Hormone-sensitive lipase serine phosphorylation and glycerol exchange across skeletal muscle in lean and obese subjects: effect of beta-adrenergic stimulation

Johan W E Jocken, Carsten Roepstorff, Gijs H Goossens, Paula van der Baan, Marleen van Baak, Wim H M Saris, Bente Kiens, Ellen E Blaak, Johan W E Jocken, Carsten Roepstorff, Gijs H Goossens, Paula van der Baan, Marleen van Baak, Wim H M Saris, Bente Kiens, Ellen E Blaak

Abstract

Objective: Increased intramuscular triacylglycerol (IMTG) storage is a characteristic of the obese insulin-resistant state. We aimed to investigate whether a blunted fasting or beta-adrenergically mediated lipolysis contributes to this increased IMTG storage in obesity.

Research design and methods: Forearm skeletal muscle lipolysis was investigated in 13 lean and 10 obese men using [(2)H(5)]glycerol combined with the measurement of arteriovenous differences before and during beta-adrenergic stimulation using the nonselective beta-agonist isoprenaline (ISO). Muscle biopsies were taken from the vastus lateralis muscle before and during ISO to investigate hormone-sensitive lipase (HSL) protein expression and serine phosphorylation.

Results: Baseline total glycerol release across the forearm was significantly blunted in obese compared with lean subjects (P = 0.045). This was accompanied by lower HSL protein expression (P = 0.004), HSL phosphorylation on PKA sites Ser(563) (P = 0.041) and Ser(659) (P = 0.09), and HSL phosphorylation on the AMPK site Ser(565) (P = 0.007), suggesting a blunted skeletal muscle lipolysis in obesity. Total forearm glycerol uptake during baseline did not differ significantly between groups, whereas higher net fatty acid uptake across the forearm was observed in the obese (P = 0.064). ISO induced an increase in total glycerol release from skeletal muscle, which was not significantly different between groups. Interestingly, this was accompanied by an increase in HSL Ser(659) phosphorylation in obese subjects during ISO compared with baseline (P = 0.008).

Conclusions: Obesity is accompanied by impaired fasting glycerol release, lower HSL protein expression, and serine phosphorylation. It remains to be determined whether this is a primary factor or an adaptation to the obese insulin-resistant state.

Figures

FIG. 1.
FIG. 1.
Plasma glycerol TTR during 6-h primed constant infusion of [2H5]glycerol (n = 3) in arterialized blood (▪), forearm venous blood (•), and expected forearm venous enrichment (○). The expected deep venous glycerol enrichment was calculated as arterialized enrichment multiplied by arterialized glycerol concentration divided by deep venous glycerol concentration. The measured venous enrichment was consistently lower than the expected deep venous enrichment (P < 0.05), implying uptake of glycerol across the forearm. Values are means ± SE.
FIG. 2.
FIG. 2.
Total glycerol uptake (A) and release (B) across the forearm during baseline (▪) and ISO infusion (□) using a [2H5]glycerol tracer in lean and obese subjects. *P < 0.05 obese vs. lean; #P < 0.05 ISO vs. baseline. Values are means ± SE.
FIG. 3.
FIG. 3.
HSL protein expression (A) and Ser563 (B), Ser565 (C), and Ser659 (D) phosphorylation during baseline (▪) and ISO infusion (□) in lean and obese subjects. Data are expressed as arbitrary units (AU). *P < 0.05 obese vs. lean; †P < 0.01 obese vs. lean in change between baseline and ISO. Values are means ± SE.

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