1 .

ENERGY HOMEOSTASI S
2 . GPRC RECEPTORS
3 . SCFAS
4. I NSULI N SI GNAI NG
Background
Conceptual Flow
Keywords
Adipocytes
Abbreviations
MA: mature adipocytes (PPAR): peroxisomes proliferator-
activated receptor
GLP1: glucagon-like peptide aP2: Adipocyte fatty acid binding
protein
Background: Adipocytes
Adipocytes

Adipocyte differentiation
White adipose tissue (WAT) is the major energy
reserve in higher eukaryotes, and storing triacylglycerol
in periods of energy excess and its mobilization during
energy deprivation are its primary purposes
Mature adipocytes, the main cellular component of
WAT, are uniquely equipped to function in energy
storage and balance under tight hormonal control.

PPAR is the obligatory transcription factor
indispensable for the differentiation and survival of
both white and brown adipocytes
Background: WAT and energy homeostasis
Fasting State Post-prandial
http://www.nature.com/cr/journal/v17/n6/full/cr200748a.html#fig2
Background: Insulin the anabolic hormone
Insulin stimulates glucose uptake in the
liver, fat cells and skeletal
muscle. Glucose is stored as either
glycogen in liver and muscle, or as
triglycerides in adipocytes.
Glucose is stored as either glycogen in
liver and muscle, or as triglycerides in
adipocytes.
Important actions of insulin
include inhibition of lipolysis,
glycogenolysis, and
gluconeogenesis.
http://www.medbio.info/horn/time%203-4/how_insulin_works.htm
Background: The role of SCFAs in energy homeostasis
de novo synthesis
of lipids, glucose
signaling
molecules



G protein-
coupled
receptors
GPR41
GPR43
GPR43

Regulation of inflammatory
responses
GPR43 promotes leptin secretion,
adipogenesis and inhibition of
lipolysis in adipose tissue and
adipocytes

Bacterial fermentation of indigestible prebiotics
GPR43-
knockout
mice
exhibit
obesity
GPR43 is
abundantly
expressed
in WAT
Adipose-
specific
GPR43
transgenic
mice are
lean
GPR43
suppresses
insulin
signaling in
adipose
tissue
GPR43
promotes
energy
expenditure
GPR43
suppressess
insulin
signalling via
G(i/o)-
PLC-PKC-
PTEN
General
Methodology
1. Trasgenic mice:

Gpr43 is abundantly expressed in the WATs
Gpr43 was most abundantly expressed in the WAT
and was not expressed in the BAT.

During adipogenesis, Gpr43 was expressed in the
later stages of differentiation than aP2 and PPARg.
3T3-L1 and 3T3-F442A mouse cell lines
differentiate spontaneously when
exposed to a hormonal cocktail
I. Gpr43 is abundantly expressed in the WATs
GPR43 was expressed in the later stages of
differentiation than aP2 and PPAR
http://www.hindawi.com/journals/ppar/2008/679137/fig1/
aP2 is a key mediator of intracellular transport and metabolism of
fatty acids. Its expression during adipocyte differentiation is regulated
through the actions of PPAR and CCAAT/enhancer binding protein
alpha (C/EBPalpha).
PPAR induces the expression of genes involved in lipid
synthesis and storage through enhancers activated during
adipocyte differentiation.
Gpr43 is abundantly expressed in the WATs
Stromal vascular fraction (VSF) - preadipocytes, fibroblasts,
vascular endothelial cells and a variety of immune cells (i.e. adipose tissue
macrophages (ATMs))
Gpr43 knockout mice are obese
P1 WATand BAT
were stained with
oil red P1 Body weight P1 fat mass
Protein
expression
Expression of aP2 and Pparg
mRNA
Haematoxylineosin stained
WAT and mean area of
adipocytes
Gpr43 knockout mice are obese
Insulin tolerance
test
Glucose tolerance
test
Euglycaemic
hyperinsulinaemic clamp in
HFD-fed mice
Glucose
infusion
ratio
Glucose
disposal
rate
Hepatic
glucose
production
suppression



Hyperinsulinemic Euglycemic Glucose Clamp Technique. After an overnight fast,
insulin is infused intravenously at a constant rate that may range from 5~120
mU/m
2
/min (dose per body surface area per minute). This constant insulin infusion
results in a new steady-state insulin level that is above the fasting level
(hyperinsulinemic). As a consequence, glucose disposal in skeletal muscle and
adipose tissue is increased while hepatic glucose production is suppressed. Under
these conditions, a bedside glucose analyzer is used to frequently monitor blood
glucose levels at 5~10 min intervals while 20% dextrose is given intravenously at a
variable rate in order to "clamp" blood glucose concentrations in the normal range
(euglycemic). An infusion of potassium phosphate is also given to prevent
hypokalemia resulting from hyperinsulinemia and increased glucose disposal. After
several hours of constant insulin infusion, steady-state conditions can typically be
achieved for plasma insulin, blood glucose, and the glucose infusion rate (GIR).
Assuming that the hyperinsulinemic state is sufficient to completely suppress hepatic
glucose production, and since there is no net change in blood glucose concentrations
under steady-state clamp conditions, the GIR must be equal to the glucose disposal
rate (M). Thus, whole body glucose disposal at a given level of hyperinsulinemia can
be directly determined (Adopted from Muniyappa R, et al. Am J Physiol Endocrinol
Metab 294:E15-26, 2008).
Mechanisms of insulin sensitivity and
resistance in muscle and liver(A) Insulin-
sensitive muscle. (B) Insulin-resistant muscle.
(C) Insulin-sensitive liver. (D) Insulin-resistant
liver. IRS=insulin-receptor substrate.
IR=insulin receptor. PI3K=1-
phosphatidylinositol 3-kinase. GLUT4=glucose
transporter 4. DAG=diacylglycerol.
PKC=protein kinase C. Ser=serine.
Thr=threonine. FOX01=forkhead box O1.
FOXA2=forkhead box A2. G6P=glucose-6-
phosphate. GS=glycogen synthase.
GSK=glycogen synthase kinase. Green circle
with plus sign represents activation. Red circle
with minus sign represents inactivation. Solid
line with arrowhead represents increase or
accumulation of substrate. Dotted line
indicates inhibition of pathway.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2995547/pdf/nihms250988.pdf
Gpr43 knockout mice are obese
Level of
inflammation
Expression of F4/80 and TNF-
Fecal major SCFA and
plasma acetate contents in HFD-
fed mice
Comparison of
microbial
communities in
HFD-fed mice
Communities of gut microbiota
in Gpr43/ mice exhibited an
increase in firmicutes, an
SCFA- producing phylum, and a
decrease in - proteobacteria
and actinobacteria
Body weights
of HFD-fed
mice under
GF conditions
Body weight
changes under GF
to CONV conditions
Gpr43 knockout mice are obese
Insulin and glucose tolerance under
antibiotic treatment
Acetate and glucose
plasma concentrations
after 1 h of feeding (24 h
fasting)
Plasma acetate concentration
with or without antibiotic
treatment during feeding
Effect of acetate on body
weights and fat mass
under antibiotic
treatment
Adipose tissue-specific Gpr43 transgenic mice are lean
Subcutaneous
WAT & inters-
capular BAT of
P1 were stained
with oil red O P1 Body weight
P1 fat mass
Protein
expression
Expression of aP2 and Pparg
mRNA
Haematoxylineosin stained
WAT and mean area of
adipocytes
Adipose tissue-specific Gpr43 transgenic mice are lean
Comparison of
microbial
communties
Fecal & plasma major SCFA
Body weight & fat mass of aP2-
Gpr43TG mice fed an HFD
Body weight & fat mass
under antibiotic treatment
Adipose tissue-specific Gpr43 transgenic mice are lean
ITT & GTT of aP2-Gpr43TG mice fed
an HFD
Plasma glucose
concentration in
aP2-Gpr43TG mice
fed an HFD
Euglycaemic
hyperinsulinaemic clamp
aP2-Gpr43TG mice fed an
HFD
Glucose
infusion
ratio
Glucose
disposal
rate
Hepatic
glucose
production
suppression
ITT & GTT of aP2-Gpr43TG mice fed
an HFD under antibiotic treatment
Oil red O-stained liver and
hepatic triglyceride content in
aP2-Gpr43 TG mice fed an
HFD
Level of WAT inflammation
Gpr43 suppresses insulin signaling in the adipose tissues but not in muscles or liver
Insulin-stimulated Akt phosphorylation of Ser473 in the
WAT, muscles & liver
Inhibitory effects of acetate on insulin signaling
Gpr43 suppresses insulin signaling in the adipose tissues but not in muscles or liver
Effect of acetate on glucose uptake in MEF-derived
adipocytes from Gpr43-/-or aP2-Gpr43TG mice
Effect of acetate on the fatty acid uptake in MEF-derived
adipocytes from Gpr43-/- or aP2-Gpr43TG mice
Gpr43 suppresses insulin signaling in the adipose tissues but not in muscles or liver
LPL activity in the WAT and the muscles
LPL activity of Gpr43-/- mice fed an HFD under
GF conditions or aP2-Gpr43TG mice treated with
antibiotics
Gpr43 suppresses insulin signaling via G(i/o)-PLC-PKC-PTEN signaling
Inhibitory effects of GPR43
agonists and a GPR41 agonist
on insulin-induced Akt
phosphorylation
Effects of Gi/o signaling inhibition on suppression of
insulin-induced Akt phosphorylation by acetate
Effects of Gq signaling inhibition
using siRNA on suppression of
insulin-induced Akt
phosphorylation by acetate
Effects of G signaling inhibition on suppression of
insulin-induced Akt phosphorylation by acetate
Effects of GPR43 stimulation on PTEN
phosphorylation
Gpr43 suppresses insulin signaling via G(i/o)-PLC-PKC-PTEN signaling
Effects of PTEN
signalling inhibition on
suppression of
insulin-induced Akt
phosphorylation by
acetate
Effect of GPR43 agonists (10mM acetate
and 10mM PA) on insulin-induced
glucose uptake
Effect of acetate on insulin-
induced LPL activity
Schematic diagram of the mechanism for GPR43-
mediated suppression of fat accumulation.
Gpr43 promotes energy expenditure by increasing the consumption of lipids
Biochemical analysis of plasma obtained from Gpr43/mice on an HFD as well as aP2-Gpr43TG mice
Triglycerides
Free fatty acids
mRNA levels of genes involved in energy expenditure
Energy expenditure of
Gpr43/ mice on an HFD
Total activity in Gpr43/
mice fed an HFD
RER of Gpr43/ mice fed
an HFD
Gpr43 promotes energy expenditure by increasing the consumption of lipids
Energy expenditure of aP2-
Gpr43TG mice
Total activity in aP2-
Gpr43TG mice
RER of aP2-Gpr43TG mice
Schematic model of suppression of fat accumulation via Gpr43
After feeding, SCFAs, produced by microbial fermentation in the
gut, activate GPR43 in adipose tissues.

SCFA-mediated GPR43 activation suppresses insulin-mediated fat
accumulation and thereby regulates the energy balance by suppressing
accumulation of excess energy and
promoting fat consumption.
www.intechopen.com/books/alternative-medicine/investigation-on-the-mechanism-of-qi-invigoration-from-a-perspective-of-effects-of-sijunzi-
decoction
www.jci.org/articles/view/58109/figure/1
Leptin is a 16-kDa protein hormone, which is secreted
by adipocytes
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Representative H&E stained
sections of (F) brown (BAT)
and (G) white (WAT) adipose
tissues from aP2-SufuKO and
control littermate mice.
(A and B) Microscopic view of
Oil Red O-stained cell cultures