Cannabinoid type 1 receptor blockade induces transdifferentiation towards a brown fat phenotype in white adipocytes.
OBJECTIVE
The endocannabinoid (EC) system is a major component in the control of energy homeostasis. It mediates a positive energy balance via central and peripheral pathways. Blockade of the cannabinoid type 1 receptor induces weight reduction and improves cardiovascular risk factors in overweight patients. Cannabinoid receptor type 1 (CB1R)-deficient mice are resistant to diet-induced obesity. The mechanisms responsible for these effects remain only partially elucidated. We hypothesized peripheral effects via direct modulation of adipocyte function to be an integral part of EC action on energy metabolism and insulin sensitivity.


METHODS
SV40 immortalized murine white and brown adipocytes were used for all experiments. We investigated the effect of CB1R blockade by stimulating the cells acutely and chronically with rimonabant, a selective antagonist for the CB1R, or by knocking down the receptor with small interfering RNA (siRNA). Changes in thermogenic mRNA and protein expression as well as mitochondrial biogenesis and function were assessed by real-time RT-PCR, immunoblotting, fluorescent staining techniques, electron microscopy and by measuring oxygen consumption.


RESULTS
Acute and chronic blockade of the CB1R with the selective antagonist rimonabant or by siRNA in murine white adipocytes strongly induced the thermogenic uncoupling protein-1 (UCP-1). UCP-1 expression was increased in a time- and dose-dependent manner both at the RNA and protein level. Furthermore, this effect was paralleled by enhanced peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) expression. In accordance with these findings, AMP-activated protein kinase (AMPK) phosphorylation was also increased after rimonabant treatment. Mitochondria-specific fluorescent staining demonstrated an augmentation in the number of mitochondria. This was confirmed by electron microscopy images. Moreover, rimonabant treatment enhanced the cytochrome c oxidase activity and increased cellular oxygen consumption.


CONCLUSIONS
Taken together, our data demonstrate that inhibition of peripheral CB1R action in adipocytes directly promotes transdifferentiation of white adipocytes into a mitochondria-rich, thermogenic brown fat phenotype. Enhanced thermogenesis and insulin sensitivity may represent a peripheral mechanism contributing to weight loss and improved glucose homeostasis in rimonabant-treated patients.
Perwitz
N
N
Department of Internal Medicine I, University of Luebeck, 23538 Luebeck, Germany.
Wenzel
J
J
Wagner
I
I
Büning
J
J
Drenckhan
M
M
Zarse
K
K
Ristow
M
M
Lilienthal
W
W
Lehnert
H
H
Klein
J
J
D052438:Adipocytes, White / Q000166:cytology*; D002001:Adipose Tissue, Brown / Q000187:drug effects; D000818:Animals; D063387:Cannabinoid Receptor Antagonists; D054338:Cell Transdifferentiation; D015870:Gene Expression; D051379:Mice; D009765:Obesity / Q000473:pathology*; D047495:PPAR gamma / Q000235:genetics; D010641:Phenotype; D010880:Piperidines / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D043884:Receptor, Cannabinoid, CB1 / Q000037:antagonists & inhibitors* / Q000235:genetics; D043882:Receptors, Cannabinoid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077285:Rimonabant
D063387:Cannabinoid Receptor Antagonists; D047495:PPAR gamma; D010880:Piperidines; D011720:Pyrazoles; D043884:Receptor, Cannabinoid, CB1; D043882:Receptors, Cannabinoid; D000077285:Rimonabant