Hidesuke Kaji

Chemerin enhances insulin signaling and potentiates insulin‐stimulated glucose uptake in 3T3‐L1 adipocytes

To explore a novel adipokine, we screened adipocyte differentiation‐related gene and found that TIG2/chemerin was strongly induced during the adipocyte differentiation. Chemerin was secreted by the mature 3T3‐L1 adipocytes and expressed abundantly in adipose tissue in vivo as recently described. Intriguingly, the expression of chemerin was differently regulated in the liver and adipose tissue in db/db mice. In addition, serum chemerin concentration was decreased in db/db mice. Chemerin and its receptor/ChemR23 were expressed in mature adipocytes, suggesting its function in autocrine/paracrine fashion. Finally, chemerin potentiated insulin‐stimulated glucose uptake concomitant with enhanced insulin signaling in the 3T3‐L1 adipocytes. These data establish that chemerin is a novel adipokine that regulates adipocyte function.

Neither intravenous nor intracerebroventricular administration of obestatin affects the secretion of GH, PRL, TSH and ACTH in rats

To examine the effect of obestatin, a recently identified peptide derived from preproghrelin, on pituitary hormone secretion, obestatin was administered in anesthetized male rats. Intravenous administration of obestatin did not show any effect on plasma GH, PRL, ACTH and TSH levels. Since obestatin has been reported to have opposite effects of ghrelin in regulating food intake, gastric emptying and intestinal contractility, GH suppressive effect, which is opposite effect of ghrelin, was tested. Intravenous administration of GHRH or GHRP-2, a ghrelin receptor ligand, resulted in a marked plasma GH elevation. However obestatin did not show any effect on GHRH- or GHRP-2-induced GH rise. Furthermore intracerebroventricular administration of obestatin also did not influence plasma GH, PRL, ACTH and TSH levels. These findings suggest that obestatin has no effect on pituitary hormone secretions despite the presence of GPR39, a receptor for obestatin, in the pituitary.

Leptin Stimulates Basal and GHRH-Induced GH Release from Cultured Rat Anterior Pituitary Cells in Vitro

Leptin, the product of ob gene, is secreted from adipocytes and appears to regulate food intake and energy expenditure through its receptor in the hypothalamus. In addition, leptin is reported to modulate pituitary hormones including LH, FSH and ACTH, probably via the hypothalamus. In obesity, growth hormone (GH) secretion is impaired, while serum leptin levels are elevated. To investigate the possibility that leptin serves as a metabolic signal that influences GH secretion from the pituitary, we studied the effect of leptin on GH secretion from primary monolayer cultures of rat anterior pituitary cells. Twenty micrograms/ml leptin increased GH secretion, but did not increase GH release at a physiological concentration up to 200 ng/ml. However, 200 ng/ml leptin stimulated GH release in the presence of GH-releasing hormone (GHRH) ranging from 10(-8) to 10(-7) M. Twenty ng/ml leptin tended to increase 10(-7) M GHRH-induced GH release, whereas 20 ng/ml leptin did not increase either 10(-9) or 10(-8) M GHRH-induced GH release. This result suggests that leptin has a direct effect on the pituitary to enhance GHRH-induced GH secretion.

GHRP-2, a GHS-R agonist, directly acts on myocytes to attenuate the dexamethasone-induced expressions of muscle-specific ubiquitin ligases, Atrogin-1 and MuRF1.

Recent reports suggest that Atrogin-1 and MuRF1, E3 ubiquitin ligases, play a pivotal role in muscle atrophy. In the present study, effect of Growth Hormone Releasing Peptide-2 (GHRP-2), a GH secretagogue receptor (GHS-R) agonist, on the expressions of Atrogin-1 and MuRF1 in vivo rat muscles was examined. Dexamethasone administration increased Atrogin-1 mRNA level in rat soleus muscle. The increased mRNA level of Atrogin-1 was significantly attenuated by GHRP-2. In addition, GHRP-2 decreased MuRF1 mRNA level irrespective of the presence of dexamethasone. Although IGF-I is a well-known protective factor for muscle atrophy, GHRP-2 did not influence plasma IGF-I levels and IGF-I mRNA levels in muscles. To clarify a direct effect of GHRP-2, differentiated C2C12 myocytes were used. Ten micrometer dexamethasone increased both Atrogin-1 and MuRF1 mRNA levels in C2C12 cells. GHRP-2 attenuated dexamethasone-induced expression of them dose-dependently and decreased the basal level of MuRF1 mRNA. The suppressive effect on the expressions of Atrogin-1 and MuRF1 by GHRP-2 was blocked by [D-Lys(3)]-GHRP-6, a GHS-R1a blocker, suggesting the effect of GHRP-2 was mediated through GHS-R1a. Taken together, GHRP-2 directly attenuates Atrogin-1 and MuRF1 mRNA levels through ghrelin receptors in myocytes.

Low Serum IGF-I/GH Ratio Is Associated with Abnormal Glucose Tolerance in Acromegaly

Background/Aims: Acromegaly is frequently accompanied with impaired glucose tolerance (IGT) and diabetes mellitus (DM). It remains unclear which factors determine the abnormal glucose tolerance status in acromegaly. In addition, diverse actions of GH and IGF-I in regulating glucose metabolism in acromegaly have not yet been well elucidated. The aim of this study was to investigate the factors associated with abnormal glucose tolerance in acromegaly. Subjects and Study Design: We conducted a retrospective cross-sectional study that included 48 patients with active acromegaly. The subjects were divided into two groups by the results of 75 g OGTT: normal glucose tolerance (NGT) group (n = 19) and IGT+DM group (n = 29). Results: Systolic blood pressure (SBP) was significantly higher in the IGT+DM than in the NGT group. Homeostasis model assessment of β-cell function (HOMA-β) was significantly decreased in the IGT+DM group compared with the NGT group. Although serum GH or IGF-I levels were not different between the two groups, the IGF-I/GH ratio in the IGT+DM group was significantly lower than that in the NGT group. Conclusions: We have shown that a low serum IGF-I/GH ratio was associated with abnormal glucose tolerance in acromegaly. We propose that the IGF-I/GH ratio is a useful marker to understand the metabolic status in acromegaly.

Involvement of mPOU (Brn-5), a class VI POU protein, in the gene expression of Pit-1 as well as PRL.

PRL is mainly expressed in the pituitary and its gene expression is regulated by a variety of transcription factors including Pit-1. Brn-5 is a transcription factor that binds to Pit-1 binding elements and stimulates PRL reporter gene expression. In this study, the role of Brn-5 was examined. RNA interference (RNAi) against Brn-5 leaded to reduction in PRL content of GH3 cells, indicating endogenous Brn-5 may play a role in PRL gene expression. Furthermore Brn-5 RNAi decreased Pit-1 mRNA. Transfection of expression vectors for mPOU (human ortholog of Brn-5) modestly but significantly stimulated activities of PRL-Luc and Pit-1-Luc reporter genes in GH3 and HEK 293 cells. In addition, mPOU showed synergistic action with Pit-1 and CBP on PRL-Luc expression. mPOU-FL, a splicing variant of mPOU, showed weaker activity than mPOU. Chip assay suggested binding of mPOU to PRL and Pit-1 promoters of genomic DNA. Taken together, these results suggest that mPOU (Brn-5) enhances PRL gene expression directly in association with Pit-1 and CBP, and indirectly via the activation of Pit-1 gene expression.

Genetic Variations at the CCAAT/Enhancer-binding Protein δ are Associated with Metabolic Phenotypes in the Japanese Population

BACKGROUNDnExpression of CCAAT/enhancer-binding protein delta (C/EBP-delta) gene is enhanced in the early initial stage of adipocyte differentiation. This study is intended to elucidate the association between the genetic variation of C/EBP-delta and metabolic phenotypes.nnnSUBJECTS AND METHODSnSubjects were unselected 52 males and 120 females in Japan, aged 40 to 79 years, who visited a city hygienic center for a health checkup and agreed to participation in the study after giving informed consent. The C/EBP-delta genotypes and metabolic phenotypes were determined. An association study was performed using Pearsons chi(2) tests and logistic regression analyses.nnnRESULTSnTwo SNPs in C/EBP-delta gene with minor allele frequency greater than 0.05 were detectable among seven SNPs. Genotype heterozygous for the C and T allele (877C/T) were more prevalent in subjects with dyslipidemia [plasma triglyceride (TG) > or =150 mg/dL and/or plasma high-density lipoprotein-cholesterol (HDL-C) <40 mg/dL] as well as high fasting plasma glucose (FPG; > or =110 mg/dL) than controls (22.5% vs. 7.6%, P = 0.009, and 20.0% vs. 8.4%, P = 0.041, respectively). Moreover, the genotype 877C/T contributed to both dyslipidemia and high FPG independent of age, sex, and visceral obesity. Regarding 394C>G, no association between the genotype and metabolic phenotypes was detected.nnnCONCLUSIONnResults suggest that genetic variations in the C/EBP-delta might play a role in some metabolic phenotypes.