LIPIDS AND THE

MEDICAL PRACTICE
2012

László Fésüs
E-mail: [email protected]
INTRODUCTION

Since the metabolism, transport, structure-building and precursor

functions of lipids is so important to proper functioning of the human
body, the imbalances or deficiencies in these processes lead to
significant pathological states including some of the major clinical
problems to be encountered by physicians: obesity, arteriosclerosis
and inflammation. The pathological basis, prevention and therapy of
such diseases can not be approached without an integrated view of
lipids, the understanding of their endogenous as well as exogenous
transport and the basic principles governing the balance between
essential physiological functions of lipids and the prevention of their
accumulation in excess.
THE MOST IMPORTANT „LIPID ISSUES”
IN MEDICINE

1. Quantity and transport of lipids (particularly

cholesterol and triacylglycerol) in the body; the
special functions of the liver
2. Possible ways of lowering blood plasma
cholesterol concentration and their molecular
explanation
3. Molecular regulation of diet-induced
thermogenesis and appetite and their relationship
to the amount of triacylglycerol in the body
4. How can steroids and aspirin inhibit the
inflammatory reactions?
OVERVIEW OF THE FLOW OF PREDOMINANT LIPIDS IN
THE HUMAN ORGANISM
In meal-eating organisms such as the human, excess calories are ingested in the
anabolic phase of the feeding cycle, followed by a period of negative caloric
balance when the organism draws upon its carbohydrate and fat stores.
Lipoproteins mediate this cycle by transporting lipids from the intestine as
chylomicrons, and from the liver as very low density lipoproteins (VLDL), to most
tissues for oxidation, other functions and to adipose tissue for storage. Lipid is
mobilized from adipose tissue as free fatty acids (FFA) attached to serum albumin.
The major transport pathways are shown on the Figure presented next.

For an overall picture one should add

- details of the digestion and absorption of lipids,
- the structure and function of plasma lipoproteins,

- steps of conversion of carbohydrates to lipids in the liver and adipose tissue,

- mobilization and oxidation of fatty acids for energy production in tissues,
- production and utilization of ketone bodies from fatty acids,
- synthesis of cholesterol in the liver and in almost all cells.

These aspects are covered either in other chapters of lipid metabolism or in

sections of this chapter
 TRANSPORT OF LIPIDS IN THE ORGANISM

TAG

ApoA-I, A-II, ApoB-100, B48, ApoE, C-II: protein components of lipoproteins TAG: triacylglycerol
PLASMA LIPOPROTEINS
The central carriers of lipids around the body. Multi-component complexes of
protein and lipids forming distinct molecular aggregates and held together by
non-covalent forces.

Correspondence of plasma lipoprotein density classes with electrophoresis

mobility in a plasma electrophoresis. Albumin carries fatty acids mobilized from
adipose tissue
CHEMICAL COMPOSITION OF DIFFERENT
PLASMA LIPOPROTEIN CLASSES

(fatty acids))
LIPOPROTEINS AND THE ARTERIAL WALL

LDL and HDL particles

can reach tissue cells
SCHEMATIC REPRESENTATION OF LIPOPROTEINS

Structural studies indicate

that the inside of a lipo-
protein particle contains
neutral lipids such as
cholesterol esters and
triacylglycerols. On the out-
side of this inner neutral
lipids, in a shell about
20-Angstrom thick, reside
the proteins and the
charged amphoteric lipids.
STRUCTURAL REPRESENTATION OF A PORTION OF
THE APOLIPOPROTEIN APO C-I

Usually, the polar faces

apolipoproteins show ionisable
acid residues in the center
and basic residues at the edge.
On the other side of the helix,
the hydrophobic residues
form a nonpolar longitudinal
face.

Lys

Glu

Arg
SCHEMATIC REPRESENTATION OF A LIPOPROTEIN
 triacyllycerol Apo-C
cholesterol

esterified
cholesterol Apo-B-48

phosholipids

Its composition strongy corresponds to the lipid composition of the consumed meals
Apo-B100
 + + +
+ + ++
Positive
charges
on the surface

+ Positively charged amino acids on the surface

APOLIPOPROTEINS:
Make the existence of lipoproteins possible
Transport of lipids
Ligands for cells surface receptors
Co-factors of enzymes (e.g. C-II for lipoprotein lipase, A-I for lecithin-cholesterol-acyltransferase)

• Apo A-I HDL, chylomicron 28 KD

• Apo A-II HDL, chylomicron 17 KD
• Apo A-IV chylomicron, then HDL 46 KD
• Apo A-V 32 KD

• Apo B-100 VLDL, IDL, LDL 550 KD

• Apo B-48 chylomicron, chylomicron remnant 260 KD

• Apo C-I VLDL, HDl, chylomicron 7 KD

• Apo C-II VLDL, HDL, chylomicron 8 KD
• Apo C-III VLDL, HDL, chylomicron 8 KD

• Apo D HDL subfraction 20 KD

• Apo-E VLDL, HDL, chylomicron, chylomikron remnant 34 KD
Exist in a gene cluster in chromosome 11q23

Apo-B-100 is possibly the longest single polypeptide chain known having 4563 amino acids.
Apo-B (48% as large as B-100 is formed from the same mRNA as Apo-B 100. Apparently , in the
intestine, a stop codon - that is not present in genomic DNA is introduced by an RNA-editing
mechanism - stops translation at amino acid residue 2153 to liberate apo-B-48.
Lipoprotein a:
- Homology to plasminogen
- Protease does not activate it
- 28% is carbohydrate
- Number of kringle-4
domens is highly variable

A macromolecule complex found in human plasma that combines structural elements

from the lipoprotein and blood clotting system and that is associated with premature
coronary heart disease and stroke. Plasma Lp(a) concentrations vary 1000-fold
between individuals
COMPOSITION OF PLASMA LIPOPROTEINS IS IN
CONTINUOUS CHANGE IN THE CIRCULATION

• Enzymes
Lipoprotein lipase (LP)
anchored to the surface of endothelial cells
Lecithin cholesterol acyltransferase (LCAT)
operates in blood plasma

• Lipid transfer proteins:

Cholestrol ester transfer protein (CETP)
highly glycosilatedated 53 kDa protein
synthesized in many tissues

Phospholipid transfer protein

• Shedding and uptake of apoliproteins: from one lipopotein to the other

I. Lipoprotein lipase (on the inner surface of endothelíal cells):
Cleavage of triacylglycerol from chylomicron and VLDL

Lecithin-cholesterol acyltransferase (in blood plasma)

Transfer of fatty acid from lecithin of lipoprotein surface to cholesterol
(esterification)

II.
 Hyperlipidemias
Hyperlipidemias are disorders of the rates of synthesis or clearance of lipoproteins
from the bloodstream. Usually, they are detected by measuring plasma triglyceride
and cholesterol and are classified on the basis of which class of lipoproteins is
elevated.
• Type I accumulation of chylomicrons LPL deficiency, ApoCII
deficiency
• Type II elevated LDL LDL receptor deficiency

• Type III abnormalities of ApoE decreased uptake of IDL

• Type IV high VLDL, most common obesity, diabetes, alcohol abuse.

In diabetes insulin deficiency
causes excessive mobilization of
fatty acids and underutilization of
chylomicrons and VLDL leading to
triacylglycerolaemia

• Type V high chylomicron triacylglycerol pancreatitis, eruptive xanthomas

Hypolipoproteinemias
• Abetalipoproteinemia lack of Apo B-100 gene no chylomicron, VLDL, LDL
• Tangier disease ABCA1 deficiency HDL is 1-5% of its normal value
 TRANSPORT OF LIPIDS IN THE ORGANISM

TAG

ApoA-I, A-II, ApoB-100, B48, ApoE, C-II: protein components of lipoproteins TAG: triacylglycerol
DIGESTION AND ABSORBTION OF LIPIDS.
THE EXOGENOUS PATHWAY
An adult man ingests about 60-100 g of fat per day: more than 90% is triacylglycerol.
Additionally, 1-2 g cholesterol and 4-5 g lecithin are secreted as constituents of bile.
Lipase
acid stable form in the tongue, but mainly from pancreatic acinar cells
Reaction:
Triacylglicerol + 2H2O= β-monoacylglycerol + FFA

Bile acids inhibit lipase. Colipase (a 12 kD proteinalso secreted from the pancreas and activated by
trypsin) can neutralise bile acid action by binding to lipase ; this is critical for lipase activation (see
next figures)

Cholesteryl esterase: Cholesteryl-ester + H2O=Cholesterol +FFA

Phospholipase A: Phosphogliceride+H2O=α-acyl-lysophosphogliceride+FFA
ROLE OF PANCREASE LIPASE

2-monoacyl-glycerol

From Mol. Biol. Lecture 2008; Peter Bagossi

 STRUCTURE OF PANCREASE LIPASE

lid active center

(Ser-His-Asp catalytic triad)

N-terminal domain C-terminal domain

(a/b hydrolase fold (b-sandwich)
PDB CODE: 1N8S From Mol. Biol. Lecture 2008; Peter Bagossi
 STRUCTURE OF LIPASE-COLIPASE COMPLEX
IN THE PRESENCE OF EMULSION DROPLETS

colipase

PDB CODE: 1ETH From Mol. Biol. Lecture 2008; Peter Bagossi
 ACTIVATION OF PANCREASE LIPASE

HERMOSO ET AL. (1997) EMBO J., 16, 5531-5536 From Mol. Biol. Lecture 2008; Peter Bagossi
ACTIVATION OF PANCREASE LIPASE

- N-terminal domain contains

the calatytic site for lipolysis

- closed lid domain sterically

blocks the entry of active site

- C-terminal domain provides

the major binding site for colipase

- colipase binding leads to

attachment of lid and an open lid
domain. This and the rearrangement
of the β5 loop opens up the active
site (an oxy-anion hydrophobic hole
with the active site Ser residue)

- newly opened hydrophobic surface

areas provide binding sites for
emulsion droplets
MIXED MICELLES

Bile acids create the mixed micelles

which are crucial for concentration
of lipids and their passive uptake by
enterocytes

One of the most important

bile adcids

derived
from bile acids
UPTAKE OF LIPIDS THROUGH ENTEROCYTES.
Diffusion down a concentration gradient.
Effective concentration (maybe 1000 fold over solution of individual fatty acids) in
mixed micelles is critical; this is made possible by the packaging function of bile
salts!

Uptake of fat soluble vitamins is not possible without the micelles!

triacylglycerol triacylglycerol
cholesterol cholesterol
cholesteryl-ester cholesteryl-ester
cholesterol
cholesteryl-ester

cholesterol
CHOLESTEROL UPTAKE INTO ENTEROCYTES
Niemann-Pick C1-like 1 (NPC1) is a polytopic transmembrane protein (NPC1) that localizes
on the brush border membrane of the small intestine. It is essential for dietary cholesterol
absorption and biliary cholesterol reabsorption. NPC1 recycles between the plasma membrane
(PM) and the endocytic recycling compartment. Cholesterol depletion causes the transport of
NPC1 toward the PM. Cholesterol replenishment induces the endocytosis of NPC1L1 with
abundant cholesterol, which is dependent on the clathrin–coated complexes. In this way, NPC1
mediates cholesterol uptake through vesicular endocytosis. ACAT: esterifies cholestrol using
free fatty acids. ABCG5/G8: ABC trasporters pumping a large portion of cholesterol back into
the intestine.
EZETIMIBE
ABCG5/G8: ABC transporters
which can pump cholesterol
Back into the intestinal lumen.

ACAT2: esterifies cholesterol.

MTB: miccrosomal triacylglicerol

Transzfer protein. • Ezedoc
• Ezetrol
FA: fatty acids C:cholesterol • Zetia

Flotillins are essential for NPC1-mediated cellular cholesterol uptake and biliary cholesterol
reabsorption. Together with NPC1, they form cholesterol-enriched membrane microdomains,
which function as carriers for bulk of cholesterol. The hypocholesterolemic drug ezetimibe
disrupts the association between NPC1 and flotillins, which blocks the formation of the
cholesterol-enriched microdomains.
FATE OF CHYLOMICRONS
apo-lipoprotein
TG: triacylglycerol
C: cholesterol
P: phospholipids
CE: cholesteryl ester

CE

CE

CE

LRP1
The uptake of the remnant chylomicrons becomes possible because their
size gets small and can cross the capillary walls between endothelial cells
in the liver, then bind to chylomicron remnant receptors (LDL receptor
related protein 1, LRP1) on the surface of the hepatocytes. They contain a
small amount of triacylycerol and all the lipids which were absorbed in the
intestine, which means that, for example, all the absorbed cholesterol
reaches the liver.
The hepatocytes, depending on the dietary lipid amounts, synthesize de
novo the needed phospholipids, and cholesterol. Triacylglycerol from the
diet and what is synthesized in the liver from carbohydrate sources will be
put together, with cholesterol and phospholipids, into VLDL and secreted
into the circulation. Some part of cholesterol is secreted into the bile either
as cholesterol or bile acids made from cholesterol
 TRANSPORT OF LIPIDS IN THE ORGANISM

TAG

ApoA-I, A-II, ApoB-100, B48, ApoE, C-II: protein components of lipoproteins TAG: triacylglycerol
VLDL CATABOLISM
THE ENDOGENOUS LIPID PATHWAYS
Metabolic fate of VLDL, production of LDL.
Note that apo-E is the highest affinity ligand for LDL receptor; therefore,
the reuptake of LDL by the liver is significant

TG: triacylglycerol
C: cholesterol
P: phospholipids

CE

CE

LDL CE
(Apo-B-100)
receptor
From circulating VLDL lipoprotein lipase continuously releases fatty acids
for tissues while it becomes more and more dense, changing to first IDL,
then to LDL. It receives apolipoproteins from HDL and the plasma LCAT will
esterify its cholesterols with fatty acids cleaved from phospholipids.

LDL contains only cholesterol,

cholesterol esters and
phosphopholids carrying the
apopliprotein B (ApoB100)
Those tissue cells which
require cholesterol and
phospholipids express LDL
receptor which bind ApoB100
and take up LDL. The excess
LDL and even IDL are taken
back by LDL receptors in the
liver, with particularly the high
affinity when apoE is bound
(mainly in case of IDL)
 TRANSPORT OF LIPIDS IN THE ORGANISM

TAG

Reverse cholesterol
pathway

ApoA-I, A-II, ApoB-100, B48, ApoE, C-II: protein components of lipoproteins TAG: triacylglycerol
METABOLISM OF HDL
The reverse cholesterol transport.

A major function of HDL is to act as a repository for apolipoproteins C and E that are
required in metabolism of chylomicrons and VLDL.

The ABCA1 transporter pumps out excess cholesterol from cells

Note the involvement of LCAT in the removal of excess un-esterified cholesterol from
lipoproteins and tissues, while HDL changes from discoidal nascent HDL with
phospholipid bilayer to mature particle with the monolayer of surface lipids.

SRBI: Scavenger receptor BI.

HDL and its lipid contents, including cholesterol, will reach the liver and is taken up
by Scavenger Recetor B1 (SR-B1) of the hepatocytes.

HDL delivers high amounts of cholesterol to endocrine organs (including the adrenal
cortex) for steroid hormone synthesis.
 METABOLISM OF HDL

SR-BI

SR-B1: E
Scavenger C
Receptor BI

TG: triacylglycerol
SR-BI C: cholesterol
CE: cholesterol ester
For synthesis P: phospholipids
of steroids in
endocrine
organs
THE MOST IMPORTANT „LIPID ISSUES”
IN MEDICINE

1. Quantity and transport of lipids (particularly

cholesterol and triacylglycerol) in the body; the
special functions of the liver
2. Possible ways of lowering blood plasma
cholesterol concentration and their molecular
explanation
3. Molecular regulation of diet-induced
thermogenesis and appetite and their relationship
to the amount of triacylglycerol in the body
4. How can steroids and aspirin inhibit the
inflammatory reactions?
Cholesterol is a Janus-faced molecule. The very property that makes it essential
in cell membranes, namely its absolute insolubility in water, also makes it lethal.
When cholesterol accumulates in the wrong place, for example within the wall of
artery, it cannot be readily mobilized and its presence eventually leads to the
development of an atherosclerotic plaque. The potential for errant cholesterol
deposition is aggravated by its dangerous tendency to exchange passively
between blood lipoproteins and cell membranes. If cholesterol is to be
transported safely in blood, its concentration must be kept low.
Cholesterol is the most highly decorated small molecule in biology.
Thirteen Nobel Prizes have been awarded to scientists who devoted major
parts of their careers to cholesterol. Its complex four-ring structure and its
synthesis from a simple two-carbon substrate (acetate) through the action
of at least 30 enzymes has attracted the attention of organic chemists and
biochemists. Physiologists an cell biologists have been fascinated with its
essential role in membranes of animal cells (where it modulates fluidity, the
formation of lipid rafts and maintains the barrier between cell and its
environment) and because it is the raw material for the manufacture of
steroid hormones and bile acids. The discovery of the LDL receptor and
the relationship between its regulation and that of cholesterol synthesis
was a milestone in biological research.
These days the major focus on cholesterol by both the medical research
community and the lay public is because it is the central feature of two
major afflictions, atherosclerosis and gallstones. Curiously, although no
one can deny that a major component of most gallstones and most
atheroma is cholesterol, its pathogenetic role in these processes is
believed by many to be guilt by association rather than by causation. Only
further research efforts, particularly those applying advanced techniques of
molecular cell biology and genetics can settle the debate over this crucial
issue of current medicine.
MAJOR FEATURES OF
CHOLESTEROL HOMEOSTASIS:

• Most cells can and do synthesize it

• The average diet provides a large daily supply of cholesterol

• Cholesterol is not catabolized but excreted through the bile unchanged

or as bile acids

In addition to cholesterol several other, essential molecules are formed

at the mevalonate pathway. For this reason it is very important not to
block this synthesis pathway too much to avoid side serious effects.
SYNTHESIS

Overview of the mevalonate pathway

and its major regulatory elements.
Ensure proper balance between
endogenous synthesis and uptake
through the LDL receptor

High affinity enzymes

ensure that these
essential compounds
are always formed
in living cells

UPTAKE
REGULATION OF THE MEVALONATE PATHWAY
AT THE LEVEL OF HMG-CoA REDUCTASE
A 200-FOLD POSSIBILITY FOR REGULATION
LEVEL OF STEROL NON TARGET Magnitude of
CONTROL STEROL CHANGE

Transcription Promoter of
+ - 8X
the reductase
gene

Translation 5’ region of
- + the reductase
5X
RNA

Protein Transembrane
+ + 5X
catabolism region of the
reductase in
the
endoplasmic
reticulum
THE NUMBER OF RECEPTORS ON THE CELL SURFACE IS
THE OTHER DETERMINANT OF THE INTRACELLULAR
LEVEL OF CHOLESTEROL
Structure of the LDL receptor
In the ligand binding domain each
of the seven 40-amino-acid
repeats contains six cysteine
residues – all disulfide-bonded,
tightly cross-linked.
On each repeat there is a cluster
of negatvely charged amino acids
for binding of ligand, that is LDL,
which has positively charged
resideus on its surface .

Tyr-Val-Pro-Ala (ensures binding to clathrin and

Clustering in coated pits)
STRUCTURE OF THE LDL RECEPTOR GENE

reflecting the multi-domain structure of the protein

45 kb
18 exon
CONSEQUENCES OF CHOLESTEROL UPTAKE INTO CELLS

-- -
- ++ -
+
+ +
+
 1985

"for their discoveries concerning the

regulation of cholesterol metabolism"

Nobel Price in Physiology or Medicine

Michael S. Brown Joseph L.

Goldstein
1/2 of the prize 1/2 of the prize

USA USA

University of Texas University of Texas

Southwestern Medical Southwestern Medical
Center at Dallas Center at Dallas
Dallas, TX, USA Dallas, TX, USA

b. 1941 b. 1940
MECHANISM OF TRANSCRIPTIONAL REGULATION
BY CHOLESTEROL
There are sterol regulatory elements (SRE) in the 5’ flanking regions of genes for
HMG-C0A synthase, HMG-CoA reductase and the LDL receptor
Transcrition is regulated by proteins designated sterol-regulatory element-binding
proteins (SREBP) that are originally bound to the membranes of endoplasmic
reticulum and nuclear envelope. The known SREBPs, designated SREBP-1 and
SREBP-2, are closely related in sequence and are about 1150 amino acids in length.
The NH2-terminal region of these proteins are classic transcription factors of the
basic helix-loop-helix-leucine zipper family. The proteins are oriented so that NH2-
terminal and COOH-terminal regions both extend into the cytoplasm and a small loop
between the two membrane attachment is believed to protrude into the lumen of
endoplasmic reticulum. SREBP-1 mainly regulates genes of fatty acid synthesis after
proteolytic cleavage SP1, SP2). In sterol-depleted cells a specific cysteine protease
clips SREBP-2 at a site between the leucin zipper and the first membrane attachment
region. This free the HN2-terminal segment, which enters the nucleus and binds to
sterol regulatory elements in the promoters of the LDL receptor and HMG-CoA
syntase as well as HMG-CoA reductase genes and others. The SREBP-2 activates
transcription by virtue of acidic domains at their NH2 termini. When sterols
accumulate in cells, cleavage of both SREBPs is reduced, and any residual SREBP-
2 in the nucleus is rapidly degraded by a proteloytic enzyme different from the
cysteine protease responsible for the release of the transcriptional factor from
SREBPs. It has been presumed that a protein cofactor anchors SREB-2 to
cholesterol rich lipid rafts; after dropping of intracellular cholesterol both are released
from the endoplasmic reticulum to the Golgi where the proteolytic cleavage takes
place.
MECHANISM OF TRANSCRIPTIONAL REGULATION
BY CHOLESTEROL
Transcriptional
Cholesterol-depleted cells: activation
N
C Increased number
S2P of LDL receptors
SREBP-2 on the cell surface
N
Golgi
C N
SCAP SRE
C LDL receptor gene
ER Nucleus
S1P
Insig
The Insig protein can not keep SREBP-2 in a membrane environment where it resist to proteolytic cleavage
No transcriptional
Cholesterol-loaded cells: activation
Cholesterol Decreased number
S2P
SREBP-2 of LDL receptors
C on the cell surface
N
C
Golgi
SCAP SRE
S1P LDL receptor gene
Insig
ER C
Nucleus
C
The Insig protein brings, with the help of SCAP, SREBP-2 into a lipid raft where it can not be cleaved
STEROL REGULATORY ELEMENTS (SRE-S)
in the 5’ region of the HMG-CoA synthase, HMG-CoA reductase, LDL receptor genes.
Therefore, the intracellular cholesterol level regulates each three genes and low level of cholestereol in the
intracellular memberanes switches on HMG CoA reductase (and synthase) as well as the LDL receptor genes.

SREBP-2 fragment
DNA binding domain
Overview of the mevalonate pathway
and its major regulatory elements.

High affinity enzymes

ensure that these
essential compounds
are always formed
in living cells
 LDL RECEPTOR
GENETIC DEFECTS:
Heterozygotes
1:500
Cholesterol goes up by 2.5X
Myocardial infarction/Stroke
at the age of 35-45 years

Homozygotes
1:100 000
cholesterol goes up by 6-7X
Myocardial infarction/Stroke
at the age of 5-10 years

Type of Synthesis Transport Binding

mutation

1. no synthesis
2. not transferred to the membrane
3. binding to LDL in not efficient
4. no recycling
ARTERIOSCLEROTIC LESION ALMOST COMPLETELY
CLOSING A CORONARY ARTERY
THE CENTRAL ROLE OF LIVER IN
CHOLESTEROL HOMEOSTASIS
• Chylomicron remnant receptors are in large excess (there is no limit for
cholesterol uptake!)
• Synthesis of VLDL (with added cholesterol, about 600 mg daily)
• Largest pool of LDL receptor (re-uptake)
• Accepts HDL cholesterol (about 1300 mg daily with cholesterol re-uptake)
• Esterification and storage
• Synthesis of bile acids from cholesterol (about 250 mg daily)
• Direct secretion of cholesterol into bile (about 550 mg loss daily including
lost as bile acids)

CM-C: chylomicron cholesterol

CMr-C: chylomicron remnant
cholesterol
Dietary cholesterol can lead to
LRP1 LRP1
arteriosclerosis in two ways:
1. Direct effect of CMr
2. Reaching the it decreases
LDL receptor level

It should be noted that cholesterol secreted

into the bile and kept soluble by secreted
Phospholipids protect bile ducts from side
effects of bile acids
VLDL SYNTHESIS, IDL AND LDL RE-UPTAKE
IN THE LIVER

Apo-B
To low affinity
LDL receptors

To high affinity
Liver LDL receptors Apo-E
Apo-B

Capillary

Lipoprotein lipase
Free fatty acids
 ATHEROSCLEROSIS DEVELOPING ON GENETIC
AND DIETARY BACKGROUND
sclerotic vessel walls
C, c: cholesterol

C
C
C C CC
C
C

c
c

Because of the genetic defect of the LDL receptor The high level of intracellular cholesterol (C)
IDL and LDL are not taken up by the liver, LDL leads to down-regulation of the
level is high and from that too much cholesterol is LDL receptor level because very low
depositied into the vesel wall.
amount if SREBP2 can be cleaved
Based on genetic deficiencies in (see previous part). As a result, LDL level is
LDL receptor functions only about 5% of high (no uptake in the liver)
arteriosclerosis cases can be explained and from that too much LDL cholesterol
How can we explain the remaining 95% is deposited into the vessel wall.
Loss of function mutations in ApoE, ApoB and other molecular elements
CORRELATION BETWEEN PLASMA CHOLESTEROL
LEVEL AND THE NUMBER OF LDL RECEPTORS
IN THE LIVER
relative number of LDL receptors

100
adult Increased coronary risk
animals
80 and
newborn
60 humans

40 normal
adults
heterozygotes
20

0
homozygotes

0 1 2 3 4 5 6 7 8 9 14 16
200 mg/dl (USA)
LDL-cholesterol level (mM)
mmol/L
CORRELATION BETWEEN ESTIMATED
LDL CHOLESTEROL LEVEL OF PEOPLE IN A
COUNTRY AND THE NUMBER OF DEATH FROM
MYOCARDIAL INFARCTION (1990)

Since then prevention

measures resulted in
a very significant drop
of coronary death rates
in the USA and Finland.
Low cholesterol diet!!!!
There are many individuals who eat high fat diets but escape
hypercholesterolemia and symptomatic atherosclerosis. Within industrialized
societies there is little difference in the diets of whose suffer myocardial
infarctions and those who do not. Nearly everybody is on a high fat diet.
Clearly, atherosclerosis cannot be caused by dietary cholesterol alone.
Genetic accomplices must be implicated. The identification of mutations and
deletions in the LDL receptor gene has provided the first potential genetic
element. However, only about 5% of all cases have LDL receptor defect. The
rest of the cases also has to be explained. Recent results of molecular
genetics have identified damages of a series of other genes which lead to
hypercholesterolemia and atherosclerosis. These data have been supported
by parallel studies with knock out mice (deletion of genes):
Loss of function mutations of apo-B, apoliproteins in HDL as well as gain of
function mutations of CETP lead to the increase of the proatherogenic apo-B
containing lipoproteins.

Based on this, one may presume that a significant per cent of the
so called high fat diet hypercholesterolaemias and reéated atherosclerosis
cases are related to genetic predipositions

Significance of POLIMORPHISM
Some recent developments have opened yet another perspective in understanding
cholesterol homeostasis, namely the issue of cholesterol excretion from cells. New
members of the ATP-binding cassette (ABC) family of transporters have been
identified which transport cholesterol from cells. ABCA1 was found mutated in
Tangier disease which is characterized by defective efflux of cholesterol from cells
resulting in an inability to make high density lipoproteins.

Active transporters consist of either a single polypeptide with two ABC domains and
12 transmembrane spanning helices (full transporters, such as ABCA1) or two
polypeptides each with one ABC domain and six transmembrane spanning helices
(half transporters located within intracellular membranes).
ABCG5 and ABCG8 help to pump a large proportion of absorbed cholesterol (and
plant sterols like sitosterol) into Golgi from where ABCA1 pumps these out of
intestinal cells back into the intestinal lumen.
In inherited sitosterolemia (which is also characterized by high LDL cholesterol
concentration in the circulation because of high level of cholesterol in hepatocytes
and consequent low LDL receptor numbers) either the ABCG5 or the ABCG8 gene is
defective.

Interestingly, the nuclear receptor LXR regulates the intracellular level of these
transporters. The natural ligands for LXR are oxysterols, hydroxylated forms of
cholesterol, and in response to oxysterols/cholesterol the expression of the
transporters is increased leading to higher cholesterol efflux from peripheral tissues
as well as from hepatocytes into the bile and pumping dietary cholesterol back into
the intestine (more dietary cholesterol is re-excreted)
ABC TRANSPTORTERS INFLUENCING
INTRACELLULAR CHOLESTEROL LEVELS

The genes
of these
transporters
are regulated by
LXR which has peripheral ABCA1
Tissues
ligands derived
from ABCA1
hydroxylated
derivetives
of cholesterol
ABCG5 Passive diffusion
Sitosterol:
plant cholesterol, ABCG8
under normal Intestinal
conditions the whole lumen
amount is pumped
back into the
intestinal track.
ABCA1
POSSIBLE WAYS OF DECREASING PLASMA
CHOLESTEROL CONCENTRATION
- HDL, the anti-atherogenic lipoprotein
- Diet., significant results have been achieved with dietary restrictions!
- Blocking enterohepatic recirculation of bile acids; this leads to lower level of intracellular
cholesterol in the liver (because more bile acid have to be synthesized from cholesterol), LDL receptor gene
transcription is increased, more LDL receptors result in more uptake (lowering) of blood plasma LDL

- Inhibition of HMG-CoA reductase by STATINS. Big clinical trials show significant results!
This leads to lower level of intracellular cholesterol in the liver, LDL receptor gene transcription is increased
as a result of increased SREBP2 cleavage, more LDL receptors lead to more uptake (lowering) of blood
plasma LDL

- Gene therapy
(liver transplantation)
in homozygous LDL receptor Most of the time
deficiency STATINS are taken
without trying to block
LDLrec bile acid recircirculation.
Statin may be combined
STATINS with ezetimibe to block
cholesterol absrobtiion
through NPC1.

Cholestyramine
Colestipol
MACROPHAGE SCAVENGER RECEPTORS
collecting oxydized/acetylated LDL particles formed at high LDL cholesterol
Macrophages has very few LDL receptors but can take up oxidized/acetylated LDL particles
through their scavenger receptors and may become foam cells. Although the scavenger
receptor may play protective role in most tissues, in atherosclerotic plaque it may cause
damage. Uptake of oxidized lipoproteins may trigger the macrophage to secrete cytokines,
growth factors that stimulate collagen secretion, promote smooth muscle proliferation and
further increases LDL oxidation – all of which would increase the mass of the plaque.

Models of predicted structures of the type I (left)

and Type II (right) macrophage scavenger receptors.
C-terminal cysteine-rich domain. The collagen-like +
domain comprises 24 Gly-X-Y repeats. This model +
+ ++
assumes that the α-helical coiled-coil of domain IV +
forms a single triple-stranded left-handed superhelix
that merges with the right-handed collagen-like
triple-helix of domain V to form a single long fibrous
stalk. The positively charged C-terminal portion of the
collagenous domain is necessary for ligand binding.
All known scavenger receptor ligands are polyanions,
although not all polyanions are ligands. Scavenger
receptors work like vacuum cleaners to stuck up
oxidized lipoproteins (oxydized LDL formed
At high LDL plasma levels) and other polianions
that are stuck to collagen.
THE MOST IMPORTANT „LIPID ISSUES”
IN MEDICINE

1. Quantity and transport of lipids (particularly

cholesterol and triacylglycerol) in the body; the
special functions of the liver
2. Possible ways of lowering blood plasma
cholesterol concentration and their molecular
explanation
3. Molecular regulation of diet-induced
thermogenesis and appetite and their relationship
to the amount of triacylglycerol in the body
4. How can steroids and aspirin inhibit the
inflammatory reactions?
Obesity is a major health problem in developed countries and a growing one in
the developing world. It increases the risk of diabetes, heart disease, fatty liver
and some forms of cancer. A better understanding of the biological basis of
obesity should aid its prevention and treatment.
The terms obesity and overweight refer to excess in body
weight relative to height. The body mass index (BMI) is well
correlated with measures of body fat and is defined as weight
divided by height in meters on the second order (M2).
Overweight: 25-30 BMI. Obesity: more than 30 BMI.

The body fat, the stored mass of triacylglycerol, is in the

adipose tissue widely distributed in the body, in muscles, under
the skin, around vessels, and in the abdominal cavity, 11 kg
in normal average adult human.

The cause of most cases of obesity is not known; genetic

factors seem to have dominant role. Recent studies point to
the importance of dysfunctions in “diet induced thermogenesis”
and of several genes regulating food intake.
DIET INDUCED THERMOGENESIS, THE BROWN
ADIPOSE TISSUE AND OBESITY
Thermogenesis in brown adipose tissue. Activity of the respiratory chain produces
heat in addition to translocating protons. These protons dissipate heat when
returned to the inner mitochondrial compartment via thermogenin (uncoupling
protein, UCP-1, mitochondrial proton channel) instead of generating ATP when
returning via the F1 ATP synthase.

The passage of H+ via thermogenin, a protein dimmer of 32-kD subunit that is

absent in the mitochondria of other tissues whereas constitutes up to 15% of brown
fat inner mitochondrial membrane proteins, is inhibited by purine nucleotides when
brown adipose tissue is un-stimulated, under the influence of norepinephrine, the
inhibition is removed by the production of free fatty acids (FFA) and acyl-CoA.

Brown adipose tissue is extremely active in hibernating animals and in newborns.,

adipocytes in this tissue, unlike in white adipose tissue, contain numerous
mitochondria whose cytochromes cause its brown color. It is present in normal
individuals to varying degress, where it appears to be responsible for “diet induced
thermogenesis”, which may account for how some persons can “eat and not get fat”

Recently, two additional uncoupling proteins (UCP-2 and UCP-3) have been
identified. These are present in a number of tissues (including white adipose tissue,
skeletal muscle, kidney) where they may make contribution to energy expenditure by
heat production. In some obese persons UCPs don’t function properly because of
genetic reasons.
 DIET-INDUCED
Sympathetic
stimulation THERMOGENESIS
leptin can also
Initiate this
(see it later)

A very recent discovery:

:
UCP-1 Adult humans contain brown
adipose tissue (~ 60 gr)!
„Uncoupling protein” Behind the muscle of the lower
Thermogenin
neck and collarbone, along the
Thermogenin spine of the chest and the
abdomen.
It responds by increased heat
production to cold, sugars; it can
burn the equivalent of 4 kilograms
of triacylglicerol in a year.
Human brown adipose tissue from perithyroid region

Svensson et al., 2010, Int J Mol Med

„IRISIN, LIGHT MY FIRE”
Exercise may increase the expression FNDC5 though PGC-1α
and from FNDC5 a peptid called Irisin is released
by proteolysis. Transcriptional
Irisin can change master regulator of
white adipocytes mitochondrial genes
to brown-like
(browning).

Decrease of obesity and insulin resistance

Induction of „browning”

Kelly DP, Science 2012

OBESITY GENES
Several monogenic murine obesity models have been recently
characterized by molecular terms. The obesity associated with agouti-yellow
(Ay) mouse appears to be the consequence of ectopic expression of a
secreted protein encoded by the Ay locus. The mutations underlying the fat
phenotype has been shown to be within the gene encoding
carboxypeptidase E, a prehormon-processing enzyme.
Classical parabiosis experiments suggested that the ob encodes a
circulating factor that db may encode the receptor for the OB protein. The
cloning of these genes appears to confirm these predictions., ob encodes a
secreted protein (leptin) that function to signal fat depot levels mainly to the
brain and the db locus encodes a high affinity receptor for leptin (OB-R)
expressed abundantly in the hypothalamus. The product of the tubby obesity
gene is also expressed mainly in the hypothlamus, a region of the brain
involved in body weight regulation.
From the mouse studies the function of a so called
adipostat system has emerged. The adipostat
responds to peripheral signals, leptin that circulates
at blood levels closely related to the amount of fat stores.
Leptin is synthesized in adipose tissue regulated by the
amount fat. It reaches the central regulatory circuit of
the hypothalamus where interacts with its receptor.
The stimulation of the OB-R leads to the inhibition of
the biosynthesis and release of neuropeptide Y
(and perhaps others) in the hypothalamus. Physiologically,
neuropeptide Y (NPY) seems to have various functions. One of these is regulation of
food intake, it is the most effective neuropeptide inducing feeding behaviour and
stimulating appetite.

Another consequence of the stimulation of the leptin receptor is signalling to the sympathetic
nervous system to increase thermogenesis and energy expenditure as fat mass increases.

It has been also revealed that the action of insulin is parallel with the effect of leptin in the brain
magnifying it in the regulation of food intake. It must be noted that insulin plays a role in the
release of leptin from the fat cells, too.
THE ROLE OF NUCLEUS ARCUATUS NEURONS
IN THE REGULATION OF FOOD INTAKE
Neuroanatomical model for the demonstration of complex
regulation of food intake. Leptin and insulin stimulate the
catabolic pathways and inhibit the anabolic ones acting through
the nucleus arcuatus (ARC). These signal pathways reach the
central autonomous regulatory center( NTS) through other
neuronal (PVN, PFA, LHA). To NTS additional afferent signals
arrive from the liver and the gastrointestinal tract., these include
released peptide like CCK acting through the vagus nerve.
ARC: Nucleus Arcuatus
NTS: Nucleus Tractus Solitarius

_
+

Ghrelin

Cholecystokinin
G and CCK peptides act in short term:
empty versus full stomach
FURTHER MOLECULAR REGULATIONS OF
APPETITE AND GASTROINTESTINAL MOTILITY
The hypothalamic action of leptin also involves the melanocyte
stimulating hormone (α-MSH) and its receptor (MCR) to which the
agouti peptide (AgRP) can bind inhibiting, thereby regulating its activity.

The lack of α-MSH or MCR or the overproduction of AgRP lead to

obesity.

Leptin can decrease the production of NPY and AgRP (NPY/AgRP

neurons) and stimulate the synthesis of α-MSH (POMC neurons) in the
hypothalamus which decrease appetite.

Long-lasting or pathologic decrease of appetite leads to anorexia

Pathologic: when somebody feels/thinks Pathologic: when somebody feels/thinks
hunger while the fat cell mass is not that the fat cell mass is high and
decreased, even increased increasing while it does not, even decreases
It is very likely that the leptin system has been originally developed not for
the avoidance of obesity but rather to send signal to the body about the size
of the fat store, namely, whether it is sufficient or not for performing basic
physological functions such as growth and reproduction. For example,
ovulation stops when the mass of fat drops under a critical level and growth
slows down in children under such circumstances.

Obesity very often develops when the „adipostat” is not properly set for
various reasons. There is a pathologic starvation reaction while there is no
starvation in the body. Under such condition, sometimes even at high
leptin/insulin levels the NPY/AgRP neurons are activated and the α-MSH
producing POMC neurons are inhibited This means that the level of NPY
and AgRP is increased, AgRP further inhibits the already down-regulated α-
MSH pathway and the food intake is increased. The sustained and
pathologic increase of food intake leads to obesity. Broader metabolic
interactions are emphasized by the observation that glucocorticoids are
endogenous antagonists of leptin and insulin in the control of energy
homeostasis.

The other extreme of pathologic regulation is when the complex

neuroendocrine system signals that there is an excess of fat while this is not
the case, even the opposite may happen. In this case the result is chronic
loss of appetite, pathologic loss of weight called anorexia – which may lead
to even death.
For scientists who have proposed that obesity is the result of a
dysregulated system linking the brain, food ingestion, and
adipose tissue mass, the discovery of leptin in 1994 has
provided a very important link. The first human studies,
however, clearly showed that there is still a lot to learn.

Although in some obese patients the genetic deficiency of leptin was

observed, it turns out that serum leptin concentrations and the levels
of leptin mRNA in adipocytes in obese humans are usually elevated,
and there is a strong positive correlation between serum leptin
concentrations and the percentage of body fat and the body-mass
index. The adipocytes of humans produce excess leptin when the
adipose mass increases but there is resistance to the action of leptin,
so that the increase in adipose-tissue mass is maintained. Whether
the insensitivity to leptin is due to the insufficient transfer of leptin
through the blood-brain barrier, the mutations of the leptin receptor
genes in the brain, post-receptor abnormalities in leptin signal
transduction, or other abnormalities in hypothalamic function is
unknown.
The control and regulatory processes of food intake and energy expenditure
are very complex and even after the significant new discoveries their details
and precise mechanism are still uncovered.

Very recently a new peptide, CART (cocaine- and amphetamine- regulated

transcript) has been found in the brain which inhibits the action of NPY, the
synthesis of CART is induced by leptin.

Several other neuropeptides have been found in the hypothalamus which

either increase (galanin, orexin A, B) or decrease (CRH, TRH) appetite.

Cannabinoids increase appetite and the synthesis of their endogenous forms

is inhibited by leptin.

There are several parallel and interacting regutarory pathways which

determine food intake and energy expenditure at the level of the central
nervous system.
MOLECULAR REGULATION OF APPETITE

Increases appetite
NPY
AgRP

Ghrelin, motilin

galanin
orexin
Cannabinoids (endogenous, too)

Decreases appetite
Leptin: decreases NPY level in the hypothalamus decreases AgRP level
(as a result α-MSH can work) increases CART level
(which decreases NPY level)

α-MSH

Cholecystokinin
PYY3-36 (from colon); long term effect
Based on the above detailed new biomechanical informations
pharmaceutical companies have started to develop drugs
influencing steps of the regulatory pathways of food intake.
Seeing the complexity of these processes it is very likely that the
combination of the compounds will lead to results in treatment of
obesity and reaching optimal weight control

The importance of such efforts is emphasized by the very high

risk of the occurance of diabetes, cardiovascular diseases and
hypertonia in obese people.
Recent results point to the importance of resistin, a peptide
released from increased mass of fat and causing insulin
resistance, in development of type II diabetes.
THE MOST IMPORTANT „LIPID ISSUES”
IN MEDICINE

1. Quantity and transport of lipids (particularly

cholesterol and triacylglycerol) in the body; the
special functions of the liver
2. Possible ways of lowering blood plasma
cholesterol concentration and their molecular
explanation
3. Molecular regulation of diet-induced
thermogenesis and appetite and their relationship
to the amount of triacylglycerol in the body
4. How can steroids and aspirin inhibit the
inflammatory reactions?
Linoleic and linolenic acids are not synthesized in the human body
and must be provided in the diet. The essential fatty acids give rise to
polyunsaturated C20 (eicosanoic) fatty acids from which are
synthesized very important groups of physiologically and
pharmacologically active compounds, the eicosanoids. The dietary
supply of essential fatty acids is delivered to cells by the plasma
lipoproteins from the liver where part of it is converted to eicosanoic
fatty acids, such as arachidonic acid (enzymatic steps of conversion
are shown in an earlier chapter).

Cells build the essential fatty acids, including arachidonic acids, into
their membrane phospholipids where they play a significant role in
maintaining fluidity.

Arachidonate and other eicosanoic fatty acids are usually in the 2-

position of phospholipids and released after various stimuli of cells as
a result of phospholipase A2 activity, they become substrates for the
synthesis of several compounds. The two major types of oxidized
derivatives, collectively called eicosanoids, are prostanoids and
leukotrienes which may belong to three groups. In addition, epoxy-
eicosatrienoic (EETs) and autooxidation products may be also
formed (not shown).
THE THREE GROUPS OF EICOSANOIDS
AND THEIR BIOSYNTHETIC ORIGIN

NORMAL DIET

„ESKIMO” DIET

The subscript denotes the total number of double bonds in the molecule.
THE CYCLOOXIGENASE
AND LIPOXYGENASE
PATHAYS

Numbers represent enzymes of which

phospholipases (1), cyclooxigenase (COX)
1 or 2 (2) and lipoxygenase (9,12,13) and
various types of prostaglandin (4-8) and
leukotriene (10, 11) synthases are
especially important
Eicosanoids are formed by virtually every tissue
in the body. The biological activities of eicosanoids
vary from organ to organ and may differ markedly
even in different phases of organ function.
They are short living substances (PGs minutes,
LTs hours) acting at very low concentrations
and usually in a paracrine fashion. Various stimuli of
a cell (e.g. angiotensin, bradykinin, epinephrine,
thrombin) may lead to release of membrane
arachidonate and the production PGs which leave
the cell and act on receptors of another cell.
PHYSIOLOGICAL EFFECTS OF PROSTAGLANDINS
CYCLOOXIGENASE 1 (COX-1) EFFECTS

• Inhibition of HCl secretion in the stomach (PGE2)

• Contraction of the uterus (PGF2α)
• Initiation of sleep (PGD2)
• Keeping open the ductus arteriosius in the fetus (PGE2)
• Vascular system:
Thromboxanes formed in patelets (aggregation of
platelets,contraction of smooth muscle)
PGI2 formed in the endothelial cells (inhibition of aggregation of
platelets, inhibition of contraction of smooth muscle)
• Cellular differentiation, migration, others

A very recent discovery: in white adipose tissue PGE2 and PGI2 levels are
increased during adrenergic stimulation and they increase the expresssion
of brown adipose tissue genes which lead to production of heat from
fatty acid oxidation.
The prostaglandin receptors are very specific. They share
structural features of G protein associated seven
transmembrane receptors. Specificity is achieved by their
spectra of isoforms, differences in tissue distribution and using
different signal transduction pathways.
- Five basic types of prostanoid receptors (DP, IP, FP, TP) and
four subtypes of PGE receptors (EPI-4) have been cloned and
characterized.
- Functionally significant topographical patterns of receptor
distribution have ben delineated in single organs such as the
brain.
- Signalling: in smooth muscle cells each type of receptor that
mediates contraction transduces an increase in Ca2+ and each
that induces relaxation signals increase in cAMP.
VASCULAR EFFECTS OF PROSTAGLANDINS

Thromboxanes are synthesized in platelets and upon release

vasoconstriction and platelet aggregation. Prostacyclins (PGI2)
are produced by endothelial cells and are potent inhibitors of
platelet aggregation. Thus, the two are antagonists.

The low incidence of heart disease, diminished platelet

aggregation, and prolonged clotting times in Greenland
Eskimos have been attributed to their high intake of fish oil
containing eicosapentaenoic acid (EPA) which give rise to the
series 3 prostaglandins (PG3) and thromboxanes (TX3). PG3
and TX3 inhibit the release of arachidonate from phospholipids
and the formation of PG2 and TX2. PGI3 is as potent an anti-
aggregator of platelets as PPGI2, but TX2 is a weaker
aggregator than PGX2, thus the balance of activity is shifted
toward non-aggregation
INFLAMMATION, PAIN AND FEVER ELICITED BY PGE2
MEDIATED BY CYCLOOXIGESA 2 (COX-2) INDUCED IN
CELLS DURING INFLAMMATION

Inflammtory
cytokines

In the hypothalamus
induced COX-2 heat regulatory center
PHARMACOLOGICAL SIGNIFICANCE OF
THE CYCLOOXYGENASE PATHWAY
The effect of steroids (e.g. glucocorticoids)
The intracellular protein, lipocortin ( the synthesis of which is stimulated by
glucocorticoids and other steroids at the transcription level acting through their
nuclear receptor) can prevent release of eicosanoic fatty acids from membranes
by inhibiting phospholipase A2: this explains the anti inflammatory effect of
steroids.

Mechanism of action of non-steroidal anti-inflammatory compounds

The non-steroidal anti-inflammatory compounds, represented here by aspirin,
inhibit cyclooxygenases. It should be noted that there are two isoforms of
cyclooxigenase: COX-1 (expressed constitutively) and COX-2 (induced during
inflammation). Aspirin inhibits COX-1 more efficiently than COX-2 and the side
effects of aspirin treatment (like lethal bleeding in the stomach) are mainly
explained by COX- inhibition. Future drug developments will aim specific
inhibition of COX-2 to achieve effective inhibition without major side effects.
 Steroids
Steroids
through changing
expression of genes
(increased lipocortin expressiion)

COX-1/2

Phospho -
lipase A2
LEUKOTRINES ARE POTENT REGULATORS OF
MANY DISEASE PROCESSES
The slow reacting susbstance of anaphylaxis (SRS-A) is a mixture of
leukotriene C4, D4, and E4. This mixture of leukotrienes is formed in
released from mast cells upon stimulation and is 100-1000 times more
potent than histamine or prostaglandins as a constrictor of the bronchial way
musculature. Both these and leukotriene B4 also causes vascular
permeability and attraction as well as activation of leukocytes.

Receptors for LTC4, LTD4, LTB4 and LXA4 have been defined
pharmacologically and biochemically and shown to be G protein- associated
membrane proteins.

The formation of leukotrienes can be blocked by steroidsot but not by

aspirin
Phospho -
lipase A2
Inhibition
by inducing
lipocortin glutathion

Steroids
THE MOST IMPORTANT „LIPID ISSUES”
IN MEDICINE

1. Quantity and transport of lipids (particularly

cholesterol and triacylglycerol) in the body; the
special functions of the liver
2. Possible ways of lowering blood plasma
cholesterol concentration and their molecular
explanation
3. Molecular regulation of diet-induced
thermogenesis and appetite and their relationship
to the amount of triacylglycerol in the body
4. How can steroids and aspirin inhibit the
inflammatory reactions?
 The most important questions
1 Basic characteristics and function of the major lipoproteins (chylomicron, VLDL, LDL
and HDL).
2. What are the functions of apolipoproteins?
3. What leads to changes in composition of lipoproteins in the circulation?
4. How lipids (fatty acids and cholesterol) enter into the intestinal epothelial cells and then get
into the blood plasma?
5. The exogen lipid transport - major features
6. The endogen lipid transport - major features
7. The reverse cholesterol transport - major features
8. Regulation of the mevalonate metabolic pathway, what are the products??
9. How does LDL cholestrol get into cells?
10. How increased intracellular cholesterol concentration regulates the LDL receptor level:
significance of SREBP2.
11. What genetic defects may lead to high (pathologic) LDL cholesterol level?
12. How cholesterol rich diets lead to pathologic level of LDL cholesterol?
13. What is the relationship between the mass and function of brown adipose tissue and body
weight?
14. Explain the regulatory consequences of increased and decreased leptin concentration in the
circulation?
15. In addition to leptin what other molecules you know which increase or decrease appetite?
16. How prostaglandins are generated and what are their physiologic effects?
17. What is COX-2, when does it function and what is consequence of its activation?
18. Molecular explanation of the effects of aspirin and steroids?