nos phosphorylation. Isolated heart muscles were incubated in the presence or absence of AICAR (1 mm) for 30 min and homogenates immunoblotted with antibodies to pser 1416 nnos and total nnos (rat brain homogenate was used as a positive control). D: nnos binding. Heart 1 or 2 immunoprecipitates were immunoblotted with antibodies to nnos, as well as with antibodies to 1 and 2 AMPK to determine recovery. Fig. 5. NOS inhibition and AICAR-stimulated deoxyglucose transport. Isolated heart muscles were preincubated for 30 min with 1 mm L-NAME (A) or L-NMMA (B) before incubation with or without AICAR (1 mm) for 60 min. 2-Deoxy-[1-3 H]glucose was added during the last 30 min to measure glucose uptake (see MATERIALS AND METHODS). Results indicate means SE for 3 experiments, including 2 4 independently analyzed papillary muscles per experiment. *P 0.01 vs. control, P 0.05 vs. control, P 0.05 vs. AICAR. These latter findings suggest that additional redundant signaling pathways may exist that compensate for AMPK deficiency during contraction. Whether such mechanisms are simply recruited in transgenic mice as the result of developmental and/or chronic AMPK deficiency, or would be operative in normal muscle tissue with acute AMPK inhibition, remains uncertain. The present results support the hypothesis that activation of the nitric oxide-guanylate cyclase pathway plays a role in mediating the effects of AMPK on glucose transport in heart muscle. These experiments were performed in isolated left ventricular papillary muscles, which contain a variety of cell types, including cardiac myocytes and endothelial cells. However, cardiac myocytes are the only cells in the heart which contain GLUT4 (47) and account for the bulk of glucose transport in heart because they also predominate by mass (46). Thus, taken together, the results from GLUT4 surface labeling and glucose uptake experiments indicate that the enos-guanylate cyclase pathway impacts on cardiomyocyte glucose uptake during AICAR stimulation. We observed not only that AICAR increased enos Ser 1177 phosphorylation but also that AMPK coimmunoprecipitated with enos, suggesting that AMPK directly activated enos. These findings are consistent with previous reports in intact heart and skeletal muscle (8, 41). Experiments in isolated mouse H-2Kb muscle cells (12) raised the possibility that AMPK interacts directly with enos in muscle cells. However, the current experiments, as well as those in heart and skeletal muscle (8, 41), do not directly define the extent to which AMPK activation of enos occurs in the endothelial cells, endocardial cells, or myocytes in the intact muscle tissue.
6 Fig. 6. Guanylate cyclase inhibition and AICAR-stimulated deoxyglucose uptake. Isolated heart muscles were preincubated for 30 min with 25 M ODQ (A) or 10 M LY (C), before incubation with or without AICAR (1 mm) for 60 min (see MATERIALS AND METHODS). Results indicate means SE for 3 experiments, including 3 4 independently analyzed muscles per experiment. *P 0.01 vs. control, P 0.05 vs. control, P 0.05 vs. AICAR. B: guanylate cyclase inhibiton and AICAR-stimulated GLUT4 translocation. After control or AICAR incubation with or without ODQ (25 M), cell surface GLUT4 was labeled with Bio-LC-ATB-BGPA, isolated on streptavidin-agarose, and immunoblotted with specific GLUT4 antibodies. Representative immunoblots are shown, and ratios of surface to total GLUT4 are expressed as means SE for 3 experiments, including 2 3 pooled muscles per experiment. *P 0.05 vs. control, P 0.05 vs. AICAR. Vascular endothelial cell enos is known to be activated by AICAR (18), and endothelial cells predominantly express the 1 isoform of AMPK (28). It is interesting in this regard that we found that AICAR activated the 1 isoform and bound to enos in the heart muscles, suggesting that AICAR activation of endothelial cell AMPK and enos may have an important role in the stimulation of cardiomyocyte glucose transport through a paracrine mechanism. However, additional immunoprecipitation experiments demonstrated that enos was also associated with 2 AMPK, the more predominant isoform in cardiac myocytes, which is virtually absent from endothelial cells (28). Although cardiac myocyte expression of enos is generally low, there is heterogeneity within the heart with greater enos expression in specialized endocardial cells that line the cardiac chambers (6, 30) and epicardial cardiomyocytes (4). Thus these findings suggest that both autocrine and paracrine mechanisms may be involved to some extent in the interaction between the AMPK, enos, and glucose transport pathways in heart muscle. Heart and skeletal muscle cells also express nnos (7, 12, 30, 39). In skeletal muscle, exercise activates AMPK and E839 increases the phosphorylation of the muscle isoform of nnos (nnos-mu) on pser 1451 (7). The physiological effect of nnos phosphorylation in skeletal muscle remains uncertain. We found a small amount of nnos present in the heart papillary muscles, but there was no evidence that AICAR treatment increased nnos phosphorylation or that nnos was bound to either the 1 or 2 isoform of AMPK. Thus the results of these experiments suggest that AMPK interaction with the nitric oxide-cgmp pathway involves modulation of enos, rather than nnos, in heart muscle. Nitric oxide stimulates glucose transport in isolated skeletal muscle (2, 48), where it is thought to have a role in mediating both insulin- (20, 22) and exercise-stimulated glucose uptake (34). In skeletal muscle and other tissues from enos knockout mice, there is diminished insulin-stimulated glucose uptake, indicating that insulin activation of NOS may contribute to the stimulation of glucose transport (5, 10, 42). In this study, we observed that relatively low concentrations of nitric oxide donors (0.1 M) increased glucose uptake and stimulated GLUT4 translocation in isolated heart papillary muscles. The effects of nitric oxide on heart GLUT4 translocation have not been previously examined. Prior studies in working hearts have suggested that nitric oxide may activate fatty acid metabolism and inhibit overall glucose utilization (32), leading to the postulate that deficient enos activity in heart failure may reduce fatty acid oxidation and increase glucose metabolism (31). However, nitric oxide modulates cardiac contractility and oxidative metabolism in vivo, which may counterbalance the direct effects of nitric oxide to activate glucose transport. In the present experiments, the utilization of an isolated, quiescent papillary muscle preparation enabled us to examine more directly the role of nitric oxide and NOS inhibition on glucose transport and GLUT4 translocation. Although increased nitric oxide activates GLUT4 translocation, NOS inhibition experiments had no effect on basal glucose uptake, indicating that nitric oxide is not required to maintain basal glucose uptake in isolated heart muscles. The findings that NOS inhibitors decrease AICAR-stimulated glucose transport in heart muscles are consistent overall with findings in skeletal muscle-derived mouse H-2Kb cells and isolated rat skeletal muscles (12), as well as in rat skeletal muscle in vivo (40). In isolated skeletal muscles, there was virtually complete inhibition of AICAR-stimulated glucose uptake by NOS inhibitors (12). In recent in vivo studies, L-NAME decreased AICAR-stimulated deoxyglucose uptake but also decreased basal uptake (40). There was partial (20 40%) inhibition of AICAR-stimulated glucose transport by NOS inhibitors in the present study in heart muscles. These results indicate that the majority of AMPK-mediated glucose transport in the heart is mediated either through additional downstream signaling pathways or by the direct effects of AMPK on GLUT4 vesicular trafficking. Although additional pathways that might mediate AMPK s effects on glucose transport in heart are unknown, the phosphatidylinositol-3- kinase pathway, which plays an essential role in insulin signaling, does not appear to be involved (3, 16, 36). In addition, the partial inhibition of glucose transport and GLUT4 translocation seen with guanylate cyclase inhibitors is also consistent with prior studies in skeletal muscle cells (12). These findings indicate that guanylate cyclase is an important downstream mediator of glucose transport activation by nitric oxide, al-
7 E840 though the mechanism by which cgmp activates glucose transport in muscle remains uncertain. Thus these findings indicate that the nitric oxide-guanylate cyclase pathway partially contributes to the AMPK stimulation of glucose transport in heart muscle. The importance of this pathway in mediating the activation of glucose transport in the heart during exercise and ischemia is of interest for future studies. ACKNOWLEDGMENTS We express appreciation to Dr. William Sessa for helpful suggestions. GRANTS This work was supported by United States Public Health Service (National Heart, Lung, and Blood Institute) Grant RO1-HL (L. H. Young) and a Medical Student Research fellowship from the Heritage Affiliate of the American Heart Association (P. Selvakumar). REFERENCES 1. Abbud W, Habinowski S, Zhang JZ, Kendrew J, Elkairi FS, Kemp BE, Witters LA, and Ismail-Beigi F. Stimulation of AMP-activated protein kinase (AMPK) is associated with enhancement of Glut1-mediated glucose transport. Arch Biochem Biophys 380: , Balon TW and Nadler JL. Nitric oxide mediates skeletal glucose transport. Am J Physiol Endocrinol Metab 270: E258 E259, Bergeron R, Russell RR, Young LH, Ren JM, Marcucci M, Lee A, and Shulman GI. Effect of AMPK activation on muscle glucose metabolism in conscious rats. Am J Physiol Endocrinol Metab 276: E938 E944, Brahmajothi MV and Campbell DL. Heterogeneous basal expression of nitric oxide synthase and superoxide dismutase isoforms in mammalian heart : implications for mechanisms governing indirect and direct nitric oxide-related effects. Circ Res 85: , Browne SE, Ayata C, Huang PL, Moskowitz MA, and Beal MF. The cerebral metabolic consequences of nitric oxide synthase deficiency: glucose utilization in endothelial and neuronal nitric oxide synthase null mice. J Cereb Blood Flow Metab 19: , Brutsaert DL. Cardiac endothelial-myocardial signaling: its role in cardiac growth, contractile performance, and rhythmicity. Physiol Rev 83: , Chen ZP, McConell GK, Michell BJ, Snow RJ, Canny BJ, and Kemp BE. AMPK signaling in contracting human skeletal muscle: acetyl-coa carboxylase and NO synthase phosphorylation. Am J Physiol Endocrinol Metab 279: E1202 E1206, Chen ZP, Mitchelhill KI, Michell BJ, Stapleton D, Rodriguez-Crespo I, Witters LA, Power DA, Ortiz de Montellano PR, and Kemp BE. AMP-activated protein kinase phosphorylation of endothelial NO synthase. FEBS Lett 443: , Coven DL, Hu X, Cong L, Bergeron R, Shulman GI, Hardie DG, and Young LH. Physiological role of AMP-activated protein kinase in the heart: graded activation during exercise. Am J Physiol Endocrinol Metab 285: E629 E636, Duplain H, Burcelin R, Sartori C, Cook S, Egli M, Lepori M, Vollenweider P, Pedrazzini T, Nicod P, Thorens B, and Scherrer U. Insulin resistance, hyperlipidemia, and hypertension in mice lacking endothelial nitric oxide synthase. Circulation 104: , Friebe A and Koesling D. Regulation of nitric oxide-sensitive guanylyl cyclase. Circ Res 93: , Fryer LG, Hajduch E, Rencurel F, Salt IP, Hundal HS, Hardie DG, and Carling D. Activation of glucose transport by AMP-activated protein kinase via stimulation of nitric oxide synthase. Diabetes 49: , Hallows KR, McCane JE, Kemp BE, Witters LA, and Foskett JK. Regulation of channel gating by AMP-activated protein kinase modulates cystic fibrosis transmembrane conductance regulator activity in lung submucosal cells. J Biol Chem 278: , Hardie DG, Scott JW, Pan DA, and Hudson ER. Management of cellular energy by the AMP-activated protein kinase system. FEBS Lett 546: , a. Hashimoto M, Hatanaka Y, Yang J, Dhesi J, and Holman GD. Synthesis of biotinylated bis(d-glucose) derivatives for glucose transporter photoaffinity labelling. Carbohydr Res 331: , Hayashi T, Hirshman MF, Fujii N, Habinowski SA, Witters LA, and Goodyear LJ. Metabolic stress and altered glucose transport: activation of AMP-activated protein kinase as a unifying coupling mechanism. Diabetes 49: , Hayashi T, Hirshman MF, Kurth EJ, Winder WW, and Goodyear LJ. Evidence for 5 AMP-activated protein kinase mediation of the effect of muscle contraction on glucose transport. Diabetes 47: , Holmes BF, Kurth-Kraczek EJ, and Winder WW. Chronic activation of 5 -AMP-activated protein kinase increases GLUT-4, hexokinase, and glycogen in muscle. J Appl Physiol 87: , Ido Y, Carling D, and Ruderman N. Hyperglycemia-induced apoptosis in human umbilical vein endothelial cells: inhibition by the AMP-activated protein kinase activation. Diabetes 51: , Jorgensen SB, Viollet B, Andreelli F, Frosig C, Birk JB, Schjerling P, Vaulont S, Richter EA, and Wojtaszewski JF. Knockout of the 2 but not 1 5 -AMP-activated protein kinase isoform abolishes 5-aminoimidazole-4-carboxamide ribofuranoside but not contraction-induced glucose uptake in skeletal muscle. J Biol Chem 279: , Kapur S, Bédard S, Marcotte B, Côté C, and Marette A. Expression of nitric oxide synthase in skeletal muscle: a novel role for nitric oxide as a modulator of insulin action. Diabetes 46: , Kemp BE, Stapleton D, Campbell DJ, Chen ZP, Murthy S, Walter M, Gupta A, Adams JJ, Katsis F, Van Denderen B, Jennings IG, Iseli T, Michell BJ, and Witters LA. AMP-activated protein kinase, super metabolic regulator. Biochem Soc Trans 31: , Kingwell BA, Formosa M, Muhlmann M, Bradley SJ, and McConell GK. Nitric oxide synthase inhibition reduces glucose uptake during exercise in individuals with type 2 diabetes more than in control subjects. Diabetes 51: , Kudo N, Barr AJ, Barr RL, Desai S, and Lopaschuk GD. High rates of fatty acid oxidation during reperfusion of ischemic hearts are associated with a decrease in malonyl-coa levels due to an increase in 5 -AMPactivated protein kinase inhibition of acetyl-coa carboxylase. J Biol Chem 270: , Kudo N, Gillespie JG, Kung L, Witters LA, Schulz R, Clanachan AS, and Lopaschuk GD. Characterization of 5 AMP-activated protein kinase activity in the heart and its role in inhibiting acetyl-coa carboxylase during reperfusion following ischemia. Biochim Biophys Acta 1301: 67 75, Kurth-Kraczek EJ, Hirshman MF, Goodyear LJ, and Winder WW. 5 AMP-activated protein kinase activation causes GLUT4 translocation in skeletal muscle. Diabetes 48: , Marsin AS, Bertrand L, Rider MH, Deprez J, Beauloye C, Vincent MF, Van den Berghe G, Carling D, and Hue L. Phosphorylation and activation of heart PFK-2 by AMPK has a role in the stimulation of glycolysis during ischaemia. Curr Biol 10: , Merrill GF, Kurth EJ, Hardie DG, and Winder WW. AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle. Am J Physiol Endocrinol Metab 273: E1107 E1112, Morrow VA, Foufelle F, Connell JM, Petrie JR, Gould GW, and Salt IP. Direct activation of AMP-activated protein kinase stimulates nitricoxide synthesis in human aortic endothelial cells. J Biol Chem 278: , Mu J, Brozinick JT Jr, Valladares O, Bucan M, and Birnbaum MJ. A role for AMP-activated protein kinase in contraction- and hypoxia-regulated glucose transport in skeletal muscle. Mol Cell 7: , Mungrue IN, Husain M, and Stewart DJ. The role of NOS in heart failure: lessons from murine genetic models. Heart Fail Rev 7: , Recchia FA, McConnell PI, Bernstein RD, Vogel TR, Xu X, and Hintze TH. Reduced nitric oxide production and altered myocardial metabolism during the decompensation of pacing-induced heart failure in the conscious dog. Circ Res 83: , Recchia FA, Osorio JC, Chandler MP, Xu X, Panchal AR, Lopaschuk GD, Hintze TH, and Stanley WC. Reduced synthesis of NO causes marked alterations in myocardial substrate metabolism in conscious dogs. Am J Physiol Endocrinol Metab 282: E197 E206, Rees DD, Palmer RM, Schulz R, Hodson HF, and Moncada S. Characterization of three inhibitors of endothelial nitric oxide synthase in vitro and in vivo. Br J Pharmacol 101: , Roberts CK, Barnard RJ, Scheck SH, and Balon TW. Exercisestimulated glucose transport in skeletal muscle is nitric oxide dependent. Am J Physiol Endocrinol Metab 273: E220 E225, 1997.
8 E Russell RR, Li J, Coven DL, Pypaert M, Zechner C, Palmieri M, Giordano FJ, Mu J, Birnbaum MJ, and Young LH. AMP-activated protein kinase mediates ischemic glucose uptake and prevents post-ischemic cardiac dysfunction, apoptosis and injury. J Clin Invest 114: , Russell RR, Bergeron R, Shulman GI, and Young LH. Translocation of myocardial GLUT4 and increased glucose uptake through activation of AMPK by AICAR. Am J Physiol Heart Circ Physiol 277: H643 H649, Rutter GA, Da Silva Xavier G., and Leclerc I. Roles of 5 -AMPactivated protein kinase (AMPK) in mammalian glucose homoeostasis. Biochem J 375: 1 16, Ryder JW, Yang J, Galuska D, Rincon J, Bjornholm M, Krook A, Lund S, Pedersen O, Wallberg-Henriksson H, Zierath JR, and Holman GD. Use of a novel impermeable biotinylated photolabeling reagent to assess insulin- and hypoxia-stimulated cell surface GLUT4 content in skeletal muscle from type 2 diabetic patients. Diabetes 49: , Schulz R, Kelm M, and Heusch G. Nitric oxide in myocardial ischemia/ reperfusion injury. Cardiovasc Res 61: , Shearer J, Fueger PT, Vorndick B, Bracy DP, Rottman JN, Clanton JA, and Wasserman DH. AMP kinase-induced skeletal muscle glucose but not long-chain fatty acid uptake is dependent on nitric oxide. Diabetes 53: , Stephens TJ, Chen ZP, Canny BJ, Michell BJ, Kemp BE, and Mc- Conell GK. Progressive increase in human skeletal muscle AMPK 2 activity and ACC phosphorylation during exercise. Am J Physiol Endocrinol Metab 282: E688 E694, Vicent D, Ilany J, Kondo T, Naruse K, Fisher SJ, Kisanuki YY, Bursell S, Yanagisawa M, King GL, and Kahn CR. The role of endothelial insulin signaling in the regulation of vascular tone and insulin resistance. J Clin Invest 111: , Winder WW and Hardie DG. Inactivation of acetyl-coa carboxylase and activation of AMP-activated protein kinase in muscle during exercise. Am J Physiol Endocrinol Metab 270: E299 E304, Woods A, Azzout-Marniche D, Foretz M, Stein SC, Lemarchand P, Ferre P, Foufelle F, and Carling D. Characterization of the role of AMP-activated protein kinase in the regulation of glucose-activated gene expression using constitutively active and dominant negative forms of the kinase. Mol Cell Biol 20: , Xing Y, Musi N, Fujii N, Zou L, Luptak I, Hirshman MF, Goodyear LJ, and Tian R. Glucose metabolism and energy homeostasis in mouse hearts overexpressing dominant negative alpha2 subunit of AMP-activated protein kinase. J Biol Chem 278: , Young LH, Coven DL, and Russell RR III. Cellular and molecular regulation of cardiac glucose transport. J Nucl Cardiol 7: , Young LH, Renfu Y, Russell RR, Hu X, Caplan MJ, Ren J, Shulman GI, and Sinusas AJ. Low-flow ischemia leads to translocation of canine heart GLUT-4 and GLUT-1 glucose transporters to the sarcolemma in vivo. Circulation 95: , Young ME, Radda GK, and Leighton B. Nitric oxide stimulates glucose transport and metabolism in rat skeletal muscle in vitro. Biochem J 322: , Downloaded from by on November 23, 2017
Does Nitric Oxide Regulate Skeletal Muscle Glucose Uptake during Exercise?
ARTICLE Does Nitric Oxide Regulate Skeletal Muscle Glucose Uptake during Exercise? Glenn K. McConell 1 and Bronwyn A. Kingwell 2 1 Department of Physiology, The University of Melbourne, Parkville, Australia;
More informationPhysiological role of AMP-activated protein kinase in the heart: graded activation during exercise
Am J Physiol Endocrinol Metab 285: E629 E636, 2003. First published May 20, 2003; 10.1152/ajpendo.00171.2003. Physiological role of AMP-activated protein kinase in the heart: graded activation during exercise
More informationAMP-Activated Protein Kinase Activates p38 Mitogen-Activated Protein Kinase by Increasing Recruitment of p38 MAPK to TAB1 in the Ischemic Heart
AMP-Activated Protein Kinase Activates p38 Mitogen-Activated Protein Kinase by Increasing Recruitment of p38 MAPK to TAB1 in the Ischemic Heart Ji Li, Edward J. Miller, Jun Ninomiya-Tsuji, Raymond R. Russell
More informationRegulation of glucose transport by the AMP-activated protein kinase
Proceedings of the Nutrition Society (2004), 63, 205 210 g The Authors 2004 DOI:10.1079/PNS2004340 The 12th Conference of the International Research Group on the Biochemistry of Exercise was held at Maastricht
More informationCritical Review. Skeletal Muscle Glucose Uptake During Exercise: A Focus on Reactive Oxygen Species and Nitric Oxide Signaling
IUBMB Life, 61(5): 479 484, May 2009 Critical Review Skeletal Muscle Glucose Uptake During Exercise: A Focus on Reactive Oxygen Species and Nitric Oxide Signaling Troy L. Merry and Glenn K. McConell Department
More informationIncreased GLUT-4 translocation mediates enhanced insulin sensitivity of muscle glucose transport after exercise
Increased GLUT-4 translocation mediates enhanced insulin sensitivity of muscle glucose transport after exercise POLLY A. HANSEN, LORRAINE A. NOLTE, MAY M. CHEN, AND JOHN O. HOLLOSZY Department of Medicine,
More informationCirc Res. 2005;96: ; originally published online January 13, 2005; doi: /01.RES d2
Dual Mechanisms Regulating AMPK Kinase Action in the Ischemic Heart Suzanne J. Baron, Ji Li, Raymond R. Russell III, Dietbert Neumann, Edward J. Miller, Roland Tuerk, Theo Wallimann, Rebecca L. Hurley,
More informationThe AMP-activated protein kinase activator AICAR does not induce GLUT4 translocation to transverse tubules. protein kinases and in skeletal muscle
The AMP-activated protein kinase activator AICAR does not induce GLUT4 translocation to transverse tubules but stimulates glucose uptake and p38 mitogenactivated protein kinases and in skeletal muscle
More informationExercise and insulin stimulate glucose transport
Ca 2 and AMPK Both Mediate Stimulation of Glucose Transport by Muscle Contractions David C. Wright, Kathleen A. Hucker, John O. Holloszy, and Dong Ho Han It is now generally accepted that activation of
More informationActivation of AMPK - and -isoform complexes in the intact ischemic rat heart
Am J Physiol Heart Circ Physiol 291: H1927 H1934, 2006. First published April 28, 2006; doi:10.1152/ajpheart.00251.2006. Activation of AMPK - and -isoform complexes in the intact ischemic rat heart Ji
More informationMedical Biochemistry and Molecular Biology department
Medical Biochemistry and Molecular Biology department Cardiac Fuels [Sources of energy for the Cardiac muscle] Intended learning outcomes of the lecture: By the end of this lecture you would be able to:-
More informationActivation of AMPK is essential for AICAR-induced glucose uptake by skeletal muscle but not adipocytes
Am J Physiol Endocrinol Metab 282: E1239 E1244, 2002. First published February 19, 2002; 10.1152/ajpendo.00455.2001. Activation of AMPK is essential for AICAR-induced glucose uptake by skeletal muscle
More informationMetabolism of cardiac muscle. Dr. Mamoun Ahram Cardiovascular system, 2013
Metabolism of cardiac muscle Dr. Mamoun Ahram Cardiovascular system, 2013 References This lecture Mark s Basic Medical Biochemistry, 4 th ed., p. 890-891 Hand-out Why is this topic important? Heart failure
More informationAMP-activated protein kinase mediates ischemic glucose uptake and prevents postischemic cardiac dysfunction, apoptosis, and injury
Related Commentary, page 465 Research article AMP-activated protein kinase mediates ischemic glucose uptake and prevents postischemic cardiac dysfunction, apoptosis, and injury Raymond R. Russell III,
More informationChronic activation of 5 -AMP-activated protein kinase increases GLUT-4, hexokinase, and glycogen in muscle
highlighted topics Chronic activation of 5 -AMP-activated protein kinase increases GLUT-4, hexokinase, and glycogen in muscle B. F. HOLMES, E. J. KURTH-KRACZEK, AND W. W. WINDER Department of Zoology,
More informationCritical Role of 5 -AMP-activated Protein Kinase in the Stimulation of Glucose. Transport in Response to Inhibition of Oxidative Phosphorylation
Page 1 of 56 Articles in PresS. Am J Physiol Cell Physiol (August 30, 2006). doi:10.1152/ajpcell.00196.2006 Critical Role of 5 -AMP-activated Protein Kinase in the Stimulation of Glucose Transport in Response
More informationUnder most conditions, glucose transport is the
Rapid Publication Metabolic Stress and Altered Glucose Tr a n s p o r t Activation of AMP-Activated Protein Kinase as a Unifying Coupling Mechanism Tatsuya Hayashi, Michael F. Hirshman, Nobuharu Fujii,
More informationWilliam G. Aschenbach, Michael F. Hirshman, Nobuharu Fujii, Kei Sakamoto, Kirsten F. Howlett, and Laurie J. Goodyear
Effect of AICAR Treatment on Glycogen Metabolism in Skeletal Muscle William G. Aschenbach, Michael F. Hirshman, Nobuharu Fujii, Kei Sakamoto, Kirsten F. Howlett, and Laurie J. Goodyear AMP-activated protein
More informationExercise is an important component of the treatment
Rapid Publication AMP-Activated Protein Kinase (AMPK) Is Activated in Muscle of Subjects With Type 2 Diabetes During Exercise Nicolas Musi, 1 Nobuharu Fujii, 1 Michael F. Hirshman, 1 Ingvar Ekberg, 2 Sven
More informationSupplemental Figure I
Supplemental Figure I Kl ( mmol/l)-induced Force orta M (mn) 1 (mn) 1 Supplemental Figure I. Kl-induced contractions. and, Kl ( mmol/l)-induced contractions of the aorta () and those of mesenteric arteries
More informationOriginal Article The activation of AMPK in cardiomyocytes at the very early stage of hypoxia relies on an adenine nucleotide-independent mechanism
Int J Clin Exp Pathol 2012;5(8):770-776 www.ijcep.com /ISSN:1936-2625/IJCEP1207018 Original Article The activation of AMPK in cardiomyocytes at the very early stage of hypoxia relies on an adenine nucleotide-independent
More informationSUPPLEMENTARY INFORMATION
Supplementary Figures Supplementary Figure S1. Binding of full-length OGT and deletion mutants to PIP strips (Echelon Biosciences). Supplementary Figure S2. Binding of the OGT (919-1036) fragments with
More information24 th November 2008 Glasgow eprints Service https://eprints.gla.ac.uk
Ewart, M-A. and Kohlhaas, C.F. and Salt, I.P. (2008) Inhibition of tumor necrosis factor α stimulated monocyte adhesion to human aortic endothelial cells by AMP-activated protein kinase. Arteriosclerosis,
More informationMetformin and phenformin are derivatives of
The Antidiabetic Drug Metformin Activates the AMP-Activated Protein Kinase Cascade via an Adenine Nucleotide-Independent Mechanism Simon A. Hawley, 1 Anne E. Gadalla, 1 Grith Skytte Olsen, 2 and D. Grahame
More informationAMPK as a metabolic switch in rat muscle, liver and adipose tissue after exercise
Acta Physiol Scand 23, 78, 43 442 AMPK as a metabolic switch in rat muscle, liver and adipose tissue after exercise N. B. Ruderman, H. Park, V. K. Kaushik, D. Dean, S. Constant, M. Prentki 2 and A. K.
More informationThe rabbit femoral artery was prepared and each arterial ring was permeabilized
Online Supplement Nakmura et al. cgmp-dependent relaxation of smooth muscle Materials and Methods Measurement of tension The rabbit femoral artery was prepared and each arterial ring was permeabilized
More informationStudies in a wide variety of cultured cells have
Glucose Autoregulates Its Uptake in Skeletal Muscle Involvement of AMP-Activated Protein Kinase Samar I. Itani, Asish K. Saha, Theodore G. Kurowski, Heather R. Coffin, Keith Tornheim, and Neil B. Ruderman
More informationInsulin Signaling After Exercise in Insulin Receptor Substrate-2 Deficient Mice
Insulin Signaling After Exercise in Insulin Receptor Substrate-2 Deficient Mice Kirsten F. Howlett, Kei Sakamoto, Michael F. Hirshman, William G. Aschenbach, Matthew Dow, Morris F. White, and Laurie J.
More informationCell Signaling part 2
15 Cell Signaling part 2 Functions of Cell Surface Receptors Other cell surface receptors are directly linked to intracellular enzymes. The largest family of these is the receptor protein tyrosine kinases,
More informationAMP-Activated Protein Kinase Conducts the Ischemic Stress Response Orchestra Lawrence H. Young. doi: /CIRCULATIONAHA.107.
AMP-Activated Protein Kinase Conducts the Ischemic Stress Response Orchestra Lawrence H. Young Circulation. 2008;117:832-840 doi: 10.1161/CIRCULATIONAHA.107.713115 Circulation is published by the American
More informationPhosphorylation-activity relationships of AMPK and acetyl-coa carboxylase in muscle
J Appl Physiol 92: 2475 2482, 2002; 10.1152/japplphysiol.00071.2002. Phosphorylation-activity relationships of AMPK and acetyl-coa carboxylase in muscle S. H. PARK, S. R. GAMMON, J. D. KNIPPERS, S. R.
More informationPRODUCT INFORMATION & MANUAL
PRODUCT INFORMATION & MANUAL Mitochondrial Extraction Kit NBP2-29448 Research use only. Not for diagnostic or therapeutic procedures www.novusbio.com P: 303.760.1950 P: 888.506.6887 F: 303.730.1966 technical@novusbio.com
More informationAMPK and p38 MAPK Participate in the Stimulation of. Glucose Uptake by Dinitrophenol in Adult Cardiomyocytes
Endocrinology. First published January 27, 2005 as doi:10.1210/en.2004-1565 AMPK and p38 MAPK Participate in the Stimulation of Glucose Uptake by Dinitrophenol in Adult Cardiomyocytes Abbreviated title:
More informationAMPK a 1 Activation Is Required for Stimulation of Glucose Uptake by Twitch Contraction, but Not by H 2 O 2, in Mouse Skeletal Muscle
AMPK a 1 Activation Is Required for Stimulation of Glucose Uptake by Twitch Contraction, but Not by H 2 O 2, in Mouse Skeletal Muscle Thomas E. Jensen 1, Peter Schjerling 2,3, Benoit Viollet 4,5, Jørgen
More informationInsulin increases glucose transport activity in muscle
5-Aminoimidazole-4-Carboxamide Ribonucleoside (AICAR) Inhibits Insulin-Stimulated Glucose Transport in 3T3-L1 Adipocytes Ian P. Salt, John M. C. Connell, and Gwyn W. Gould Incubation of skeletal muscle
More informationWestern Immunoblotting Preparation of Samples:
Western Immunoblotting Preparation of Samples: Total Protein Extraction from Culture Cells: Take off the medium Wash culture with 1 x PBS 1 ml hot Cell-lysis Solution into T75 flask Scrap out the cells
More informationChinese Bulletin of Life Sciences AMP. (AMP-activated protein kinase, AMPK) Advances in the studies of AMP-activated protein kinase
17 2 2005 4 Chinese Bulletin of Life Sciences Vol. 17, No. 2 Apr., 2005 1004-0374(2005)02-0147-06 400038 AMP (AMP-activated protein kinase, ) AMP Q555.7 A Advances in the studies of AMP-activated protein
More informationEffects and mechanisms of Fenofibrate on the secretion of vascular endothelial contraction factors in hypertensive rats
Effects and mechanisms of Fenofibrate on the secretion of vascular endothelial contraction factors in hypertensive rats Y. Zhu 1, H.-S. Wang 1, X.-M. Li 1 and C. Qu 2 1 Department of Cardiac Surgery, General
More informationAMPK Assay. Require: Sigma (1L, $18.30) A4206 Aluminum foil
AMPK Assay Require: Acetone Sigma (1L, $18.30) A4206 Aluminum foil Ammonium sulfate Fisher BP212R-1 AMP Sigma A1752 ATP Sigma A6144 (alt. use A7699) Beta-mercaptoethanol Sigma M6250 (alt. use M7154) Bio-Rad
More informationPHYSIOLOGY, ENDOCRINOLOGY, AND REPRODUCTION. Research Note. Energy sensing in developing chicken embryos and posthatch chicks from different size eggs
PHYSIOLOGY, ENDOCRINOLOGY, AND REPRODUCTION Research Note Energy sensing in developing chicken embryos and posthatch chicks from different size eggs Q. Hu, U. Agarwal, and B. J. Bequette 1 Animal and Avian
More informationEffect of AMPK activation on muscle glucose metabolism in conscious rats
Effect of AMPK activation on muscle glucose metabolism in conscious rats RAYNALD BERGERON, 1 RAYMOND R. RUSSELL III, 1 LAWRENCE H. YOUNG, 1 JIAN-MING REN, 2 MELISSA MARCUCCI, 1 AGNES LEE, 1 AND GERALD
More informationduring low-intensity exercise
Am J Physiol Endocrinol Metab 296: E47 E55, 2009. First published October 21, 2008; doi:10.1152/ajpendo.90690.2008. 2 -AMPK activity is not essential for an increase in fatty acid oxidation during low-intensity
More informationModulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis
icell Skeletal Myoblasts Application Protocol Modulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis Introduction The skeletal muscle is one of the
More informationEffects of sitagliptin on cardiac metabolism in mice
Effects of sitagliptin on cardiac metabolism in mice M. Lenski, J.-C. Reil, M. Böhm, U. Laufs Saarland University Hospital Department of Internal Medicine III, Cardiology Homburg - Germany Disclosures
More informationNitric oxide stimulates glucose transport through insulin-independent GLUT4 translocation in 3T3-L1 adipocytes
European Journal of Endocrinology (2003) 149 61 67 ISSN 0804-4643 EXPERIMENTAL STUDY Nitric oxide stimulates glucose transport through insulin-independent GLUT4 translocation in 3T3-L1 adipocytes Takashi
More informationWe thank Dr. Bell for his interest in our paper. He states that metformin has never been
ENDOCRINE PRACTICE Rapid Electronic Article in Press Rapid Electronic Articles in Press are preprinted manuscripts that have been reviewed and accepted for publication, but have yet to be edited, typeset
More informationNOS Activity Assay Kit
NOS Activity Assay Kit Catalog Number KA1345 50 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Principle of the Assay... 3 General Information... 4
More informationExercise in Adverse Cardiac Remodeling: of Mice and Men
Exercise in Adverse Cardiac Remodeling: of Mice and Men 17-01-2013 Dirk J Duncker Experimental Cardiology, Cardiology, Thoraxcenter Cardiovascular Research Institute COEUR Erasmus MC, University Medical
More informationPKA as an Upstream Kinase for LKB1/STRAD/ MO25
Brigham Young University BYU ScholarsArchive All Theses and Dissertations 2006-07-10 PKA as an Upstream Kinase for LKB1/STRAD/ MO25 Seth Taylor Herway Brigham Young University - Provo Follow this and additional
More information2013 W. H. Freeman and Company. 12 Signal Transduction
2013 W. H. Freeman and Company 12 Signal Transduction CHAPTER 12 Signal Transduction Key topics: General features of signal transduction Structure and function of G protein coupled receptors Structure
More informationAMPK activity and isoform protein expression are similar in muscle of obese subjects with and without type 2 diabetes
Am J Physiol Endocrinol Metab 286: E239 E244, 2004. First published October 7, 2003; 10.1152/ajpendo.00326.2003. AMPK activity and isoform protein expression are similar in muscle of obese subjects with
More informationGrowth and Differentiation Phosphorylation Sampler Kit
Growth and Differentiation Phosphorylation Sampler Kit E 0 5 1 0 1 4 Kits Includes Cat. Quantity Application Reactivity Source Akt (Phospho-Ser473) E011054-1 50μg/50μl IHC, WB Human, Mouse, Rat Rabbit
More informationGlycogen synthase (GS) catalyzes a crucial and
Regulation of Glycogen Synthase by Glucose and Glycogen A Possible Role for AMP-Activated Protein Kinase Reza Halse, 1 Lee G.D. Fryer, 2 James G. McCormack, 3 David Carling, 2 and Stephen J. Yeaman 1 We
More informationPhospho-AKT Sampler Kit
Phospho-AKT Sampler Kit E 0 5 1 0 0 3 Kits Includes Cat. Quantity Application Reactivity Source Akt (Ab-473) Antibody E021054-1 50μg/50μl IHC, WB Human, Mouse, Rat Rabbit Akt (Phospho-Ser473) Antibody
More informationAMPK Phosphorylation Assay Kit
AMPK Phosphorylation Assay Kit Catalog Number KA3789 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 Principle
More informationFuel the Failing Heart: glucose or fatty acids? Rong Tian, MD, PhD Mitochondria and Metabolism Center University of Washington, Seattle
Fuel the Failing Heart: glucose or fatty acids? Rong Tian, MD, PhD Mitochondria and Metabolism Center University of Washington, Seattle Metabolic Remodeling: Fatty Acids Carbohydrates PCr/ATP Glucose vs.
More informationWolff-Parkinson-White Syndrome and PRKAG2
Wolff-Parkinson-White Syndrome and PRKAG2 Maggie Beatka University of Wisconsin-Madison http://www.beatmap.net/portfolio-detail/human-cardiovascular-system-3drenderings/ What causes Wolff-Parkinson-White?
More informationBALANCING THE SCALES USING A NOVEL CELLULAR ENERGY SENSOR
The West London Medical Journal 2010 Vol 2 No 4 pp 29-35 BALANCING THE SCALES USING A NOVEL CELLULAR ENERGY SENSOR Sairah Akbar The topic of obesity is rarely out of the public eye with an increasingly
More informationAMPK is a major regulator of cellular and wholebody
ORIGINAL ARTICLE Molecular Mechanism by Which AMP-Activated Protein Kinase Activation Promotes Glycogen Accumulation in Muscle Roger W. Hunter, 1 Jonas T. Treebak, 2 Jørgen F.P. Wojtaszewski, 2 and Kei
More informationReactive oxygen species: Importance for ischemia/reperfusion (injury)
Physiologisches Institut Reactive oxygen species: Importance for ischemia/reperfusion (injury) Prof. Dr. Rainer Schulz Reactive oxygen species (ROS) in ischemia/reperfusion injury (IRI) ROS GOOD: Endogenous
More informationMODULATION OF AMP DEAMINASE IN RAT HEARTS SUBJECTED TO ISCHEMIA AND REPERFUSION BY PURINE RIBOSIDE
Nucleosides, Nucleotides, and Nucleic Acids, 27:876 88, 28 Copyright C Taylor & Francis Group, LLC ISSN: 1525-777 print / 1532-2335 online DOI: 1.18/152577782146551 MODULATION OF AMP DEAMINASE IN RAT HEARTS
More informationRevision. General functions of hormones. Hormone receptors. Hormone derived from steroids Small polypeptide Hormone
االله الرحمن الرحيم بسم Revision General functions of hormones. Hormone receptors Classification according to chemical nature Classification according to mechanism of action Compare and contrast between
More informationSUPPLEMENTAL MATERIAL. Supplementary Methods
SUPPLEMENTAL MATERIAL Supplementary Methods Culture of cardiomyocytes, fibroblasts and cardiac microvascular endothelial cells The isolation and culturing of neonatal rat ventricular cardiomyocytes was
More informationAMP kinase expression and activity in human skeletal muscle: effects of immobilization, retraining, and creatine supplementation
J Appl Physiol 98: 1228 1233, 2005. First published October 29, 2004; doi:10.1152/japplphysiol.00665.2004. AMP kinase expression and activity in human skeletal muscle: effects of immobilization, retraining,
More informationBBSG 501 Section 4 Metabolic Fuels, Energy and Order Fall 2003 Semester
BBSG 501 Section 4 Metabolic Fuels, Energy and Order Fall 2003 Semester Section Director: Dave Ford, Ph.D. Office: MS 141: ext. 8129: e-mail: fordda@slu.edu Lecturers: Michael Moxley, Ph.D. Office: MS
More informationResearch Paper Correspondence: Louis Hue A-S.M. and L.B. contributed equally to this work.
Research Paper 1247 Phosphorylation and activation of heart PFK-2 by has a role in the stimulation of glycolysis during ischaemia A-S. Marsin, L. Bertrand, M.H. Rider, J. Deprez, C. Beauloye, M.F. Vincent,
More informationCellular Physiology (PHSI3009) Contents:
Cellular Physiology (PHSI3009) Contents: Cell membranes and communication 2 nd messenger systems G-coupled protein signalling Calcium signalling Small G-protein signalling o RAS o MAPK o PI3K RHO GTPases
More informationProtocol for Gene Transfection & Western Blotting
The schedule and the manual of basic techniques for cell culture Advanced Protocol for Gene Transfection & Western Blotting Schedule Day 1 26/07/2008 Transfection Day 3 28/07/2008 Cell lysis Immunoprecipitation
More informationCauses Acute Hepatic Insulin Resistance In Vivo
5-Aminoimidazole-4-Carboxamide-1- -D-Ribofuranoside Causes Acute Hepatic Insulin Resistance In Vivo R. Richard Pencek, Jane Shearer, Raul C. Camacho, Freyja D. James, D. Brooks Lacy, Patrick T. Fueger,
More informationIn vitro functional study of Cardiomyocyte
In vitro functional study of Cardiomyocyte Gwang Hyeon Eom Department of Pharmacology and Medical Research Center for Gene Regulation Chonnam National University Medical School, Gwangju, South Korea Heart
More informationThe 5 AMP-activated protein kinase (AMPK) is a
Original Article AMPK-Mediated AS160 Phosphorylation in Skeletal Muscle Is Dependent on AMPK Catalytic and Regulatory Subunits Jonas T. Treebak, 1 Stephan Glund, 2 Atul Deshmukh, 2 Ditte K. Klein, 1 Yun
More information7/31/2009. G.Y. Prince Used Cars 10 am Los Angelos, CA Mullholland Drive..later that day. Would you buy a car without taking it for a spin first?
7/31/29 My Anna will love it! Who needs a test drive? Or a Warranty? It looked great in the lot! Do mean to say that you never actually test drove the car? G.Y. Prince Used Cars 1 am Los Angelos, CA Mullholland
More informationASSAY OF SPHINGOMYELINASE ACTIVITY
ASSAY OF SPHINGOMYELINASE ACTIVITY Protocol for Protein Extraction Stock Solution 1. Leupeptin/hydrochloride (FW 463.0,
More informationExperimental Physiology
Exp Physiol 99.12 (2014) pp 1569 1573 1569 Symposium Report Symposium Report Role of nitric oxide in skeletal muscle glucose uptake during exercise Yet Hoi Hong 1,2,3, Andrew C. Betik 1,2 and Glenn K.
More informationEffec<ve Use of PI3K and MEK Inhibitors to Treat Mutant K Ras G12D and PIK3CA H1047R Murine Lung Cancers
Effec
More informationExercise is a fundamental aspect of type 2 diabetes
Original Article Effect of Acute Exercise on AMPK Signaling in Skeletal Muscle of Subjects With Type 2 Diabetes A Time-Course and Dose-Response Study Apiradee Sriwijitkamol, 1,2 Dawn K. Coletta, 1 Estela
More informationElectronic supplementary material (ESM) MATERIALS AND METHODS. Study subjects.
Electronic supplementary material (ESM) MATERIALS AND METHODS Study subjects. Twelve obese patients with type 2 diabetes carefully matched to ten healthy, lean and ten obese, non-diabetic volunteers participated
More informationMetformin is one of the most commonly prescribed
ORIGINAL ARTICLE Acute Metformin Therapy Confers Cardioprotection Against Myocardial Infarction Via AMPK-eNOS Mediated Signaling John W. Calvert, 1 Susheel Gundewar, 1 Saurabh Jha, 1 James J.M. Greer,
More informationThe Schedule and the Manual of Basic Techniques for Cell Culture
The Schedule and the Manual of Basic Techniques for Cell Culture 1 Materials Calcium Phosphate Transfection Kit: Invitrogen Cat.No.K2780-01 Falcon tube (Cat No.35-2054:12 x 75 mm, 5 ml tube) Cell: 293
More informationAcute Metformin Therapy Confers Cardioprotection Against Myocardial Infarction Via AMPK-eNOS Mediated Signaling
Diabetes Publish Ahead of Print, published online December 14, 2007 Acute Metformin Therapy Confers Cardioprotection Against Myocardial Infarction Via AMPK-eNOS Mediated Signaling John W. Calvert 1, Ph.D.,
More informationStroke is the third leading cause of death behind heart
Neuroprotective Effects of Adenosine Monophosphate- Activated Protein Kinase Inhibition and Gene Deletion in Stroke Jun Li, PhD; Zhiyuan Zeng, BS; Benoit Viollet, PhD; Gabriele V. Ronnett, MD, PhD; Louise
More informationGENERAL CHARACTERISTICS OF THE ENDOCRINE SYSTEM FIGURE 17.1
GENERAL CHARACTERISTICS OF THE ENDOCRINE SYSTEM FIGURE 17.1 1. The endocrine system consists of glands that secrete chemical signals, called hormones, into the blood. In addition, other organs and cells
More informationExercise modulates postreceptor insulin signaling and glucose transport in muscle-specific insulin receptor knockout mice
Exercise modulates postreceptor insulin signaling and glucose transport in muscle-specific insulin receptor knockout mice Jørgen F.P. Wojtaszewski, Yasuki Higaki, Michael F. Hirshman, M. Dodson Michael,
More informationTHE GLUCOSE-FATTY ACID-KETONE BODY CYCLE Role of ketone bodies as respiratory substrates and metabolic signals
Br. J. Anaesth. (1981), 53, 131 THE GLUCOSE-FATTY ACID-KETONE BODY CYCLE Role of ketone bodies as respiratory substrates and metabolic signals J. C. STANLEY In this paper, the glucose-fatty acid cycle
More informationFEBS 1138 January Paul R. Buckland and Bernard Rees Smith
Volume 166, number 1 FEBS 1138 January 1984 A structural comparison receptors by of guinea pig thyroid and fat TSH photoaffinity labelling Paul R. Buckland and Bernard Rees Smith Endocrine Immunology Unit,
More informationMOLECULAR AND CELLULAR BIOLOGY, Sept. 2000, p Vol. 20, No. 18. Copyright 2000, American Society for Microbiology. All Rights Reserved.
MOLECULAR AND CELLULAR BIOLOGY, Sept. 2000, p. 6704 6711 Vol. 20, No. 18 0270-7306/00/$04.00 0 Copyright 2000, American Society for Microbiology. All Rights Reserved. Characterization of the Role of AMP-Activated
More informationExercise Effects of Muscle Insulin Signaling and Action Invited Review: Exercise training-induced changes in insulin signaling in skeletal muscle
J Appl Physiol 93: 773 781, 2002; 10.1152/japplphysiol.00126.2002. highlighted topics Exercise Effects of Muscle Insulin Signaling and Action Invited Review: Exercise training-induced changes in insulin
More informationREGULATION OF SKELETAL MUSCLE GLUCOSE UPTAKE DURING CONTRACTION/ EXERCISE BY NITRIC OXIDE (NO)/ NEURONAL NITRIC OXIDE SYNTHASE MU (nnosμ)
REGULATION OF SKELETAL MUSCLE GLUCOSE UPTAKE DURING CONTRACTION/ EXERCISE BY NITRIC OXIDE (NO)/ NEURONAL NITRIC OXIDE SYNTHASE MU (nnosμ) YET HOI HONG (MBBS, MMedSc) Submitted in fulfillment of the requirements
More informationPrior AICAR Stimulation Increases Insulin Sensitivity in Mouse Skeletal Muscle in an AMPK-Dependent Manner
2042 Diabetes Volume 64, June 2015 Rasmus Kjøbsted, 1,2 Jonas T. Treebak, 1,2 Joachim Fentz, 1 Louise Lantier, 3,4,5 Benoit Viollet, 3,4,5 Jesper B. Birk, 1 Peter Schjerling, 6 Marie Björnholm, 7 Juleen
More information- Biosignaling: Signal transduction. References: chapter 8 of Lippincots chapter 1 3 of Lehningers
Basic concepts of Metabolism Metabolism and metabolic pathway Metabolic Map Catabolism Anabolism - Regulation of Metabolism Signals from within the cell (Intracellular) Communication between cells. - Biosignaling:
More informationAPPENDIX Heparin 2 mg heparin was dissolved in 0.9 % NaCl (10 ml). 200 µl of heparin was added to each 1 ml of blood to prevent coagulation.
APPENDIX 1 Preparation of reagents 1.1. Preparation of dosing solution Nonylphenol 15 mg of Nonylphenol was dissolved in olive oil (10 ml) and used as stock solution. The stock solution was serially diluted
More informationChromatin IP (Isw2) Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles.
Chromatin IP (Isw2) 7/01 Toshi last update: 06/15 Reagents Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles. 2.5 M glycine. TBS:
More information,, - [5, 11]., -, (NO)., NO,,. NO NO- (NOS). (nnos NOS1) (enos NOS3) NO-,, 2+- NO- (inos NOS2),,,,, [5]., NOS (nnos enos), inos. inos - ( ),, ( ),. NO
, 70,9 ± 1,3% 58,0±0,86%. 3... 1.. /..,.. // today. 2004. 1-2.. 30-31. 2.. /.,..,.. //-.: -, -2006. - 240. 3... /.., C. B.,.. [.] //. -2008. - 5.-C.33-36. 4..., - /..,.. // - 2007. - 1. -. 38-41. 5...
More informationMetformin inhibits hepatic gluconeogenesis in mice independently of the LKB1/AMPK pathway via a decrease in hepatic energy state
Related Commentary, page 2267 Research article Metformin inhibits hepatic gluconeogenesis in mice independently of the LKB1/AMPK pathway via a decrease in hepatic energy state Marc Foretz, 1,2 Sophie Hébrard,
More informationREGULATION OF THE GLUCOSE TRANSPORT PATHWAY IN ADIPOSE TISSUE OF HORSES WITH INSULIN RESISTANCE
REGULATION OF THE GLUCOSE TRANSPORT PATHWAY IN ADIPOSE TISSUE OF HORSES WITH INSULIN RESISTANCE Undergraduate Thesis as Required for Graduation with Distinction Kaleb Kohler May 23, 2011 Research Advisor:
More informationRelationship between apoptosis and alteration of the energetic metabolism pathways of hypertrophic cardiomyocytes induced by hypoxia-reoxygenation
636 Acta Physiologica Sinica, October 25, 2005, 57 (5): 636-642 http://www.actaps.com.cn * 400037 0.1 µmol/l 1 µmol/l 95% N 2 5% CO 2 5 mmhg 8 h (pyruvate dehydrogenase, PDH) -1(carnitine palmitoyltransferase
More informationELISA Kit Catalog #KHO0651
ELISA Kit Catalog #KHO0651 AMPKα [pt172]* www.invitrogen.com Invitrogen Corporation 542 Flynn Road, Camarillo, CA 93012 Tel: 800-955-6288 E-mail: techsupport@invitrogen.com *Patent Pending 1 2 TABLE OF
More informationGinkgo biloba extract postconditioning reduces myocardial ischemia reperfusion injury
Ginkgo biloba extract postconditioning reduces myocardial ischemia reperfusion injury K. Ran 1, D.-L. Yang 1, Y.-T. Chang 1, K.-M. Duan 2, Y.-W. Ou 2, H.-P. Wang 3 and Z.-J. Li 1 1 Department of Anesthesiology,
More informationAMP-Activated Protein Kinase Activation Preconditions the Heart against Ischemic Injury
Yale University EliScholar A Digital Platform for Scholarly Publishing at Yale Yale Medicine Thesis Digital Library School of Medicine 11-9-2009 AMP-Activated Protein Kinase Activation Preconditions the
More information