LIPID METABOLISM

Romulo de Villa, MD, PhD, Cert. Biochem.

Molecular & Nutritional Oncologist
Professor of Biochemistry & Nutrition
Molecular Biology & Biotechnology Consultant
Overview
Digestive lipid metabolism
De Novo Synthesis of Fatty acid
Synthesis of Triacylglycerols
Mobilization of Stored Fats and Oxidation
of Fatty acids
Synthesis of Ketone bodies
Overview (cont.)
Phospholipid Metabolism
Glycolipid Metabolism
Metabolism of Prostaglandins and
Related compounds
Cholesterol Metabolism
Blood Lipoproteins
Emulsification of Dietary lipids in the small
intestines
- emulsification inc. the surface area
lipase activity
- detergent property of bile salt and
peristalsis
> Pancreatic activity
A. Hormonal Control of Lipid Digestion
1. CCK/pancreozymin (+)GB contraction
and release of bile; release of pancreatic
enzymes; dec. gastric motility
2. Secretin - (+) bicarbonate secretion

Digestive Lipid Metabolism
1. Triacyglycerol hydrolysis
- TAG are acted upon by pancreatic lipase
and removes FA at carbon 1 and 3
- products = 2-monoacylglycerol + FA
2. Cholesteryl ester degradation
- Cholesterol esterase
- Products: Cholesterol + FA
3. Phospholipids degradation
- phospholipase A
2

- products: lysophospholipid + FA


Absorption of Lipids by intestinal mucosal cells
- FFA, free cholesterol, 2-monoacylglycerol and
lysophospholipid together with bile salts from
mixed micelles which is absorbed at the brush
border membrane of SI
- short and medium chain FA are directly absorbed

> Resynthesis of TAG, CE and PL
2-monoacylglycerol + fatty acyl-CoA = TAG
Cholesterol + FA = CE
Lysophospholipid + FA = Phospholipid

Reverse Cholesterol Transport via HDL
Blood Peripheral
Tissues
Liver
Bile
Excess
Cholesterol
Cholesterol Catabolism into Bile Salts
Cholate Cholesterol
Cholesterol
7-hydroxylase
HO
HO
OH
COO -
OH
Bile Salts
Breakdown products of cholesterol
Amphipathic molecules
Function to transport cholesterol in the
digestive system
Structure of Biliary and Intestinal Micelles
Cholesterol
Bile Salt
Phospholipid
Functions of Micelles
Transport cholesterol from the liver into
the intestine via the biliary tree
Participate in fat digestion and absorption
Biliary Lipid Secretion
Sinusoidal Membrane
Blood Hepatocyte
Canalicular Membrane
Bile Salt
ABCG5/G8
Cholesterol
ABCB4
Phospholipid
ABCB11
Bile
Biliary Lipids
Daily Secretion (g) Lipid Class
Bile salts
Phospholipids
Cholesterol
24
11
2
Biliary Lipid Transport
Duodenum
Jejunum
Ileum
Colon
Biliary
Transport
and
Storage
Liver
Fat Digestion
Duodenum
Jejunum
Ileum
Colon
Biliary
Transport
and
Storage
Liver
F a t D i g e s t i o n
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Fatty Acids +
Lysophospholipid
Phospholipids
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Triglycerides
Fatty Acids +
Monoglycerides
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Dietary
Cholesterol
Fat Absorption
Duodenum
Jejunum
Ileum
Biliary
Transport
and
Storage
Colon
Liver
Lymph Enterocyte
Intestinal
Lumen
Cholesterol Absorption
Cholesteryl
Ester
ACAT
Cholesterol
Lymph Enterocyte
Intestinal
Lumen
Triglyceride Absorption
2 Fatty Acid
+
Monoglyceride
DGAT
Triglyceride
Lymph Enterocyte
Intestinal
Lumen
Phospholipid Absorption
Phospholipid
Fatty Acid
+
Lysophospholipid
Lymph Enterocyte
Intestinal
Lumen
Chylomicron Formation
With
apoB48
Cholesteryl
Ester
Triglyceride
Phospholipid
Acetyl-CoA Based Metabolism
De Novo Synthesis of Fatty Acids
Production of Acetyl CoA
Carboxylation of Acetyl CoA
Fatty acid synthase: a multienzyme
complex
AcetylCoA-ACP transacylase
MalonylCoA-ACP transacylase
-ketoacyl-ACO synthase
Palmitoyl thioesterase
De Novo Synthesis of Fatty Acids
Production of
Acetyl CoA

Translocation of
mitochondrial citrate
to cytosol
Conversion of citrate
to acetyl CoA +
oxaloacetate by citrate
lyase
Requirement: Inc. ATP
and Citrate

De Novo Synthesis of Fatty Acids
Carboxylation of
acetyl CoA to
malonyl CoA
Regulators of
acetylCoA
carboxylase
Activators:
insulin,Inc. CHO
intake, fat-free diet
Inhibitors: malonyl
CoA, palmitoyl
CoA,epinephrine,
fasting, high fat
diet

Carboxylation of AcetylCoA
to malonylCoA by AcetylCoA
carboxylase
Rate limiting step in fatty
acid synthesis
Coenzyme: Biotin
De Novo Synthesis of Fatty Acids
Fatty acid synthase: a
multienzyme complex
Substrate: AcetylCoA
and MalonylCoA
End Product: Palmitic
acid
Site: Cytosol
Priming Molecule: Acetyl
CoA
Rate-limiting enzyme:
Acetyl CoA carboxylase
Primary enzyme of
synthesis: Fatty acid
synthase
Carbons 15 and 16 of palmitic
acid from priming acetyl CoA
Carbons 1-14 of palmitic acid
are derived from 7 malonyl
CoA ( 2 carbons from malonyl
CoA are added 7 times to the
priming acetylCoA molecule
NADPH + H
+
are from HMP-
Shunt


1 AcetylCoA + 7 malonylCoA + 14 NADPH+ 14 H

Palmitic acid + 7 CO2 + 6H2O+ Co-A-SH +14 NADP

De Novo Fatty Acid Synthesis
1. AcetylCoA + ACP-SH AcetylCoA-ACP-transacylase Acetyl-S-ACP + CoA
2. Acetyl-S-ACP + Enzyme SH Acetyl-S-enzyme + ACP-SH
3. MalonylCoA + ACP-SH Malonyl-ACP Transacylase Malonyl-S-ACP + CoA
4. Malonyl-S-ACP + Acetyl-S-enzyme -ketoacyl-ACO synthase Acetoacetyl-S-
ACP + CO
2
5,6,7 Three steps: 2 reductions + dehydrogenation step: converts ketoacyl
group to saturated acyl group
The cycle of reactions is repeated 7X, each time incorporating a two-carbon
unit from malonyl CoA into the growing fatty acid chain

Palmitoyl-S-ACP + H
2
O Palmitoyl thioesterase Palmitate + ACP-SH




Synthesis of Triacylglycerol
Synthesis of Triacylglycerol
Intracellular Fatty acid Metabolism
Mobilization of Stored Fats: Lipolysis
Beta-Oxidation of Fatty Acids
major pathway for catabolism of saturated FA
2-carbon fragments are successively removed
from the carboxyl end of the fatty acylCoA,
producing acetylCoA
Consists of four reactions: shortening of FA by 2
carbons
Oxidation: produces FADH
2

Hydration: produces NADH
Thiolytic cleavage: produces 2 acetylCoA

ENERGY YIELD FROM -OXIDATION
From PalmitoylCoA ATP Yield
7NADH x 3 ATP by ETC oxidation 21
7 FADH
2
x 2 ATP by ETC oxidation 14
8 Acetyl CoA x 12 ATP via Krebs CAC 96
Total (Gross) 131 ATP
Less 2 ATP
NET 129 ATP
From one molecule of palmitoylCoA

Ketone Bodies: Alternative Fuel for Cells
Ketone Bodies: acetoacetic acid, BHBA,acetone
Produced in the liver when the amount of
acetylCoA exceeds the oxidative capacity of the
liver
Extrahepatic tissues that can utilize Ketone bodies
Skeletal muscles
Cardiac muscles
Renal cortex
Brain
KETOGENESIS
Utilization of Ketone Bodies
Liver produces Ketone bodies
Liver cannot use
acetoacetate as fuel ( lacks
thiophorase : enzyme for the
conversion of acetoacetate to
acetoacetylCoA
AcetoacetylCoA is converted
to 2 acetylCoA which are
oxidized by the TCA
Increased Ketogenesis
Conditions
Starvation
Severe DM
Rapid mobilization
of fat
Result to
ketonemia
ketoacidosis
Conversion of Ketone Bodies to Acetyl-CoA
Cholesterol Synthesis
Cholesterol Balance
Duodenum
Jejunum
Ileum
Colon
Portal
Venous Return
(>95% of Biliary
Secretion)
Fecal
excretion
(0.4 g/d)
Biliary
Transport
and
Storage
Liver
Synthesis
0.4 g/d
Secretion
24 g/d
Enterohepatic Circulation of Bile Salts
Duodenum
Jejunum
Ileum
Colon
Biliary
Transport
and
Storage
Fecal
excretion
(1.2 g/d)
Dietary
Cholesterol
(0.4 g/d)
Absorption
~50%
CM
apoB48
Liver
Biliary
Cholesterol
(2 g/d)
Biliary and Dietary Cholesterol
Cholesterol Balance
Duodenum
Jejunum
Ileum
Colon
Biliary
Transport
and
Storage
Cholesterol
Cholesterol
(1.2 g/d)
+
Bile Salts
(0.4 g/d)
Dietary
Cholesterol
(0.4 g/d)
Loss
(1.6 g/d)
Bile salts
Loss
(1.6 g/d)

Dietary
Cholesterol
(0.4 g/d)
Synthesis
(1.2 g/d)
Liver
Inhibitors of Cholesterol Synthesis
and Absorption
Inhibitors of Cholesterol Synthesis: Statins
Inhibit synthesis of cholesterol by cells
Lower LDL cholesterol
Mechanism: Promote LDL Clearance
LDL
Receptor
Statins
Acetate
LDL
HMG-CoA
Reductase
Cholesterol
Cholesterol Absorption Inhibitors
Inhibit absorption of dietary cholesterol
Inhibit reabsorption of biliary cholesterol
Lower LDL cholesterol
Mechanism: Inhibit LDL Formation
Plant Sterols and Stanols
Sterol/Stanol Dietary
Cholesterol
Lymph Enterocyte
Intestinal
Lumen
Plant Sterols and Stanols
Cholesterol
Cholesteryl
Ester ABCG5/G8
ACAT
NPC1L1
Lymph Enterocyte
Intestinal
Lumen
Ezetimibe
Cholesterol
NPC1L1
Cholesteryl
Ester ABCG5/G8
ACAT
Ezetimibe
X
Lymph Enterocyte
Intestinal
Lumen
Cholesterol Absorption Inhibitors
Cholesteryl
Ester
CM
apoB48
Triglyceride
Cholesterol Absorption Inhibitors
Duodenum
Jejunum
Ileum
CM
apoB48
Liver
CM Remnant
apoB48
VLDL
apoB100
Ezetimibe
X
LDL
apoB100
Colon
Dual Inhibition
Assembly and Secretion of VLDL
Presence of Triglycerides
ApoB
MTP MTP
Cholesteryl Esters
Cholesterol
Dietary/Biliary Synthesis
Effect of Cholesterol Absorption Inhibitor
Presence of Triglycerides
ApoB
Cholesterol
Dietary/Biliary Synthesis
Ezetimibe
X
MTP MTP
Cholesteryl Esters
Adding a Statin Blocks Compensatory Increase in
Synthesis
Presence of Triglycerides
ApoB
Cholesterol
Dietary/Biliary Synthesis
Ezetimibe
X X
Statin
MTP MTP
Cholesteryl Esters
Dual Inhibition
Duodenum
Jejunum
Ileum
CM
apoB48
Liver
CM Remnant
apoB48
VLDL
apoB100
Ezetimibe
X
LDL
apoB100
X
Statin
Colon
Conclusions
Cholesterol balance is regulated by both synthesis
and absorption

Inhibition of cholesterol absorption may be
compensated by increases in synthesis

Optimal LDL lowering may best be achieved by
inhibiting both pathways
The very first
symptom for 1/3
of all the heart
attacks that
occur each and
every day is . . .
INSTANT
DEATH !
Stress,
Smoking,
Lack of exercise,
Poor nutrition
and of course,
Genetics
all contribute to
HEART DISEASE!
But
the real culprit is,
HIGH
CHOLESTEROL!
High LDL-Cholesterol is a Major
Culprit for Cardiovascular Risk
Cholesterol is everywhere
in the Body ?
Inside Cells
Hepatic Tissue
Extrahepatic Tissue
Outside Cells
Connective Tissue (Blood)
Within Cell Membranes
Why is Cholesterol all over the body ?
Cells synthesize cholesterol
Cholesterol is needed
Cholesterol esterification for transfer from one
lipoprotein (HDL) to other lipoproteins (VLDL, IDL &
LDL)
Bile synthesis for emulsification of dietary fat
Steroid hormone synthesis
Androgens
Estrogens
Corticosteroid
Glucocorticoids
Cholesterol Has No Morality
There is no difference in the molecular structure
of good and bad cholesterol

The type of apoprotein holding the cholesterol will
determine the morality of cholesterol
HDL Cholesterol is Good Cholesterol
Cholesterol is brought to the liver when HDL
binds to the liver
Cholesterol is transferred to other lipoprotins
(VLDL, LDL, IDL)
Lecithin/Cholesterol AcylTransferase
(LCAT) converts cholesterol to cholesteryl
esters
Cholesterol esters transferred to other
lipoprotein (VLDL, LDL, IDL)

LDL Cholesterol is Bad Cholesterol only when
cholesterol becomes deposited on the blood vessel
wall
as part of repair of disrupted endothelial lining
due to endothelial dysfunction
if the carrier LDL is oxidized

LDL cholesterol is not all that bad
Cholesterol is brought to tissues for further
metabolism
Tissues need cholesterol
for hormone synthesis
to modulate membrane fluidity
Framingham Study
70% of men with Coronary Heart Disease (CHD) had <44
mg/dL HDL-cholesterol
1.5 % risk if HDL-c > 35 mg/dL
7.2 % risk if t-chol/HDL-c > 5
HDL-c & Trigly are low
11.5 % risk if t-chol/HDL-c > 5
HDL-c < 35 mg/dL
Trigly > 200 mg/dL
The higher the HDL-c the lower the rate of CHD
Normal Ratio of t-cholesterol / HDL-c = < 5
Low Lipid Levels in Filipinos
Triglyceride 116 mg/dL

Cholesterol 159 mg/dL
HDL-c 30 mg/dL
T-Chole/HDLc Ratio > 5
HDLc < 35
LDL-c 107 mg/dL

Evaluation: The low HDL-c is a high risk for
Filipinos
Roles of HDL Apoproteins
Brings cholesterol from peripheral tissues
(including arteries) to the liver
Removing cholesterol from arterial wall
Inhibiting growth of new plaques

Enhances stability of plaques and inhibits plaque
rupture

Provides cholesteryl esters to LDL

Protects LDL-cholesterol from oxidation by
acting as a good anti-oxidant when HDL
attaches to LDL

Reduce expression of adhesion molecules on the
vascular endothelium
Reduced adhesion of leukocytes (early phase of
atherogenesis),
prevent formation of new plaques,
maintain integrity vascular endothelium
Metabolism of
Amphipathic Lipids
Phospholipid Synthesis
Conversion of Phosphatidylethanolamine to
phosphatidylcholine
Metabolism of
Unsaturated Fatty Acids
Desaturation of Stearoyl-CoA
Why is DM a risk factor for MI?
Most type 2 diabetics die of heart attack.
Peroxisome-Proliferator activated receptor (PPAR) links
Type 2 diabetes mellitus with heart attack
PPAR activity
Free fatty acids
Free fatty acid uptake
Insulin resistance
Complement 3
Atherosclerosis is an
inflammation-based process
Why is DM a risk factor for MI?
Most type 2 diabetics die of heart attack.
Peroxisome-Proliferator activated receptor (PPAR) links
Type 2 diabetes mellitus with heart attack
PPAR activity
regulation of gene expression
Apoproteins of lipoproteins Enzymes of CHO & lipid metabolism
Free fatty acids
Free fatty acid uptake
Insulin resistance
Complement 3
Atherosclerosis is an
inflammation-based process
PPAR gamma plays a critical role in the regulation of
cholesterol homeostasis by
controlling the expression of a network of genes that
mediate
cholesterol efflux from cells
Responsible for degeneration of ABC-I, cholesterol
transporter from the cells to HDL-3
transport in plasma
Inhibits production of apolipoprotein & C3 which
destroys Apo-B (ligand that binds LDL to its
receptors) and LDL accumulates
Apo-B ligand function can also be destroyed y
oxidation of the protein
Functions of PPAR
Nullifies inflammatory action of nuclear factor
kappa B (NFkB), a nuclear receptor normally at
rest in the cell but may be activated during
atherosclerosis

Produce cellular or macrophage apoptosis
which dampens inflammatory changes
occurring in the vessel wall
Why is DM a risk factor for MI?
Most type 2 diabetics die of heart attack.
Peroxisome-Proliferator activated receptor (PPAR) links
Type 2 diabetes mellitus with heart attack
PPAR activity
regulation of gene expression
Apoproteins of lipoproteins Enzymes of CHO & lipid metabolism
glitazone, fibrates, statins
PUFA (omega-3 ?)
Free fatty acids
Free fatty acid uptake
Insulin resistance
Complement 3
Atherosclerosis is an
inflammation-based process
LipoProteins