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l-Arginine increases AMPK phosphorylation and the oxidation of energy substrates in hepatocytes, skeletal muscle cells, and adipocytes

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Abstract

Previous work has shown that dietary l-arginine (Arg) supplementation reduced white fat mass in obese rats. The present study was conducted with cell models to define direct effects of Arg on energy-substrate oxidation in hepatocytes, skeletal muscle cells, and adipocytes. BNL CL.2 mouse hepatocytes, C2C12 mouse myotubes, and 3T3-L1 mouse adipocytes were treated with different extracellular concentrations of Arg (0, 15, 50, 100 and 400 µM) or 400 µM Arg + 0.5 mM NG-nitro-l-arginine methyl ester (L-NAME; an NOS inhibitor) for 48 h. Increasing Arg concentrations in culture medium dose-dependently enhanced (P < 0.05) the oxidation of glucose and oleic acid to CO2 in all three cell types, lactate release from C2C12 cells, and the incorporation of oleic acid into esterified lipids in BNL CL.2 and 3T3-L1 cells. Arg at 400 µM also stimulated (P < 0.05) the phosphorylation of AMP-activated protein kinase (AMPK) in all three cell types and increased (P < 0.05) NO production in C2C12 and BNL CL.2 cells. The inhibition of NOS by L-NAME moderately reduced (P < 0.05) glucose and oleic acid oxidation, lactate release, and the phosphorylation of AMPK and acetyl-CoA carboxylase (ACC) in BNL CL.2 cells, but had no effect (P > 0.05) on these variables in C2C12 or 3T3-L1 cells. Collectively, these results indicate that Arg increased AMPK activity and energy-substrate oxidation in BNL CL.2, C2C12, and 3T3-L1 cells through both NO-dependent and NO-independent mechanisms.

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Abbreviations

ACC:

Acetyl-CoA carboxylase

AMPK:

AMP-activated protein kinase

AU:

Arbitrary unit

CPT-I:

Carnitine palmitoyltransferase I

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

KHB:

Krebs–Henseleit bicarbonate

MCD:

Malonyl-CoA decarboxylase

L-NAME:

NG-Nitro-l-arginine methyl ester

NO:

Nitric oxide

NOS:

Nitric oxide synthase

SCD-1:

Stearoyl-CoA desaturase 1

SDS:

Sodium dodecyl sulfate

S6K1:

Ribosomal protein S6 kinase beta-1

TTBS:

Tris–Tween-buffered saline

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Acknowledgements

We thank Dr. Stephen B. Smith and Dr. Cynthia J. Meininger for helpful discussions, as well as Mr. Scott Jobgen for assistance in laboratory analyses of metabolites. This research was supported by grants from American Heart Association–Texas (#0755024Y and 10GRNT4480020).

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  1. Department of Animal Science and Faculty of Nutrition, Texas A&M University, College Station, TX, 77843, USA

    Wenjuan S. Jobgen & Guoyao Wu

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  1. Wenjuan S. Jobgen
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Correspondence to Guoyao Wu.

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Jobgen, W.S., Wu, G. l-Arginine increases AMPK phosphorylation and the oxidation of energy substrates in hepatocytes, skeletal muscle cells, and adipocytes. Amino Acids 54, 1553–1568 (2022). https://doi.org/10.1007/s00726-022-03195-9

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