140 Relaxin Regulates Porcine Adipose Tissue Development by Inhibiting Preadipocyte Number, Stimulating Lipolysis, and Upregulating mRNA Expression of Adipokine, Fatty Acid Metabolism and Extracellular Matrix Genes

Abstract

Relaxin (RLN) is best known as a reproductive hormone that remodels cervical connective tissue. However, porcine adipose tissue can secrete and respond to RLN, suggesting locally derived RLN could modulate adipose tissue development and function. To test this hypothesis, primary cultures of porcine subcutaneous adipose tissue stromal-vascular (AT-SV) cells and visceral adipose tissue explants were utilized as model systems to determine the effect of 100 ng/ml exogenous RLN on 1) preadipocyte proliferation and differentiation, 2) the mRNA expression of adipokine, extracellular matrix protein and fatty acid metabolism genes, and 3) lipolytic rate in porcine adipose tissue. AT-SV cells were harvested from 3-day-old neonatal pigs (n=6) and plated in vitro in growth medium containing either vehicle or RLN for cell number assays or were induced to differentiate by treatment with insulin, hydrocortisone, and rosiglitazone for 8 days in the presence or absence of RLN for adipogenesis assays. Total RNA was extracted from differentiating cultures on days 0 and 8 post-induction. Real-time PCR was then utilized to determine changes in mRNA expression for target genes. For lipolysis and cell signaling experiments, visceral adipose tissue was obtained from market weight pigs at harvest, minced into explants, and cultured in serum-free medium in the presence or absence of RLN and cAMP pathway modulators with differences in glycerol release determined following 2-hour treatment. Overall, RLN decreased preadipocyte number 1.27-fold under serum-free conditions (P < 0.05), enhanced adipogenesis based upon 2.4-fold increase in GAPDH activity (P < 0.05) and a 2.2-fold increase in ORO accumulation (P < 0.05) while significantly increasing the mRNA expression of multiple metabolic, adipokine, and extracellular matrix component genes. RLN treatment stimulated glycerol release by 3-fold via the cAMP pathway (P < 0.01). Collectively, these data support the hypothesis that RLN regulates adipose tissue development through stimulating adipogenesis and modulating adipocyte metabolism.