Yuichi Oishi

Effect of exercise intensity and AICAR on isoform-specific expressions of murine skeletal muscle PGC-1α mRNA: a role of β2-adrenergic receptor activation

There are three isoforms of peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) mRNA, which promotes mitochondrial biogenesis in skeletal muscles. Compared with PGC-1α-a mRNA, PGC-1α-b or PGC-1α-c mRNA is transcribed by a different exon 1 of the PGC-1α gene. In this study, effects of exercise intensity and 5-aminoimidazole-4-carboxamide-1β-d-ribofuranoside (AICAR) on isoform-specific expressions of PGC-1α were investigated. All isoforms were increased in proportion to exercise intensity of treadmill running (10-30 m/min for 30 min). Preinjection of β₂-adrenergic receptor (AR) antagonist (ICI 118551) inhibited the increase in PGC-1α-b and PGC-1α-c mRNAs, but not the increase in PGC-1α-a mRNA, in response to high-intensity exercise. Although high-intensity exercise activated α2-AMP-activated protein kinase (α2-AMPK) in skeletal muscles, inactivation of α2-AMPK activity did not affect high-intensity exercise-induced mRNA expression of all PGC-1α isoforms, suggesting that activation of α2-AMPK is not mandatory for an increase in PGC-1α mRNA by high-intensity exercise. A single injection in mice of AICAR, an AMPK activator, increased mRNAs of all PGC-1α isoforms. AICAR increased blood catecholamine concentrations, and preinjection of β₂-AR antagonist inhibited the increase in PGC-1α-b and PGC-1α-c mRNAs but not the increase in PGC-1α-a mRNA. Direct exposure of epitrochlearis muscle to AICAR increased PGC-1α-a but not the -b isoform. These data indicate that exercise-induced PGC-1α expression was dependent on the intensity of exercise. Exercise or AICAR injection increased PGC-1α-b and PGC-1α-c mRNAs via β₂-AR activation, whereas high-intensity exercise increased PGC-1α-a expression by a multiple mechanism in which α2-AMPK is one of the signaling pathways.

Skeletal Muscle-Specific Expression of PGC-1α-b, an Exercise-Responsive Isoform, Increases Exercise Capacity and Peak Oxygen Uptake

Background Maximal oxygen uptake (VO2max) predicts mortality and is associated with endurance performance. Trained subjects have a high VO2max due to a high cardiac output and high metabolic capacity of skeletal muscles. Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), a nuclear receptor coactivator, promotes mitochondrial biogenesis, a fiber-type switch to oxidative fibers, and angiogenesis in skeletal muscle. Because exercise training increases PGC-1α in skeletal muscle, PGC-1α-mediated changes may contribute to the improvement of exercise capacity and VO2max. There are three isoforms of PGC-1α mRNA. PGC-1α-b protein, whose amino terminus is different from PGC-1α-a protein, is a predominant PGC-1α isoform in response to exercise. We investigated whether alterations of skeletal muscle metabolism by overexpression of PGC-1α-b in skeletal muscle, but not heart, would increase VO2max and exercise capacity. Methodology/Principal Findings Transgenic mice showed overexpression of PGC-1α-b protein in skeletal muscle but not in heart. Overexpression of PGC-1α-b promoted mitochondrial biogenesis 4-fold, increased the expression of fatty acid transporters, enhanced angiogenesis in skeletal muscle 1.4 to 2.7-fold, and promoted exercise capacity (expressed by maximum speed) by 35% and peak oxygen uptake by 20%. Across a broad range of either the absolute exercise intensity, or the same relative exercise intensities, lipid oxidation was always higher in the transgenic mice than wild-type littermates, suggesting that lipid is the predominant fuel source for exercise in the transgenic mice. However, muscle glycogen usage during exercise was absent in the transgenic mice. Conclusions/Significance Increased mitochondrial biogenesis, capillaries, and fatty acid transporters in skeletal muscles may contribute to improved exercise capacity via an increase in fatty acid utilization. Increases in PGC-1α-b protein or function might be a useful strategy for sedentary subjects to perform exercise efficiently, which would lead to prevention of life-style related diseases and increased lifespan.

Inhibition of increases in blood glucose and serum neutral fat by Momordica charantia saponin fraction.

Focusing on a functional component of Momordica charantia, saponin, we investigated its effects on serum glucose and neutral fat levels. Saponin was extracted as a butanol-soluble fraction (saponin fraction) from hot blast-dried Momordica charantia powder. The disaccharidase-inhibitory activity and the pancreatic lipase-inhibitory activity of the saponin fraction were measured, and in vivo sugar- and lipid-loading tests were performed. The saponin fraction inhibited disaccharidase activity and elevation of the blood glucose level after sucrose loading. The fraction also markedly inhibited pancreatic lipase activity and elevation of the serum neutral fat level after corn oil loading. Based on these findings, the main active component related to the anti-diabetic effect of Momordica charantia is present in the butanol fraction, and it may be saponin. The blood glucose and serum neutral fat-lowering effects of Momordica charantia were closely associated with its inhibitory activity against disaccharidase and pancreatic lipase.

The hypocholesterolemic activity of Momordica charantia fruit is mediated by the altered cholesterol- and bile acid–regulating gene expression in rat liver

Although many studies have demonstrated the hypocholesterolemic activity of Momordica charantia, also called bitter gourd fruit (BGF), the relative hypocholesterolemic mechanism is not fully understood. In the present study, we hypothesized that BGF alters hepatic gene expression of cholesterol- and bile acid-regulating proteins to improve blood cholesterol profiles. To clarify the mechanism, we fed 7-week-old male Wistar rats a high-cholesterol (HC) diet containing 5% BGF for 4 weeks and determined the cholesterol levels in the serum, liver and feces, concentrations of the fecal total bile acid, and the expression level of cholesterol- and bile acid-regulating genes. The HC diet with BGF supplementation showed a significant serum hypocholesterolemic activity compared with the HC diet without BGF. BGF intake also significantly increased the levels of fecal total bile acid, suggesting that BGF inhibited the reabsorption of bile acids into the intestine. Hepatic messenger RNA (mRNA) levels of small heterodimer partner (SHP) and liver receptor homolog-1, which are both involved in cholesterol 7α-hydroxylase (CYP7A1) regulation, were significantly decreased and increased, respectively, by BGF intake. In addition, BGF tended to increase the hepatic CYP7A1 mRNA level. Taken together, these results suggest that BGF not only decreases the reabsorption of bile acids into the intestine but also increases the conversion of cholesterol to bile acids by CYP7A1 up-regulation through the down-regulation of the hepatic farnesoid X receptor/SHP pathway.

Adiponectin promotes hyaluronan synthesis along with increases in hyaluronan synthase 2 transcripts through an AMP-activated protein kinase/peroxisome proliferator-activated receptor-α-dependent pathway in human dermal fibroblasts

Although adipocytokines affect the functions of skin, little information is available on the effect of adiponectin on the skin. In this study, we investigated the effect of adiponectin on hyaluronan synthesis and its regulatory mechanisms in human dermal fibroblasts. Adiponectin promoted hyaluronan synthesis along with an increase in the mRNA levels of hyaluronan synthase 2 (HAS2), which plays a primary role in hyaluronan synthesis. Adiponectin also increased the phosphorylation of AMP-activated protein kinase (AMPK). A pharmacological activator of AMPK, 5-aminoimidazole-4-carboxamide-1β-ribofuranoside (AICAR), increased mRNA levels of peroxisome proliferator-activated receptor-α (PPARα), which enhances the expression of HAS2 mRNA. In addition, AICAR increased the mRNA levels of HAS2. Adiponectin-induced HAS2 mRNA expression was blocked by GW6471, a PPARα antagonist, in a concentration-dependent manner. These results show that adiponectin promotes hyaluronan synthesis along with increases in HAS2 transcripts through an AMPK/PPARα-dependent pathway in human dermal fibroblasts. Thus, our study suggests that adiponectin may be beneficial for retaining moisture in the skin, anti-inflammatory activity, and the treatment of a variety of cutaneous diseases.

A high-fat diet reduces ceramide synthesis by decreasing adiponectin levels and decreases lipid content by modulating HMG-CoA reductase and CPT-1 mRNA expression in the skin.

SCOPE Molecules involved in skin function are greatly affected by nutritional conditions. However, the mechanism linking high-fat (HF) diets with these alterations is not well understood. This study aimed to investigate the molecular changes in skin function that result from HF diets. METHODS AND RESULTS Sprague-Dawley rats were fed HF diets for 28 days. The skin levels of ceramide, lipids and mRNAs involved in lipid metabolism were evaluated using TLC, oil red O staining and quantitative PCR, respectively. The serum adiponectin concentration was determined by ELISA. HF diets led to reduced ceramide levels and lowered skin lipid content. They also decreased mRNA levels of serine palmitoyltransferase (SPT) and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in the skin and those of peroxisome proliferator-activated receptor-α -PPAR-α), which upregulates SPT and HMG-CoA reductase expression. The HF diets reduced the serum concentration of adiponectin, which acts upstream of PPAR-α. Finally, these diets led to increased mRNA levels of carnitine palmitoyltransferase-1, the rate-limiting enzyme that acts in β-oxidation. CONCLUSION Our study suggests that HF diets reduce ceramide and lipid synthesis in the skin by reducing levels of SPT and HMG-CoA reductase through lowered adiponectin and PPAR-α activity. Additionally, they decrease lipid content by enhancing β-oxidation.

High-fat diet reduces levels of type I tropocollagen and hyaluronan in rat skin.

Although it is known that nutritional conditions affect the skin function, little information is available on the effect of a high-fat (HF) diet on skin. In this study, Sprague-Dawley rats were fed HF diets for 28 days, and we investigated the effect of this diet on type I tropocollagen and hyaluronan in rat skin. The HF diets reduced the levels of type I tropocollagen, COL1A1 mRNA, hyaluronan, and rat hyaluronan synthase (rhas)2 mRNA, which play a primary role in hyaluronan synthase in the dermis. However, rhas3 mRNA level in the skin was increased. The HF diets also decreased the skin mRNA expression of transforming growth factor (TGF)-beta1, which enhances the expression of COL1A1 and rhas2 mRNA and decreases rhas3 mRNA expression, and decreased the hepatic mRNA expression of insulin-like growth factor (IGF)-I, which enhances COL1A1, rhas2, and TGF-beta1 mRNA expression. The serum level of adiponectin, which promotes the syntheses of type I collagen and hyaluronan, was decreased in the HF diet groups. These findings suggest that an HF diet reduces the levels of type I tropocollagen and hyaluronan in the skin by suppressing the action of TGF-beta1, IGF-I and adiponectin, and these effects are deleterious for skin function.

Hydrocellular foam dressing promotes wound healing along with increases in hyaluronan synthase 3 and PPARα gene expression in epidermis.

Background Hydrocellular foam dressing, modern wound dressing, induces moist wound environment and promotes wound healing: however, the regulatory mechanisms responsible for these effects are poorly understood. This study was aimed to reveal the effect of hydrocellular foam dressing on hyaluronan, which has been shown to have positive effects on wound healing, and examined its regulatory mechanisms in rat skin. Methodology/Principal Findings We created two full-thickness wounds on the dorsolateral skin of rats. Each wound was covered with either a hydrocellular foam dressing or a film dressing and hyaluronan levels in the periwound skin was measured. We also investigated the mechanism by which the hydrocellular foam dressing regulates hyaluronan production by measuring the gene expression of hyaluronan synthase 3 (Has3), peroxisome proliferator-activated receptor α (PPARα), and CD44. Hydrocellular foam dressing promoted wound healing and upregulated hyaluronan synthesis, along with an increase in the mRNA levels of Has3, which plays a primary role in hyaluronan synthesis in epidermis. In addition, hydrocellular foam dressing enhanced the mRNA levels of PPARα, which upregulates Has3 gene expression, and the major hyaluronan receptor CD44. Conclusions/Significance These findings suggests that hydrocellular foam dressing may be beneficial for wound healing along with increases in hyaluronan synthase 3 and PPARα gene expression in epidermis. We believe that the present study would contribute to the elucidation of the mechanisms underlying the effects of hydrocellular foam dressing-induced moist environment on wound healing and practice evidence-based wound care.

Serum Cholesterol-Decreasing Effect of Heat-Moisture-Treated High-Amylose Cornstarch in Cholesterol-Loaded Rats

Rats were fed on a diet containing cholesterol (Chol) at a level corresponding to the standard Chol intake in humans, and the influence of heat-moisture-treated high-amylose cornstarch (HHA) on their serum Chol level was investigated. HHA decreased the serum level of Chol in rats fed on the diet containing 0.1% Chol, which corresponds to a Chol intake in humans of 800 mg/d, although the liver levels of Chol increased in these rats. HHA did not influence the fecal excretion of Chol/bile acids. It is possible that the decrease in serum Chol level in the rats fed on the high-Chol diet can be attributed to the promotion of Chol uptake in the liver.

Hydrocellular foam dressings promote wound healing associated with decrease in inflammation in rat periwound skin and granulation tissue, compared with hydrocolloid dressings

The effects of modern dressings on inflammation, which represent the earliest phase of wound healing, are poorly understood. We investigated the effects of modern hydrocellular foam dressings (HCFs) on wound healing and on the gene expression levels of the inflammatory markers—interleukin (IL)-1β, IL-6, and IL-10—in rat periwound skin and granulation tissue by quantitative reverse transcription-polymerase chain reaction. HCF absorbed significantly higher volume of water than hydrocolloid dressing (HCD) and increased the contraction of wounds. Polymorphonuclear neutrophils were massively infiltrated to the wound edge and boarded between granulation and dermis in the HCD group. IL-1β, IL-6, and IL-10 mRNA levels were significantly decreased in the periwound skin around the wounds and granulation tissue covered with HCF. These findings suggest that HCF may promote wound healing along with decrease in inflammation by reducing gene expression levels of IL-1β, IL-6, and IL-10. Graphic·Abstract HCF promotes wound healing along with decreases in inflammation by reducing gene expression levels of IL-1β, IL-6 and IL-10 in rat periwound skin and granulation tissue.