Voluntary exercise attenuates obesity-associated inflammation through ghrelin expressed in macrophages.

Chronic low-level inflammation is associated with obesity and a sedentary lifestyle, causing metabolic disturbances such as insulin resistance. Exercise training has been shown to decrease chronic low-level systemic inflammation in high-fat diet (HFD)-induced obesity. However, the molecular mechanisms mediating its beneficial effects are not fully understood. Ghrelin is a peptide hormone predominantly produced in the stomach that stimulates appetite and induces growth hormone release. In addition to these well-known functions, recent studies suggest that ghrelin localizes to immune cells and exerts an anti-inflammatory effect. The purpose of the current study was to investigate the role of ghrelin expressed in macrophages in the anti-inflammatory effects of voluntary exercise training. Expression of tumor necrosis factor-α (TNF-α), monocyte chemotactic protein (MCP)-1 and F4/80 was increased in adipose tissue from mice fed a HFD (HFD mice) compared with mice fed a standard diet (SD mice), whereas the expression of these inflammatory cytokines was markedly decreased in mice performing voluntary wheel running during the feeding of a HFD (HFEx mice). The expression of TNF-α was also increased in peritoneal macrophages by a HFD and exercise training inhibited the increase of TNF-α expression. Interestingly, expression of ghrelin in peritoneal macrophages was decreased by a HFD and recovered by exercise training. Suppression of ghrelin expression by siRNA increased TNF-α expression and LPS-stimulated NF-κB activation in RAW264 cells, which is a macrophage cell line. TNF-α expression by stimulation with LPS was significantly suppressed in RAW264 cells cultured in the presence of ghrelin. These results suggest that ghrelin exerts potent anti-inflammatory effects in macrophages and functions as a mediator of the beneficial effects of exercise training.

Effect of exercise training on adipocyte-size-dependent expression of leptin and adiponectin.

AIMS: Our aim was to evaluate the effect of exercise training (TR) on adipocyte-size-dependent expression of leptin and adiponectin. MAIN METHODS: Male Wistar rats were divided into 2 groups, sedentary control (CR) and TR group, and both monitored for 9weeks. Adipocytes isolated from epididymal, retroperitoneal, and inguinal fat depots were independently separated into 3 fractions of different cell size, and the relationships between adipocyte size and either leptin or adiponectin mRNA were determined by real-time RT-PCR analysis. KEY FINDINGS: In epididymal and inguinal adipose tissue, positive relationships between adipocyte size and both leptin and adiponectin mRNA expression were found. Comparison of TR and CR rats showed no significant effect of TR on the slopes of the linear regression lines of correlation between leptin mRNA and adipocyte size in either adipose tissue, whereas the slopes of the regression line of correlation between adipocyte size and adiponectin mRNA were greater in TR group. Leptin levels per milliliter of plasma were significantly lower in TR than CR rats, whereas leptin levels adjusted to the 3 fat depots did not differ. TR did not affect adiponectin levels in plasma, whereas adiponectin levels adjusted to the 3 fat depots were significantly greater in TR than CR group. SIGNIFICANCE: TR-induced reduction in leptin mRNA expression was closely associated with smaller adipocyte size. However, TR amplified the adipocyte-size-dependent expression of adiponectin mRNA, suggesting that TR-induced alterations in adiponectin mRNA may also be mediated by factor(s) other than adipocyte size.

Hypoxia-inducible factor-1alpha suppresses the expression of macrophage scavenger receptor 1.

Macrophages are distributed in all peripheral tissues and play a critical role in the first line of the innate immune defenses against bacterial infection by phagocytosis of bacterial pathogens through the macrophage scavenger receptor 1 (MSR1). Within tissues, the partial pressure of oxygen (pO2) decreases depending on the distance of cells from the closest O2-supplying blood vessel. However, it is not clear how the expression of MSR1 in macrophages is regulated by low pO2. On the other hand, hypoxia-inducible factor (HIF)-1alpha is well known to control hypoxic responses through regulation of hypoxia-inducible genes. Therefore, we investigated the effects of hypoxia and HIF-1alpha on MSR1 expression and function in the macrophage cell line RAW264. Exposure to 1% O2 or treatment with the hypoxia-mimetic agent cobalt chloride (CoCl2) significantly suppressed the expression of MSR1 mRNA, accompanied by a markedly increase in levels of nuclear HIF-1alpha protein. The overexpression of HIF-1alpha in RAW264 cells suppressed the expression of MSR1 mRNA and protein, transcriptional activity of the MSR1 gene, and phagocytic capacity against the Gram-positive bacteria Listeria monocytogenes. The suppression of MSR1 mRNA by hypoxia or CoCl2 was inhibited by YC-1, an inhibitor of HIF-1alpha, or by the depletion of HIF-1alpha expression by small interference RNA. These results indicate that hypoxia transcriptionally suppresses MSR1 expression through HIF-1alpha.

Exercise training decreases expression of inflammation-related adipokines through reduction of oxidative stress in rat white adipose tissue.

Increased oxidative stress in adipocytes causes dysregulated expression of inflammation-related adipokines. We have examined the effects of exercise training on oxidative stress in rat white adipose tissue (WAT), especially focusing on inflammation-related adipokines. The levels of lipid peroxidation in WAT of exercise-trained (TR) rats were lower than those in control (C) rats. The content of manganese-containing superoxide dismutase in WAT of TR rats was increased as compared with those in C rats. In contrast, the expression of the NADPH oxidase NOX2 protein in WAT was downregulated by exercise training. Moreover, the levels of inflammation-related adipokines, such as tumor necrosis factor-alpha and monocyte chemoattractant protein-1, in WAT of TR rats were lower than those in C rats. The effects of exercise training were more remarkable in visceral WAT than in subcutaneous. These results suggest that exercise training decreases the expression of inflammation-related adipokines by reducing oxidative stress in WAT.

Beta2-adrenergic receptor regulate Toll-like receptor 4-induced late-phase NF-kappaB activation.

Stimulation of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) triggers myeloid differentiation factor 88 (MyD88)-dependent early-phase NF-kappaB activation and Toll/IL-1 receptor domain-containing adaptor-inducing IFN-beta (TRIF)-dependent late-phase NF-kappaB activation. In a previous study, we have shown that beta(2)-adrenergic receptor (beta(2)AR) functions as a negative regulator of NF-kappaB activation through beta-arrestin 2 in the macrophage cell line RAW264 and that down-regulation of beta(2)AR expression in response to LPS is essential for NF-kappaB activation and expression of its target gene, inducible nitric oxide synthase (NOS II). Here, we demonstrate that beta(2)AR plays an important role in TRIF-dependent late-phase NF-kappaB activation. LPS-stimulated down-regulation was induced in MyD88-knockdown cells, but not in TRIF-knockdown cells, suggesting that beta(2)AR expression was down-regulated by the TRIF-dependent pathway. On the other hand, depletion of beta(2)AR or beta-arrestin 2 expression by siRNA decreased cytoplasmic IkappaB alpha and abrogated late-phase IkappaB alpha degradation and NF-kappaB activation in response to LPS. Inducible nitric oxide synthase (NOS II) expression was increased continuously during 24 h of LPS stimulation in control cells, but decreased in beta(2)AR or beta-arrestin 2-knockdown cells after 6 h of LPS stimulation. These findings suggest that beta(2)AR functions not only as a negative regulator of NF-kappaB activation, but also as a stabilizing factor of the NF-kappaB/IkappaB alpha complex through cytoplasmic beta-arrestin 2, and that TRIF-dependent down-regulation of beta(2)AR expression increases the level of cytoplasmic NF-kappaB/IkappaB alpha complex free from beta-arrestin 2, leading to continuous late-phase NF-kappaB activation.