Acetylation has recently emerged as an important mechanism for controlling a broad array of proteins mediating cellular adaptation to metabolic fuels. Acetylation is governed, in part, by SIRTs (sirtuins), class III NAD+-dependent deacetylases that regulate lipid and glucose metabolism in liver during fasting and aging. However, the role of acetylation or SIRTs in pathogenic hepatic fuel metabolism under nutrient excess is unknown. In the present study, we isolated acetylated proteins from total liver proteome and observed 193 preferentially acetylated proteins in mice fed on an HFD (high-fat diet) compared with controls, including 11 proteins not previously identified in acetylation studies. Exposure to the HFD led to hyperacetylation of proteins involved in gluconeogenesis, mitochondrial oxidative metabolism, methionine metabolism, liver injury and the ER (endoplasmic reticulum) stress response. Livers of mice fed on the HFD had reduced SIRT3 activity, a 3-fold decrease in hepatic NAD+ levels and increased mitochondrial protein oxidation. In contrast, neither SIRT1 nor histone acetyltransferase activities were altered, implicating SIRT3 as a dominant factor contributing to the observed phenotype. In Sirt3−/− mice, exposure to the HFD further increased the acetylation status of liver proteins and reduced the activity of respiratory complexes III and IV. This is the first study to identify acetylation patterns in liver proteins of HFD-fed mice. Our results suggest that SIRT3 is an integral regulator of mitochondrial function and its depletion results in hyperacetylation of critical mitochondrial proteins that protect against hepatic lipotoxicity under conditions of nutrient excess.

Cells are exposed during their lifetimes to an array of physical forces ranging from those generated by association with other cells and extracellular matrices to the constant forces placed on cells by gravity. Alterations in these forces, either with differentiation and development or changes in activity or behavior, result in modifications in the biochemistry and adaptation in structure and function of cells. Also, a variety of differentiated cells have unique shapes that relate to extremely specialized functions, with structure and function emerging concurrently. These observations lead to the concept that the forces perceived by cells may dictate their shape, and the combined effects of external physical stimuli and internal forces responsible for maintaining cell shape may stimulate alterations in cellular biochemistry. This review examines the state of our knowledge concerning the mechanisms through which physical forces are converted to biochemical signals (mechanotransduction), and speculates on the molecular structures that may be involved in mechanotransduction.

Objective: This study was performed to assess the utility of selective small-molecule inhibitors of class I HDACs in a preclinical model of pulmonary hypertension. Methods and Results:Rats were exposed to hypobaric hypoxia for 3 weeks in the absence or presence of a benzamide HDAC inhibitor, MGCD0103, which selectively inhibits class I HDACs 1, 2, and 3. The compound reduced pulmonary arterial pressure more dramatically than tadalafil, a standard-of-care therapy for human pulmonary hypertension that functions as a vasodilator. MGCD0103 improved pulmonary artery acceleration time and reduced systolic notching of the pulmonary artery flow envelope, which suggests a positive impact of the HDAC inhibitor on pulmonary vascular remodeling and stiffening. Similar results were obtained with an independent class I HDAC-selective inhibitor, MS-275. Reduced pulmonary arterial pressure in MGCD0103-treated animals was associated with blunted pulmonary arterial wall thickening because of suppression of smooth muscle cell proliferation. Right ventricular function was maintained in MGCD0103-treated animals. Although the class I HDAC inhibitor only modestly reduced right ventricular hypertrophy, it had multiple beneficial effects on the right ventricle, which included suppression of pathological gene expression, inhibition of proapoptotic caspase activity, and repression of proinflammatory protein expression. Key Words: histone deacetylase Ⅲ pulmonary hypertension Ⅲ proliferation Ⅲ gene expression Ⅲ signaling pathways I n patients with pulmonary hypertension (PH), restricted blood flow through the pulmonary arterial circulation due to increased pulmonary vascular resistance often results in right-sided heart failure. Despite recent advances in the treatment of PH, the 5-year mortality rate for individuals with this disease still approaches 50%, which highlights an urgent need for novel therapeutics. 1 Current standards of care for patients with PH typically involve the use of vasoactive drugs, including endothelin receptor antagonists, phosphodiesterase type 5 inhibitors, and prostacyclins. More effective therapeutic strategies will likely be based on the combined use of vasodilators and agents that target distinct pathogenic mechanisms in PH, such as pulmonary vascular inflammation and fibrosis, as well as aberrant proliferation of smooth muscle cells, endothelial cells, and fibroblasts in the lung vasculature. 2 Additionally, because right ventricular (RV) failure is the cause of death in the majority of PH patients, 3,4 and it is unclear whether standards of care for left ventricular Original received October 8, 2011; revision received January 11, 2012; accepted January 18, 2012. In December 2011 HDACs control cell proliferation, inflammation, and fibrosis by catalyzing removal of acetyl groups from lysine residues in a variety of proteins. The 18 mammalian HDACs are encoded by distinct genes and are grouped into 4 classes. 6 Two broad-spectrum HDAC inhibitors are approved for the treatment of cancer. One of these compounds,...

This study was conducted to examine the role of myocardial ATP-sensitive potassium (K ATP ) channels in exercise-induced protection from ischaemia-reperfusion (I-R) injury. Female rats were either sedentary (Sed) or exercised for 12 weeks (Tr). Hearts were excised and underwent a 1-2 h regional I-R protocol. Prior to ischaemia, hearts were subjected to pharmacological blockade of the sarcolemmal K ATP channel with HMR 1098 (SedHMR and TrHMR), mitochondrial blockade with 5-hydroxydecanoic acid (5HD; Sed5HD and Tr5HD), or perfused with buffer containing no drug (Sed and Tr). Infarct size was significantly smaller in hearts from Tr animals (35.4 ± 2.3 versus 44.7 ± 3.0% of the zone at risk for Tr and Sed, respectively). Mitochondrial K ATP blockade did not abolish the training-induced infarct size reduction (30.0 ± 3.4 versus 38.0 ± 2.6 in Tr5HD and Sed5HD, respectively); however, sarcolemmal K ATP blockade completely eradicated the training-induced cardioprotection. Infarct size was 71.2 ± 3.3 and 64.0 ± 2.4% of the zone at risk for TrHMR and Sed HMR. The role of sarcolemmal K ATP channels in Tr-induced protection was also supported by significant increases in both subunits of the sarcolemmal K ATP channel following training. LV developed pressure was better preserved in hearts from Tr animals, and was not influenced by addition of HMR 1098. 5HD decreased pressure development regardless of training status, from 15 min of ischaemia through the duration of the protocol. This mechanical dysfunction was likely to be due to a 5HD-induced increase in myocardial Ca 2+ content following I-R. The major findings of the present study are: (1) unlike all other known forms of delayed cardioprotection, infarct sparing following chronic exercise was not abolished by 5HD; (2) pharmacological blockade of the sarcolemmal K ATP channel nullified the cardioprotective benefits of exercise training; and (3) increased expression of sarcolemmal K ATP channels was observed following chronic training.

Abstract-Hypertrophic cardiomyopathy (HCM) is the most common form of sudden death in young competitive athletes.However, exercise has also been shown to be beneficial in the setting of other cardiac diseases. We examined the ability of voluntary exercise to prevent or reverse the phenotypes of a murine model of HCM harboring a mutant myosin heavy chain (MyHC). No differences in voluntary cage wheel performance between nontransgenic (NTG) and HCM male mice were seen. Exercise prevented fibrosis, myocyte disarray, and induction of "hypertrophic" markers including NFAT activity when initiated before established HCM pathology. If initiated in older HCM animals with documented disease, exercise reversed myocyte disarray (but not fibrosis) and "hypertrophic" marker induction. In addition, exercise returned the increased levels of phosphorylated GSK-3␤ to those of NTG and decreased levels of phosphorylated CREB in HCM mice to normal levels. Exercise in HCM mice also favorably impacted components of the apoptotic signaling pathway, including Bcl-2 (an inhibitor of apoptosis) and procaspase-9 (an effector of apoptosis) expression, and caspase-3 activity. Remarkably, there were no differences in mortality between exercised NTG and HCM mice. Thus, not only was exercise not harmful but also it was able to prevent and even reverse established cardiac disease phenotypes in this HCM model. (Circ Res. 2006;98:540-548.)Key Words: apoptosis Ⅲ exercise Ⅲ hypertrophic cardiomyopathy Ⅲ remodeling T reatment strategies for hypertrophic cardiomyopathy (HCM) depend on clinical symptoms and stratification of sudden death risk with little evidence supporting prophylactic treatment of asymptomatic HCM. 1 It is becoming increasingly evident that cardiac rehabilitation for patients with other forms of cardiac disease such as ischemic heart disease, hypertension, and congestive heart failure (CHF) includes regular exercise, particularly aerobic exercise, and that exercise reduces cardiovascular morbidity and mortality in these subjects. 2,3 Yet there are no reports in the literature that discuss the impact of mild, monitored exercise as a therapeutic measure for HCM patients. Exercise rehabilitation in HCM patients, however, is somewhat counterintuitive considering the high incidence of sudden cardiac death in young athletes and that both the American Heart Association and the European Society of Cardiology strongly encourage preparticipation cardiovascular screening of HCM for young competitive athletes. 4,5 However, we hypothesized that voluntary cage wheel running as a form of mild exercise would improve and/or prevent the pathologic cardiac phenotype in a murine model of HCM. Voluntary cage wheel was chosen as the exercise intervention because it eliminates physical and psychological stressors associated with forced exercise paradigms that may exacerbate the HCM syndrome, and we have previously demonstrated cardiac adaptation to voluntary cage wheel exercise. 6 The particular HCM mouse model developed by our group 7 is ideal to addres...

We hypothesized that cAMP response element-binding protein (CREB) could function as a molecular determinant of smooth muscle cell fate. In arterial sections from the systemic and pulmonary circulation, CREB content was high in proliferation-resistant medial subpopulations of smooth muscle cells and low in proliferation-prone regions. In vessels from neonatal calves exposed to chronic hypoxia, CREB content was depleted and smooth muscle cell (SMC) proliferation was accelerated. Induction of quiescence by serum deprivation in culture led to increased CREB content. Highly proliferative SMC in culture were observed to have low CREB content. Exposure to proliferative stimuli such as hypoxia or platelet-derived growth factor decreased SMC CREB content. Assessment of CREB gene transcription by nuclear run-on analysis and transcription from a CREB promoter-luciferase construct indicate that CREB levels in SMC are in part controlled at the level of transcription. Overexpression of wild type or constitutively active CREB in primary cultures of SMC arrested cell cycle progression. Additionally, expression of constitutively active CREB decreased both proliferation and chemokinesis. Consistent with these functional properties, active CREB decreased the expression of multiple cell cycle regulatory genes, as well as genes encoding growth factors, growth factor receptors, and cytokines. Our data suggest a unique mode of cellular phenotype determination at the level of the nuclear content of CREB.The vessel wall, once seen as simply a mechanical conduit for blood flow, is a complex organ whose dysfunction is responsible for the leading cause of death in the United States, cardiovascular disease. The lumen of blood vessels is lined with endothelial cells, which communicate with the underlying medial cell layer. For many years, the arterial media was considered to be made-up of a homogeneous population of smooth muscle cells (SMC) 1 arising from a common lineage. Thus the diverse activities of contraction, proliferation, migration, and extracellular matrix production were thought to reflect SMC response to either normal or pathological stimuli. Vascular remodeling is the compensatory response of the vasculature to stress or injury. Under pathological circumstances (hypoxia, mechanical injury, hyperlipidemia, and oxidative stress), SMC in the intimal and medial compartments of the arterial wall become proliferative, migratory, and produce excess matrix proteins. This switch in SMC phenotype is termed phenotypic modulation and is considered to play a key pathogenic role in atherosclerosis and pulmonary hypertension. While the stimuli for SMC activation are well known, the molecular events, which permit activation of SMC, remain poorly understood.Cyclic nucleotides (cAMP and cGMP) promote SMC quiescence in vitro and in vivo. ␤-Adrenergic stimulation of cAMP signaling is important for SMC quiescence and contractile function under normal conditions. It is believed that cAMP acts as a gate to prevent SMC mitogenic response to growth fac...

The stratospheric polar vortex is weaker in the easterly phase of the quasi-biennial oscillation (QBO-E) than in the westerly phase (QBO-W), but the mechanism behind the QBO's influence is not well understood. The composite difference of the atmospheric state between QBO-E and QBO-W is found to closely resemble the structure of the northern annular mode, the leading empirical orthogonal function of stratospheric variability, including its wave components. Studies of dynamical systems indicate that many different forcings could give rise to this response, and therefore this composite difference does not provide much information about the forcing mechanism. It is argued that the full transient response of a system to an applied forcing is likely to be much more informative about the dynamics of the forcing mechanism, especially the response on time scales shorter than the dynamical time scale, which is about a week for vortex variability. It is shown that the transient response of the vortex to forcing by the QBO in a general circulation model is consistent with the proposed mechanism of Holton and Tan, indicating that this mechanism has a role in the QBO modulation of vortex strength, in contrast to the conclusions of several recent studies. This novel approach of examining the transient response to a forcing on short time scales may be useful in various other outstanding problems.

We hypothesized that diabetes and glucose-induced reactive oxygen species lead to depletion of cAMP response element-binding protein (CREB) content in the vasculature. In primary cultures of smooth muscle cells (SMC) high medium glucose decreased CREB function but increased SMC chemokinesis and entry into the cell cycle. These effects were blocked by pretreatment with the antioxidants. High glucose increased intracellular reactive oxygen species detected by CM-H 2 DCFA. SMC exposed to oxidative stress (H 2 O 2 ) demonstrated a 3.5-fold increase in chemokinesis (p < 0.05) and accelerated entry into cell cycle, accompanied by a significant decrease in CREB content. Chronic oxidative challenge similar to the microenvironment in diabetes (glucose oxidase treatment) decreases CREB content (40 -50%). Adenoviral-mediated expression of constitutively active CREB abolished the increase in chemokinesis and cell cycle progression induced by either high glucose or oxidative stress. Analysis of vessels from insulin resistant or diabetic animals indicates that CREB content is decreased in the vascular stroma. Treatment of insulinresistant animals with the insulin sensitizer rosiglitazone restores vessel wall CREB content toward that observed in normal animals. In summary, high glucose and oxidative stress decrease SMC CREB content increase chemokinesis and entry into the cell cycle, which is blocked by antioxidants or restoration of CREB content. Thus, decreased vascular CREB content could be one of the molecular mechanisms leading to increased atherosclerosis in diabetes.