Effects of ghrelin on neuronal survival in cells derived from dorsal motor nucleus of the vagus.
BACKGROUND
The effects of intestinal inflammation on the central neurons projecting to the enteric nervous system are unknown. The dorsal motor nucleus of the vagus signals to the gastrointestinal system. Ghrelin is elevated in patients with inflammatory bowel disease and has been implicated as an inflammatory mediator. The purpose of this study was to investigate the effects of gastrointestinal inflammation on the dorsal motor nucleus of the vagus in rats, as well as the effects of proinflammatory cytokines and ghrelin on neurons from the dorsal motor nucleus of the vagus in vitro.


METHODS
DiI was injected into the stomach wall of rats to retrogradely label neurons of the dorsal motor nucleus of the vagus. Intestinal inflammation was induced with indomethacin injection. Serial serum ghrelin measurements were performed. Tissue was examined under fluorescent microscopy. In vitro studies using primary culture of neurons from the dorsal motor nucleus of the vagus were performed. Reverse transcriptase-polymerase chain reaction for cytokine transcripts and immunohistochemistry for cytokine receptors were performed. Cell proliferation and apoptosis were measured by enzyme-linked immunosorbent assay.


RESULTS
A significant decrease of DiI labeling was demonstrated in the dorsal motor nucleus of the vagus of animals injected with indomethacin. Serum levels of ghrelin were significantly elevated 2 days after induction of inflammation. In vitro, apoptosis and cell proliferation were measured after 24-hour exposure to experimental conditions. Ghrelin alone had no effect on apoptosis. Exposure to interleukin (IL)-1 beta or tumor necrosis factor (TNF)-alpha increased apoptosis. The addition of ghrelin to cytokine resulted in significant decreases in apoptosis compared to cytokine alone. Ghrelin significantly increased neuronal proliferation. Exposure to IL-1 beta, IL-6, or TNF-alpha significantly decreased proliferation. The addition of ghrelin to TNF-alpha or IL-6 significantly increased cellular proliferation compared to cytokine alone.


CONCLUSIONS
Neurons from the dorsal motor nucleus of the vagus that project to the stomach are reduced in number after induction of colitis in rats. In vitro, proinflammatory cytokines increase apoptosis and decrease cell proliferation of neurons from the dorsal motor nucleus of the vagus. These effects are attenuated by ghrelin.
Ammori
John B
JB
Department of Surgery, University of Michigan, Ann Arbor, MI 48109-0346, USA.
Zhang
Wei-Zhen
WZ
Li
Ji-Yao
JY
Chai
Biao-Xin
BX
Mulholland
Michael W
MW
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003092:Colitis / Q000139:chemically induced / Q000473:pathology*; D041981:Gastrointestinal Tract / Q000294:innervation; D054439:Ghrelin / Q000097:blood / Q000494:pharmacology*; D007213:Indomethacin; D008297:Male; D008526:Medulla Oblongata / Q000187:drug effects / Q000473:pathology; D009434:Neural Pathways; D009476:Neurons, Efferent / Q000737:chemistry / Q000187:drug effects* / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017472:Receptors, Interleukin-1 / Q000032:analysis; D019947:Receptors, Interleukin-6 / Q000032:analysis; D014302:Trinitrobenzenesulfonic Acid; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis; D014630:Vagus Nerve / Q000187:drug effects* / Q000473:pathology
D054439:Ghrelin; D017472:Receptors, Interleukin-1; D019947:Receptors, Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D014302:Trinitrobenzenesulfonic Acid; D007213:Indomethacin