Endocrine Abstracts

Visceral fat (VAT) may represent the more important site of endocannabinoid dysregulation in obesity. No data are available on the expression of the ECs in morbid obesity. Eleven morbidly obese women, who underwent bariatric surgery, were enrolled and paired samples of VAT and subcutaneous fat tissue (SAT) were obtained from each patient. Real-Time PCR for the gene expression levels of cannabinoid type 1 (CB1) receptor, N-acyl-phosphatidil-ethanolamine (NAPE), and fatty acid amide hydrolase (FAAH) which respectively sinthesize and degradate anandamide (AEA) and, diacylglycerol lipase (DAGL-β) and monoacylglycerol lipase (MAGL), which respectively synthesize and degrade 2-arachidonoyl-glycerol (2-AG) was performed. After extraction and after exclusion of genomic DNA contamination, 1 μg RNA was reverse transcribed using oligodT primers. Real-time cDNA quantification was performed by a thermocycler. iCycler iQ^® (BioRad). CB1, FAAH, NAPE-PLD, DAGL-β and MAGL primers for SYBR^® Green analysis were designed by ‘Beacon Designer^®’ and synthesized by Invitrogen. Assays were performed in duplicate and a standard curve from consecutive 10-fold dilutions of a cDNA pool representative of all samples, was included for each determination. Relative expression analysis was corrected for PCR efficiency and normalized respect to reference gene β-actin. BMI was 46.3±1.38 kg/m². Total cholesterol was 186±12.8 mg/dL, HDL-cholesterol 53.3±3.74 mg/dL and triglycerides 118±16.3 mg/dL. CB1 mRNA was significantly higher in SAT than in VAT. VAT had a significantly higher expression of NAPE than SAT. There was no difference in FAAH mRNA between SAT and VAT. SAT displays a higher expression of MAGL than VAT, not reaching the statistical significance. DAGL-β was more significantly expressed in SAT than in VAT. These data, obtained in morbid obesity, suggest that the ECs plays a crucial role not only in VAT but also in SAT, underlying the differences in the ECs dysregulation when morbid obesity without metabolic alterations is compared to abdominal obesity with metabolic alterations.