Insulin and rosiglitazone regulation of lipolysis and lipogenesis in human adipose tissue in vitro.
Lipolysis is an important process determining fuel metabolism, and insulin regulates this process in adipose tissue. The aim of this study was to investigate the long-term effects of insulin, an insulin enhancer (rosiglitazone [RSG]), and insulin in combination with RSG on the regulation of lipolysis and lipogenesis in human abdominal subcutaneous fat. Lipolysis and lipogenesis were assessed by protein expression studies of hormone-sensitive lipase (HSL) (84 kDa) and lipoprotein lipase (LPL) (56 kDa), respectively. In addition, lipolytic rate was assessed by glycerol release assay and tumor necrosis factor (TNF)-alpha release measured by enzyme-linked immunosorbent assay (n = 12). In subcutaneous adipocytes, increasing insulin doses stimulated LPL expression, with maximal stimulation at 100 nmol/l insulin (control, 1.0 +/- 0.0 [mean +/- SE, protein expression relative to control]; 1 nmol/l insulin, 0.87 +/- 0.13; 100 nmol/l insulin, 1.68 +/- 0.19; P < 0.001). In contrast, insulin at the 100 nmol/l dose reduced the expression of HSL (100 nmol/l insulin, 0.49 +/- 0.05; P < 0.05), while no significant reduction was observed at other doses. Higher doses of insulin stimulated both HSL (1,000 nmol/l insulin, 1.4 +/- 0.07; P < 0.01) and LPL (control 1.00 +/- 0.0; 1,000 nmol/l insulin, 2.66 +/- 0.27; P < 0.01) protein expression. Cotreatment with RSG induced an increased dose response to insulin for LPL and HSL (P < 0.05); RSG alone also increased LPL and HSL expression (P < 0.05). Insulin stimulated TNF-alpha secretion in a dose-dependent manner (P < 0.01); the addition of RSG (10(-8) mol/l) reduced TNF-alpha secretion (P < 0.05). In summary, chronic treatment of human adipocytes with insulin stimulates lipolysis and LPL protein expression. The addition of RSG reduced the lipolytic rate and TNF-alpha secretion. The increase in lipolysis is not explained by changes in HSL expression. These data, therefore, may explain in part why hyperinsulinemia coexists with increased circulating nonesterified free fatty acids and increased adiposity in obese and/or type 2 diabetic patients.
McTernan
Philip G
PG
Division of Medical Sciences, Department of Medicine, University of Birmingham and Heartlands Hospital, Edgbaston, Birmingham, UK. p.g.mcternan.20@bham.ac.uk
Harte
Alison L
AL
Anderson
Leah A
LA
Green
Allan
A
Smith
Stephen A
SA
Holder
Julie C
JC
Barnett
Anthony H
AH
Eggo
Margaret C
MC
Kumar
Sudhesh
S
D017667:Adipocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000328:Adult; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D005990:Glycerol / Q000378:metabolism; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology*; D008066:Lipolysis / Q000187:drug effects*; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000378:metabolism; D008875:Middle Aged; D009638:Norepinephrine / Q000494:pharmacology; D000077154:Rosiglitazone; D002787:Sterol Esterase / Q000096:biosynthesis / Q000378:metabolism; D013566:Sympathomimetics / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology*; D045162:Thiazolidinediones; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D007004:Hypoglycemic Agents; D007328:Insulin; D013566:Sympathomimetics; D013844:Thiazoles; D045162:Thiazolidinediones; D014409:Tumor Necrosis Factor-alpha; D000077154:Rosiglitazone; D002787:Sterol Esterase; D008071:Lipoprotein Lipase; D005990:Glycerol; D009638:Norepinephrine