ADIPOSE TISSUE METABOLISM

BCH 303 (E. C. UDEGBUNAM)

Adipose tissue known as body fat is a connective tissue that extends throughout the body. It is
found under the skin (subcutaneous fat), between the internal organs (visceral fat, that surrounds
the abdominal cavities) and inner cavities of bones (bone marrow adipose tissues).
Adipose tissue is classified into two types
1. White adipose tissue (WAT)
2. Brown adipose tissue (BAT)
White adipose tissue is critical for
1. Insulin sensitivities
2. Energy storage
3. Endocrine communication

BAT (present in mammals postnatally, and during hibernation) used for non-shivering heat
production and is critical for maintenance of body temperature. Brown and white adipocytes
differ in shape, size, and intracellular structure of their organelles. WAT are generally spherical
and contains single lipid droplets that pushes other organelles to its periphery. Brown adipocytes
contains multiple lipid droplets dispersed in an ellipsoidal cell rich in iron containing
mitochondria that gives the cell a brownish hue.

Adipose tissue stores body fats

Most lipid fat fall into the category of fatty acids, triacylglycerols, glycerophospholipds,
sphingolipds, eicosanoids, cholesterol, steroid hormones and fat soluble vitamins. They have
diverse chemical structure but have one common characteristic, their relative insolubilities in
water. They have diverse functions but in the context of energy and fuel storage, fatty acids
stored as triacylglycerol serve as fuels providing the body with its major source of energy.
Fatty acids are stored as esters of glycerol. Most fatty acids in humans are stored as TAG
(Triacylglycerols). In Triacylglycerol (TAG), three (3) different or same fatty acids are esterified
to a glycerol backbone. Mono and Diacylglycerol can occur during metabolism either during
degradation in synthesis.
Hydrophobicity of TAG and energy storage
TAGs are insoluble in water because the long chain fatty acids cannot form hydrogen bonds with
the water molecules and they tend to associate with themselves. This chemical characteristics has
a direct consequence on the storage capacities of TAG and assembly of biological membranes.
They are more efficient stores than glycogen. TAG yields about 2.5 times ATP when compared to
glycogen by weight. (1g of TAG will yield more ATP compared to 1g of fat) in complete
oxidation. Energy yield per gram of TAG is about four times compared with hydrated glycogen.
The average 70kg person is about 350g of glycogen in liver and muscle. This individual can store
as high as approximately 10kg of TAG. Unlike glycogen stores which are limited, TAG stores
can expand. Even though TAGs are excellent fuel store, it can easily result to obesity and
diabetes.

Transport of fatty acids and primary products

Transport and storage of fatty acids is regulated by dietary status. The fate of TAG differs in the
fed state and fasting. In the fed state, TAG is stored with net deposition in the adipose tissue.
During fasting, TAG is hydrolysed and the products are distributed around the body for energy
production. In prolonged fasting, liver converts fatty acids (product of TAG hydrolysis) to ketone
bodies, acetoacetate and β-hydroxybutyrate.

Lipid transport in the fed state

Lipids are digested in the stomach and small intestine by gastric and pancreatic lipase, with
principal products being 2-monoacyl glycerol and free fatty acids

The epithelial cells absorbs this products and are reesterified into TAG still in the epithelial cells
and packaged into chylomicrons, a triacylglycerol lipoprotein. Before esterification occurs, Fatty
acids are activated before esterification.

N.B: TAG synthesis is different between intestinal cells and other tissues. 2-MAG is and
intermediate in intestinal tissues while phosphatidic acid is the intermediate in other tissues.
Triacylglycerol is hydrophobic and so would coalesce when transported alone thereby impeding
blood flow. TAG are packaged in lipoprotein particles called chylomicrons. Other components of
the chylomicrons are cholesterol, cholesteryl esters, fat soluble vitamins. The protein constituent
of the lipoproteins are called apoproteins, with the major apoprotein of chylomicron being apo-
B48 as they leave the intestinal cells. Apo-B48 is similar to the major apoprotein of VLDL
synthesised in the liver. Both Apo-B48 and Apo-B100 are encoded by a single gene.

Chylomicrons are secreted into the lymph by exocytosis and then circulate in the blood stream.
The chylomicrons released into the blood stream at this stage are nascent chylomicrons. As the
accept proteins from HDL they become mature chylomicrons. HDL transfers aop-E and apo-CII
to chylomicrons. Apo-E plays a role in receptor recognition for receptors on liver cells while
apo-CII acts as an activiator of LPL. Apart from the diet, the liver is another source of TAG in
the fed state. Fatty acids are synthesised in this tissue (liver) from excess carbohydrates and
amino acids. Fatty acids synthesised in the liver and assembled in to TAG are packaged into very
low density lipoprotein (VLDL) which is secreted into the blood stream. TAG in chylomicrons
and VLDL are hydrolysed by lipoprotein lipase expressed by certain tissues such as the adipose
tissue and this is facilitated by apoprotein-cII, present in chylomicrons and VLDL. Adipose
tissue in turn take up the products (fatty acids and mono acylglycerol) and is assembled into
TAG, allowing net deposition of fuel in the adipose tissue.

Chylomicrons Proportion by weight of components of

chylomicrons
Lipid transport in the fasting state
In the fasted state, the TAG stored in the adipose tissue are mobilised for used as fuel. This is
catalysed by a hormone sensitive lipase activated when phosphorylated by cAMP dependent
protein kinase A. Insulin prevents this phosphorylation. Another protein perilipin plays a role and
its activity is determined by its phosphorylated state. When phosphorylated, hormone sensitive
lipase becomes active. When dephosphorylated, hormone sensitive lipase remains inactive
because its translocation to the surface of fat droplet is blocked.

Adipose tissue in regulation of lipid metabolism

Adipose tissue store body fat as neutral TAG and represent the chief energy reservoir within
mammals. Adipose tissue store TAG under conditions of surplus and release fatty acid to supply
other tissues during fasting or times of high energy demand. Adipose tissue is central to
regulation of systemic lipid metabolism and nutritional and hormonal factors balance lipid
storage and breakdown within the fat cell.
As mentioned previously, one of the major roles of adipose tissue (adipose) to maintain a critical
balance between lipogenesis and lipolysis in order to maintain whole body insulin sensitivity and
energy homeostasis.

Lipogenesis (synthesis of fatty acids from non-lipid conversion of glycerol)

Adipocytes accumulate lipid via one of two factors
1. Under normal feeding conditions
2. Denovo lipogenesis

1. Normal feeding conditions: adipose tissue take up dietary lipids from circulation in the
form of free fatty acids (FFAs), liberated from circulating TAG via the action of
lipoprotein lipase (LPL). The LPL is transported to adjacent capillary lumen to catalyse
the release of FFA from TAG-containing lipoprotein which can be chylomicrons (the
lipoprotein that binds dietary TAG) or VLDL (the lipoprotein that binds TAG synthesized
by liver).
The reactions of lipogenesis also involve uptake of glucose via GLUT4 and the subsequent
conversion of glycerol which serves as backbone for sequential esterification of fatty acids to
form TAG.
 2. De novo lipogenesis: a high carbohydrate meal implies and excess glucose oxidation
yielding elevated levels of acetyl coA that becomes substrate to generate fatty acids. This
occurs through the enzyme acetyl coA carboxylase (ACC1) and fatty acid synthase (FAS)
to convert acetyl coA to palmitate.

Lipolysis
Under physiological conditions when metabolic fuels are low such as during fasting, extreme
exercise and cold exposure. Adipocytes mobilise their TAG by breaking down TAG to supply
fuel to peripheral tissues

This process occurs in all tissues but most prevalent in adipose tissues where the bulk of the TAG
is stored. There is a sequential activity of the adipocyte triglyceride lipase (ATGL), hormone
sensitive lipase (HSL) and monoacylglycerol lipase (MGL) whose action release glycerol which
can be reesterified within the adipocytes or released into circulation to be used by other tissues.
Hormonal regulation in adipose tissues
In addition to strong TAG adipocytes secrete hormones that regulate glucose and fat metabolism.
The hormones are leptin, adiponectin and resistin. They are all secreted by adipocytes under
different conditions. Their roles is best understood in mouse models.
Leptin is released from adipocytes as TAG levels increase. Leptin signals the hypothalamus to
reduce eating and increased physical activity. Mice lacking the hormone leptin are obese.
Injection of leptin causes them to lose weight. Leptin is secreted by adipcoytes along with other
tissues to act on hypothalamic leptin receptors (Ob-Rb) to decrease food intake and increase
energy expenditure in the host. Under normal condition, the amount of leptin produced by fat
tissues is proportional to the mass of adipose tissues. Leptin deficiency or resistance lead to
hyperphagia and decreased energy expenditure. This predictably, leads to insulin resistance,
diabetes mellitus and decrease in lean body mass. (see powerpoint)
Resistin is associated with insulin resistance. Type two diabetes drug known as thiaolidinedione
act by suppressing the transcription of resistin.
Adiponectin is secreted from adipocyte in inverse proportion to their adipose mass. Lean people
secrete more adioponectin than obese individual. Adiponectin enhances fatty acid oxidation
through activation of AMP-activated kinase, a cellular energy sensor which inhibits acetyl
carboxylase, a rate limiting enzyme in de novo lipogenesis. The result is malonyl coA production
is reduced and enhances fatty acid oxidation. In addition to its insulin sensitising and glucose
lowering effects in the liver and skeletal muscle, adiponectin is cardioprotective. Low circulating
adiponectin levels correlate significantly with coronary artery disease.

Role of insulin in adipose tissue metabolism

After meal, there is increase in insulin levels that suppress lipolysis by increasing the activity of
phosphodiesterase 3(PDE3B) and increasing cAMP levels. During fast, insulin levels drop and
noradrenalin is released promoting lipolysis. Exercise increasing the levels of noradrenalin,
growth hormone, cortisol with increased exercise intensity while insulin levels reduce.
When adipose tissue become resistant as seen in patient with diabetes and sometimes in obesity,
insulin ability to inhibit lipolysis is impaired. This condition causes an excessive lipolysis
leading to excess FFA in both fasted and fed state. Constant exposure of liver and muscle to these
high FFA promotes uptake and ectopic storage of lipids in these tissues. Ectopic lipids impair
insulin signalling and this insulin resistance at the level of adipocyte via increased lipolysis. (see
powerpoint)

Mechanism of action of insulin

Intracellular signaling of insulin is mediated by Insulin Receptor Tyrosine Kinase (IRTK). There
are various insulin signaling pathway but with regards to glucose metabolism, the binding of
Insulin to IRTK induces a conformational change where certain tyrosine residues on IRTK are
autophosphorylated. This is followed by activation of phosphotyrosine binding protein such as
Insulin Receptor Substrate (IRS). IRS activation is by phosphorylation which then recruits
phosphatidylinositol-3-OH kinase (PI3K) and catalyzes the production of phosphatidylinositol-
3,4,5-trisphosphate (PIP3) from phosphatidylinositol-4,5-bisphosphate (PIP2). PIP3 recruits Akt
at the membrane and is activated by 3-phosphoinositide-dependent kinase-1 (PDK1) and
mechanistic target of rapamycin complex 2 (mTORC2). Akt subsequently phosphorylates
downstream, substrates in muscle, liver and adipose tissue which elicit insulin-induced nutrient
reservation in these tissues.

Insulin resistance
Insulin resistance is physiologically defined as an inability of some type of tissues to respond to
normal insulin levels, and thus, higher than normal levels of insulin are required to maintain the
normal functions of insulin. In the fasted state, liver secretes glucose into the blood. This is
called: Hepatic Glucose Production (HGP). The process of HGP involves Hepatic Glycogen
breakdown or denovo glucose synthesis through gluconeogenesis.
After food intake, insulin secreted by pancreatic β-cells promotes anabolism and suppresses
catabolic programs. During glucose metabolism, insulin stimulates several glucose-consuming
tissues, such as skeletal muscle and adipose tissues, to uptake glucose and then promotes the
syntheses of glycogen and lipid in liver, skeletal muscle, and adipose tissue. In addition, insulin
suppresses HGP by inhibiting the expressions of gluconeogenic genes and lipolysis in adipose
tissue.
The primary function of hepatic insulin signaling is to decrease HGP by repressing
gluconeogenesis mediated by the Akt-induced phosphorylation of forkhead box O1 (FOXO1)
which excludes FOXO1 from the nucleus, and thus, prevents the transcriptional activations of
gluconeogenic gene expressions, such as glucose-6-phosphatase (G6PC) and
phosphoenolpyruvate carboxylase (PEPCK).

Insulin resistance in liver and adipose tissue

The liver critically controls postprandial carbohydrate levels by suppressing HGP and
stimulating the deposition of glucose as glycogen and is the primary source of glucose
production during fasting. In T2DM patients, insulin cannot regulate hepatic glycogen synthesis
or glucose production, and increased hepatic gluconeogenesis is the primary cause of fasting
hyperglycemia in T2DM. Defective suppression of hepatic gluconeogenesis in insulin resistance
is largely associated with lipolysis defects in adipose tissue and the de-suppression of FOXO1
transcription factor in liver.

Mechanism of Insulin Resistance

Glucose-fatty acid cycle (Randle cycle)
It was asserted that increased fatty acid oxidation in the obese impaired in the use of glucose as a
source of fuel by inhibiting the activities of key glycolytic enzymes. According to Randle et al.,
fatty acid oxidation might increase mitochondrial acetyl-CoA levels through β-oxidation and
subsequently inactivate pyruvate dehydrogenase, which in turn, would increase intracellular
citrate levels and inhibit phosphofructokinase (a key glycolytic enzyme), and lead to the
accumulation of intramyocellular glucose-6-phosphate, which inhibits hexokinase activity and
causes the accumulation of intramyocellular glucose.

Ectopic Lipid Accumulation

Ectopic fat accumulation in peripheral tissues such as the liver and skeletal muscle can lead to
severe IR. Several lipid metabolites such as diacylglycerol, lysophosphatidic acid, ceramides,
and acylcarnitine are involved in the ectopic lipid accumulation-induced IR in the liver and
skeletal muscle. The accumulation of lipids is a as a result of impaired fatty acid oxidation. This
impairment leads to the redirection of long-chain acyl CoA molecules into endoplasmic
reticulum (ER)-localized and cytosolic lipid species, such as diacylglycerols, ceramides, and
triglycerides. In the liver and skeletal muscle, elevated levels of diacylglycerol trigger the
movement of nuclear protein kinase C (PKC) to the plasma membrane. This, in turn, hinders
IRTK activity by phosphorylating Thr1160, ultimately leading to the deactivation of IRS2, PI3K,
and AKT2.

Inflammation in Insulin resistance

In obesity, there is an increase in fat accumulation in the adipose tissue,
leading to larger adipocyte sizes, expansion of adipose tissue, and changes
in pro-inflammatory cytokines. Obesity leads to increase infiltration of
macrophages which increases cytokine production. The inflammatory
response can proceed in two ways: Stimulation of cytokines leads to the
phosphorylation of serine in IRS1, thereby inducing IR. Secondly
inflammation, alters lipid metabolism with TNF-α inducing lipolysis and altering the
production of IL-6. Studies have shown that knockdown of TNF-α reduces IR.