title: Proteomic analysis of mature adipocytes from obese patients in relation to aging.
abstract: Obesity and aging are interrelated conditions that both cause changes in adipocyte metabolism and affect the distribution of fat in both subcutaneous and visceral depots. In addition, both weight gain and aging can lead to similar clinical outcomes such as insulin resistance, cardiovascular disease, type 2 diabetes mellitus, atherosclerosis and stroke. Our objective was to examine the changes in protein expression within the subcutaneous adipose tissue of obese patients, matched for BMI, in relation to age. Mature adipocytes were isolated from liposuction samples of abdominal subcutaneous adipose tissue collected from both young (26.2±4.3 (mean age±SD); n=7) and old (52.2±4.7 (mean age±SD); n=7) obese individuals. Total protein extracts were then compared by two-dimensional difference in gel electrophoresis (2D DIGE). Thirty differentially expressed protein spots (ANOVA test, p≤0.05; fold-change ≥1.8) were detected, of which, 15 were identified by MALDI-TOF mass spectrometry. These were comprised of a total of thirteen unique protein sequences. Nine proteins were more abundant in the adipocytes isolated from old vs. young individuals. These proteins included prohibitin 1, protein disulphide isomerase A3, beta actin, profilin, aldo-ketoreductase 1 C2, alpha crystallin B and the annexins A1, A5 and A6. Four other proteins were less abundant in the adipocytes from old, obese subjects and these included keratin type 2 cytoskeletal 1, keratin type 2 cytoskeletal 10 and hemoglobins A and B. The differentially abundant proteins were investigated by Ingenuity Pathway Analysis (IPA) to reveal their associations with known biological functions. This analysis identified signal transducer and activator of transcription 3 as the central molecule in the connectivity map and the apoptotic pathway as the pathway with the highest score. Differences in the abundances of several proteins were confirmed by immunoblotting: i.e., prohibitin 1, protein disulphide isomerase A3, beta actin, profilin and signal transducer and activator of transcription 3 proteins. In conclusion, proteomic analysis of subcutaneous adipose tissue reveals differences in the abundance of proteins in adipocytes isolated from young vs. old individuals. These differentially abundant proteins are involved in the regulation of apoptosis, cellular senescence and inflammatory response. All these are common pathologic events in both obesity and aging.
authors.affiliation: Obesity Research Center, College of Medicine, King Saud University, P.O. Box 2925 (98), Riyadh 11461, Saudi Arabia; Department of Medicine, College of Medicine, King Saud University, P.O. Box 2925 (38), Riyadh 11461, Saudi Arabia. Electronic address: aalfadda@ksu.edu.sa.
authors.forename: Assim A
authors.initials: AA
authors.lastname: Alfadda
authors.forename: Hicham
authors.initials: H
authors.lastname: Benabdelkamel
authors.forename: Afshan
authors.initials: A
authors.lastname: Masood
authors.forename: Amr
authors.initials: A
authors.lastname: Moustafa
authors.forename: Reem
authors.initials: R
authors.lastname: Sallam
authors.forename: Abdulelah
authors.initials: A
authors.lastname: Bassas
authors.forename: Mark
authors.initials: M
authors.lastname: Duncan
mesh_terms: D000199:Actins / Q000378:metabolism; D017667:Adipocytes / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D000375:Aging / Q000378:metabolism* / Q000473:pathology; D017209:Apoptosis; D005260:Female; D006801:Humans; D008875:Middle Aged; D009765:Obesity / Q000378:metabolism* / Q000473:pathology; D000091402:Prohibitins; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D060066:Protein Interaction Maps; D040901:Proteomics; D012097:Repressor Proteins / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D050151:Subcutaneous Fat / Q000378:metabolism / Q000473:pathology; D055815:Young Adult
chemical_list: D000199:Actins; C000717328:PHB protein, human; D000091402:Prohibitins; D012097:Repressor Proteins; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human
keywords: 2 dimensional difference in-gel electrophoresis; 2D-DIGE; ACTB; AKR1C2; Aging; BMI; CRYAB; CVD; ECM; ER; FFA; Hb; Human adipocytes; IEF; IPA; LDL; MALDI; MS; O; Obesity; PDIA3; PHB1; Prohibitin1; Proteomics; SCAT; SD; STAT3; SVF; TOF; Tg; UPR; VAT; Y; aldo-keto reductase C2; alpha crystallin b; beta actin; body mass index; cardiovascular disease; endoplasmic reticulum; extracellular matrix; free fatty acids; hemoglobin; ingenuity pathway analysis; isoelectric focusing; low-density lipoprotein; mass spectrometry; matrix-assisted laser desorption/ionization; old; prohibitin 1; protein disulphide isomerase A3; signal transducer and activator of transcription 3; standard deviation; stromo vascular fraction; subcutaneous adipose tissue; time-of-flight; triglycerides; unfolded protein response; visceral adipose tissue; young