ABSTRACT
AIM: To determine whether Nucb2/nesfatin1 production is regulated by the cannabinoid system through the intracellular mTOR pathway in the stomach. METHODS: Sprague Dawley rats were treated with vehicle, rimonabant, rapamycin or rapamycin+rimonabant. Gastric tissue obtained from the animals was used for biochemical assays: Nucb2 mRNA measurement by real time PCR, gastric Nucb2/nesfatin protein content by western blot, and gastric explants to obtain gastric secretomes. Nucb2/nesfatin levels were measured in gastric secretomes and plasma using enzyme-linked immunosorbent assay. RESULTS: The inhibition of cannabinoid receptor 1 (CB1) by the peripheral injection of an inverse agonist, namely rimonabant, decreases food intake and increases the gastric secretion and circulating levels of Nucb2/nesfatin-1. In addition, rimonabant treatment activates mTOR pathway in the stomach as showed by the increase in pmTOR/mTOR expression in gastric tissue obtained from rimonabant treated animals. These effects were confirmed by the use of a CB1 antagonist, AM281. When the intracellular pathway mTOR/S6k was inactivated by chronic treatment with rapamycin, rimonabant treatment was no longer able to stimulate the gastric secretion of Nucb2/nesfatin-1. CONCLUSION: The peripheral cannabinoid system regulates food intake through a mechanism that implies gastric production and release of Nucb2/Nesfatin-1, which is mediated by the mTOR/S6k pathway.
Subject(s)
Calcium-Binding Proteins/metabolism , Cannabinoid Receptor Antagonists/pharmacology , DNA-Binding Proteins/metabolism , Nerve Tissue Proteins/metabolism , Receptor, Cannabinoid, CB1/antagonists & inhibitors , Signal Transduction/drug effects , TOR Serine-Threonine Kinases/metabolism , Animals , Calcium-Binding Proteins/blood , DNA-Binding Proteins/blood , Eating/drug effects , Enzyme-Linked Immunosorbent Assay , Gastric Mucosa/drug effects , Gastric Mucosa/metabolism , Gastric Mucosa/pathology , Male , Models, Animal , Morpholines/pharmacology , Nerve Tissue Proteins/blood , Nucleobindins , Phosphorylation , Piperidines/pharmacology , Pyrazoles/pharmacology , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Ribosomal Protein S6 Kinases/metabolism , Rimonabant , Sirolimus/pharmacology , TOR Serine-Threonine Kinases/antagonists & inhibitorsABSTRACT
The secretion of the hepatokine alpha-2-Heremans-Schmid glycoprotein/Fetuin A, implicated in pathological processes including systemic insulin resistance, by adipose tissue has been recently described. Thus, we have recently identified its presence in white adipose tissue secretomes by mass spectrometry. However, the secretion pattern and function of adipose-derived alpha-2-Heremans-Schmid glycoprotein are poorly understood. The aim of this study is to evaluate the expression and secretion of total and active phosphorylated alpha-2-Heremans-Schmid glycoprotein by adipose tissue from visceral and subcutaneous localizations in animals at different physiological and nutritional status including anorexia and obesity. Alpha-2-Heremans-Schmid glycoprotein expression and secretion in visceral adipose tissue and subcutaneous adipose tissue explants from animals under fasting and exercise training, at pathological situations such as anorexia and obesity, and from human obese individuals were assayed by immunoblotting, quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. We reveal that visceral adipose tissue expresses and secretes more alpha-2-Heremans-Schmid glycoprotein than subcutaneous adipose tissue, and that this secretion is diminished after fasting and exercise training. Visceral adipose tissue from anorectic animals showed reduced alpha-2-Heremans-Schmid glycoprotein secretion; on the contrary, alpha-2-Heremans-Schmid glycoprotein is over-secreted by visceral adipose tissue in the occurrence of obesity. While secretion of active-PhophoSer321α2HSG by visceral adipose tissue is independent of body mass index, we found that the fraction of active-alpha-2-Heremans-Schmid glycoprotein secreted by subcutaneous adipose tissue increments significantly in situations of obesity. Functional studies show that the inhibition of adipose-derived alpha-2-Heremans-Schmid glycoprotein increases insulin sensitivity in differentiated adipocytes. In conclusion, visceral adipose tissue secretes more alpha-2-Heremans-Schmid glycoprotein than subcutaneous adipose tissue and this secretion is more sensitive to nutritional and physiological changes. The over-secretion of alpha-2-Heremans-Schmid glycoprotein by visceral adipose tissue, the increased secretion of the active phosphorylated form by subcutaneous adipose tissuein obese animals, and the adipose-derived alpha-2-Heremans-Schmid glycoprotein capacity to inhibit the insulin pathway suggest the participation of adipose-derived alpha-2-Heremans-Schmid glycoprotein in the deleterious effects of obesity.
Subject(s)
Intra-Abdominal Fat/metabolism , Obesity/metabolism , Subcutaneous Fat/metabolism , alpha-2-HS-Glycoprotein/metabolism , Adipocytes/drug effects , Adipocytes/metabolism , Animals , Cells, Cultured , Female , Humans , Insulin/metabolism , Insulin/pharmacology , Insulin Resistance/physiology , Male , Phosphorylation , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effectsABSTRACT
In the context of obesity, strong evidences support a distinctive pathological contribution of adipose tissue depending on its anatomical site of accumulation. Therefore, subcutaneous adipose tissue (SAT) has been lately considered metabolically benign compared to visceral fat (VAT), whose location is associated to the risk of developing cardiovascular disease, insulin resistance, and other associated comorbidities. Under the above situation, the chronic local inflammation that characterizes obese adipose tissue, has acquired a major role on the pathogenesis of obesity. In this work, we have analyzed for the first time human obese VAT and SAT secretomes using an improved quantitative proteomic approach for the study of tissue secretomes, Comparison of Isotope-Labeled Amino acid Incorporation Rates (CILAIR). The use of double isotope-labeling-CILAIR approach to analyze VAT and SAT secretomes allowed the identification of location-specific secreted proteins and its differential secretion. Additionally to the very high percentage of identified proteins previously implicated in obesity or in its comorbidities, this approach was revealed as a useful tool for the study of the obese adipose tissue microenvironment including extracellular matrix (ECM) remodeling and inflammatory status. The results herein presented reinforce the fact that VAT and SAT depots have distinct features and contribute differentially to metabolic disease.
Subject(s)
Adipose Tissue/metabolism , Extracellular Matrix/metabolism , Inflammation/metabolism , Intra-Abdominal Fat/metabolism , Obesity/metabolism , Subcutaneous Fat/metabolism , Amino Acids/metabolism , Humans , Inflammation Mediators/metabolism , Insulin Resistance/physiology , Isotope Labeling/methods , Proteomics/methodsABSTRACT
Nesfatin-1, which is derived from the NEFA/nucleobindin 2 (NUCB2) precursor, was recently identified as an anorexigenic peptide that is produced in several tissues including the hypothalamus. Currently, no data exist regarding the regulation of NUCB2/nesfatin-1 production in peripheral tissues, such as gastric mucosa and adipose tissue, through different periods of development. The aim of the present work was to study the variations on circulating levels, mRNA expression and tissue content in gastric mucosa and adipose tissue of NUCB2/nesfatin-1 with age and specially in two clue periods of maturation, weaning and puberty. The weaning period affected NUCB2/nesfatin-1 production in gastric tissue. The testosterone changes associated with the initiation of puberty regulated NUCB2/nesfatin-1 production via adipose tissue and gastric NUCB2/nesfatin-1 production. In conclusion, the production of NUCB2/nesfatin-1 by the stomach and adipose tissue fluctuates with age to regulate energy homeostasis during different states of development.
