RESUMEN
BACKGROUND: Glucagon-like peptide-2 (GLP-2) is a pleiotropic hormone synthesized and secreted by the enteroendocrine 'L' cells able to exert intestine-trophic and anti-inflammatory effects. The antineoplastic drug cisplatin causes gastrointestinal alterations with clinical symptoms (nausea and vomiting) that greatly affect the therapy compliance. Experimentally, it has been reported that chronic cisplatin treatment caused mucosal damage and enteric neuropathy in the rat colon. METHODS: We investigated, through a combined immunohistochemical and functional approach, whether [Gly(2) ]GLP-2, a GLP-2 analog, was able to counteract the detrimental effects of long-term cisplatin administration in the mucosa and myenteric neurons of mouse gastric fundus. KEY RESULTS: Morphological experiments showed a reduction in the epithelium thickness in cisplatin-treated mice, which was prevented by [Gly(2) ]GLP-2 co-treatment. Immunohistochemistry demonstrated that cisplatin caused a significant decrease in myenteric neurons, mainly those expressing neuronal nitric oxide synthase (nNOS), that was prevented by [Gly(2) ]GLP-2 co-treatment. In the functional experiments, [Gly(2) ]GLP-2 co-treatment counteracted the increase in amplitude of the neurally induced contractions observed in strips from cisplatin-treated animals. The NO synthesis inhibitor L-N(G) -nitro arginine caused an increase in amplitude of the contractile responses that was greater in preparations from cisplatin+[Gly(2) ]GLP-2 treated mice compared to the cisplatin-treated ones. CONCLUSIONS & INFERENCES: The results demonstrate that in cisplatin long-term treated mice [Gly(2) ]GLP-2 is able to counteract both the mucosal gastric fundus damage, by preventing the epithelium thickness decrease, and the neuropathy, by protecting the nNOS neurons. Taken together, the present data suggest that [Gly(2) ]GLP-2 could represent an effective strategy to overcome the distressing gastrointestinal symptoms present during the anti-neoplastic therapy.
Asunto(s)
Antineoplásicos/toxicidad , Cisplatino/toxicidad , Péptido 2 Similar al Glucagón/farmacología , Seudoobstrucción Intestinal/inducido químicamente , Péptidos/farmacología , Animales , Western Blotting , Modelos Animales de Enfermedad , Femenino , Fundus Gástrico/efectos de los fármacos , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Plexo Mientérico/efectos de los fármacosRESUMEN
The hormone relaxin exerts a variety of functions on the smooth muscle of reproductive and nonreproductive organs, most of which occur through a nitric oxide (NO)-mediated mechanism. In the stomach and ileum, relaxin causes muscle relaxation by modulating the activity and expression of different nitric oxide synthase (NOS) isoforms region-dependently. Nothing is known on the effects of relaxin in the colon, the gut region expressing the highest number of neuronal (n) NOSß-immunoreactive neurons and mainly involved in motor symptoms of pregnancy and menstrual cycle. Therefore, we studied the effects of relaxin exposure in the mouse proximal colon in vitro evaluating muscle mechanical activity and NOS isoform expression. The functional experiments showed that relaxin decreases muscle tone and increases amplitude of spontaneous contractions; the immunohistochemical results showed that relaxin increases nNOSß and endothelial (e) NOS expression in the neurons and decreases nNOSα and eNOS expression in the smooth muscle cells (SMC). We hypothesized that, in the colon, relaxin primarily increases the activity and expression of nNOSß and eNOS in the neurons, causing a reduction of the muscle tone. The downregulation of nNOSα and eNOS expression in the SMC associated with increased muscle contractility could be the consequence of continuous exposue of these cells to the NO of neuronal origin. These findings may help to better understand the physiology of NO in the gastrointestinal tract and the role that the "relaxin-NO" system plays in motor disorders such as functional bowel disease.
Asunto(s)
Colon/metabolismo , Contracción Muscular , Músculo Liso/metabolismo , Neuronas/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Óxido Nítrico Sintasa de Tipo I/metabolismo , Relaxina/metabolismo , Anestésicos Locales/farmacología , Animales , Colon/irrigación sanguínea , Colon/citología , Colon/inervación , Colon Ascendente/citología , Colon Ascendente/efectos de los fármacos , Colon Ascendente/inervación , Colon Ascendente/metabolismo , Colon Transverso/citología , Colon Transverso/efectos de los fármacos , Colon Transverso/inervación , Colon Transverso/metabolismo , Inhibidores Enzimáticos/farmacología , Femenino , Guanilato Ciclasa/antagonistas & inhibidores , Técnicas In Vitro , Células Intersticiales de Cajal/citología , Células Intersticiales de Cajal/efectos de los fármacos , Células Intersticiales de Cajal/metabolismo , Fenómenos Mecánicos , Ratones , Ratones Endogámicos , Contracción Muscular/efectos de los fármacos , Músculo Liso/irrigación sanguínea , Músculo Liso/citología , Músculo Liso/inervación , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Proteínas del Tejido Nervioso/metabolismo , Neuronas/citología , Neuronas/efectos de los fármacos , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico Sintasa de Tipo I/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo III/antagonistas & inhibidores , Concentración Osmolar , Plexo Submucoso/citología , Plexo Submucoso/efectos de los fármacos , Plexo Submucoso/metabolismoRESUMEN
AIM: To investigate the effects of endocannabinoids on non-adrenergic, non-cholinergic (NANC) relaxant responses in gastric strips from mice. METHODS: Gastric longitudinal strips from the fundus region were mounted in organ baths for isometric recording. RESULTS: In carbachol-precontracted strips, electrical field stimulation (EFS) elicited tetrodotoxin (TTX)-sensitive fast nitrergic relaxant responses that were followed, at the highest stimulation frequency, by sustained relaxations. The latter were abolished by α-chymotrypsin. Anandamide caused a TTX-sensitive relaxation that was abolished by α-chymotrypsin but unaffected by the nitric oxide (NO) synthesis inhibitor, Nω-nitro-L-arginine (L-NNA). Anandamide reduced the amplitude of EFS-induced fast relaxations, whereas increased that of sustained ones. Relaxation to the nicotinic receptor agonist dimethylphenyl piperazinium iodide (DMPP) was decreased in amplitude by either anandamide or L-NNA, whereas, surprisingly, it was increased by α-chymotrypsin and abolished by L-NNA plus α-chymotrypsin. Relaxation to vasoactive intestinal polypeptide (VIP) was not influenced by anandamide or L-NNA and was abolished by α-chymotrypsin. Following VIP desensitization, fast relaxant responses to EFS were reduced and the sustained ones abolished. The CB1 receptor antagonist AM251 increased, only at the highest stimulation frequency, the amplitude of the EFS-induced fast relaxation and reduced the sustained one. AM251 increased the response to DMPP and abolished that to anandamide. The CB2 receptor antagonist AM630 had no effects. CONCLUSION: These results indicate that endocannabinoids modulate, via prejunctional CB1 receptors, the NANC peptidergic neurotransmission that, in turn, affects the nitrergic one.
Asunto(s)
Endocannabinoides/metabolismo , Contracción Muscular/efectos de los fármacos , Estómago/efectos de los fármacos , Estómago/inervación , Transmisión Sináptica/fisiología , Animales , Agonistas de Receptores de Cannabinoides/farmacología , Antagonistas de Receptores de Cannabinoides/farmacología , Femenino , Ratones , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/fisiología , Receptores de Cannabinoides/metabolismoRESUMEN
Impaired gastric motility ascribable to a defective nitric oxide (NO) production has been reported in dystrophic (mdx) mice. Since relaxin upregulates NO biosynthesis, its effects on the motor responses and NO synthase (NOS) expression in the gastric fundus of mdx mice were investigated. Mechanical responses of gastric strips were recorded via force displacement transducers. Evaluation of the three NOS isoforms was performed by immunohistochemistry and Western blot. Wild-type (WT) and mdx mice were distributed into three groups: untreated, relaxin pretreated, and vehicle pretreated. In strips from both untreated and vehicle-pretreated animals, electrical field stimulation (EFS) elicited contractile responses that were greater in mdx than in WT mice. In carbachol-precontracted strips, EFS induced fast relaxant responses that had a lower amplitude in mdx than in WT mice. Only in the mdx mice did relaxin depress the amplitude of the neurally induced excitatory responses and increase that of the inhibitory ones. In the presence of L-NNA, relaxin was ineffective. In relaxin-pretreated mdx mice, the amplitude of the EFS-induced contractile responses was decreased and that of the fast relaxant ones was increased compared with untreated mdx animals. Responses to methacholine or papaverine did not differ among preparations and were not influenced by relaxin. Immunohistochemistry and Western blotting showed a significant decrease in neuronal NOS expression and content in mdx compared with WT mice, which was recovered in the relaxin-pretreated mdx mice. The results suggest that relaxin is able to counteract the altered contractile and relaxant responses in the gastric fundus of mdx mice by upregulating nNOS expression.