Acylated and unacylated ghrelin promote proliferation and inhibit apoptosis of pancreatic beta-cells and human islets: involvement of 3',5'-cyclic adenosine monophosphate/protein kinase A, extracellular signal-regulated kinase 1/2, and phosphatidyl inositol 3-Kinase/Akt signaling.

• Among its pleiotropic actions, ghrelin modulates insulin secretion and glucose metabolism. Herein we investigated the role of ghrelin in pancreatic beta-cell proliferation and apoptosis induced by serum starvation or interferon (IFN)-gamma/TNF-alpha, whose synergism is a major cause for beta-cell destruction in type I diabetes. HIT-T15 beta-cells expressed ghrelin but not ghrelin receptor (GRLN-R), which binds acylated ghrelin (AG) only. However, both unacylated ghrelin (UAG) and AG recognized common high-affinity binding sites on these cells. Either AG or UAG stimulated cell proliferation through Galpha(s) protein and prevented serum starvation- and IFN-gamma/TNF-alpha-induced apoptosis. Antighrelin antibody enhanced apoptosis in either the presence or absence of serum but not cytokines. AG and UAG even up-regulated intracellular cAMP. Blockade of adenylyl cyclase/cAMP/protein kinase A signaling prevented the ghrelin cytoprotective effect. AG and UAG also activated phosphatidyl inositol 3-kinase (PI3K)/Akt and ERK1/2, whereas PI3K and MAPK inhibitors counteracted the ghrelin antiapoptotic effect. Furthermore, AG and UAG stimulated insulin secretion from HIT-T15 cells. In INS-1E beta-cells, which express GRLN-R, AG and UAG caused proliferation and protection against apoptosis through identical signaling pathways. Noteworthy, both peptides inhibited cytokine-induced NO increase in either HIT-T15 or INS-1E cells. Finally, they induced cell survival and protection against apoptosis in human islets of Langerhans. These expressed GRLN-R but showed also UAG and AG binding sites. Our data demonstrate that AG and UAG promote survival of both beta-cells and human islets. These effects are independent of GRLN-R, are likely mediated by AG/UAG binding sites, and involve cAMP/PKA, ERK1/2, and PI3K/Akt.

Nyckelord

1-Phosphatidylinositol 3-Kinase

metabolism

Acylation

Animals

Apoptosis

drug effects

Binding Sites

Cell Line

Cell Proliferation

drug effects

Cricetinae

Culture Media

Serum-Free

pharmacology

Cyclic AMP

physiology

Cyclic AMP-Dependent Protein Kinases

physiology

Drug Synergism

Extracellular Signal-Regulated MAP Kinases

physiology

GTP-Binding Protein alpha Subunits

Gs

metabolism

Humans

Insulin

secretion

Insulin-Secreting Cells

cytology

physiology

secretion

Interferon Type II

pharmacology

Islets of Langerhans

cytology

physiology

Nitric Oxide

antagonists & inhibitors

biosynthesis

Peptide Hormones

chemistry

pharmacology

Proto-Oncogene Proteins c-akt

metabolism

Receptors

G-Protein-Coupled

metabolism

Signal Transduction

physiology

Tumor Necrosis Factor-alpha

pharmacology

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