FEDERAL UNIVERSITY LOKOJA

CHOLESTEROL METABOLISM/
LECTURE SERIES 5 IN MBCH 212 (Metabolism of
Lipids)

Assoc. Prof. Bamidele Ajilore (MBChB; PhD)

CONTACTS: +2347033985727
[email protected]
Department of Medical Biochemistry
Federal University Lokoja
Nigeria 1
 Course Synopsis
1. Fate of dietary lipids; Transport of lipoproteins; Role of lipoprotein lipase; Clinical importance of
lipoprotein transport.

2. Metabolism of Triglycerides in adipose tissue; Metabolism of fatty acids in the liver: ketogenesis
& complete oxidation to carbon dioxide and water.

3. Metabolism of fatty acids between adipose tissue and liver; Transport vehicles: role of very low-
density lipoprotein (VLDL) particles; Lipid transport (clinical importance of lipid transport);
Hormonal influence on fat metabolism

4. Fatty acid metabolism in feed and fasting state; Complete oxidation of fatty acids

5. Metabolism of cholesterol: Synthesis of cholesterol from acetyl-CoA. Site of synthesis

(cytoplasm & Endoplasmic reticulum); Role of HMG CoA reductase; Cholesterol transport: Role
2
of LCAT (Lecithin Cholesterol Acyl Transferase).
LEARNING OUTCOMES- At the end of the lecture, students should be able to:

o revise their knowledge of chemistry of cholesterol.

o learn how cholesterol is synthesised in the body? Note the starting material required for
cholesterol biosynthesis.
o list the tissues in which cholesterol biosynthesis occurs.
o learn in detail the various steps in cholesterol biosynthesis and the enzymes and
required.
o know what is HMG-CoA. How it is formed? Learn the fates of HMG-CoA—why it is called
“committed step”?
o describe the “rate-limiting” step in biosynthetic pathway and study how the cholesterol
biosynthesis is regulated
o explain the metabolic fate of cholesterol in the body.
o explain the role of cholesterol and TG in atherosclerosis and IHD

3
 BRIEF OVERVIEW OF STEROLS AND
CHOLESTEROL CHEMISTRY
Steroids are complex molecules containing 4 fused rings

Steroids are often found in association with fat but may be

separated from the fat, after the fat is saponified, since they
occur in unsaponifiable residue

All of the steroids have a similar cyclic nucleus resembling

phenanthrene (ring A, B and C) to which a cyclopentane ring
(ring D) is attached.

It is designated as cyclopentano perhydro-phenanthrene

nucleus

Other names for cyclopentano perhydro-phenanthrene ring

are:
Sterane ring or Gonane ring
4
When the hydrogen atom at position 3 on the ring is replaced by hydroxy group, we
have a steroid alcohol called sterols.

Methyl side chains occur typically at positions 10 and 13 (constituting carbon atoms
19 and 18 respectively) and a side chain at position 17.

Sterols are therefore alcohols of gonane

They are amphipathic lipids synthesized from acetyl coenzyme A via HMG-CoA
reductase pathway

Sterols occur naturally in eukaryotes including plants, animals and fungi.

 Sterols of plants are called phytosterols and sterols of animals are called zoosterols.

The most important zoosterol is cholesterols.

5
 Notable phytosterols include: campesterol, sitosterol and stigmasterol

 Sterols play important role in the physiology of eukaryotic organisms

 For example, cholesterol is an important part of animal cell membrane where it

affects the cell membrane fluidity and serve as secondary messenger in
developmental signaling.

 In humans, corticosteroids e.g. cortisol acts as signalling compound

 Sterols are common components of human skin oils

 Phytosterols have been shown in clinical trials to block cholesterol absorption in

human intestine, thereby helping to reduce cholesterol absorption in humans
6
 At present, American Heart Association has recommended that supplemental plant
sterols be taken by patients with hypercholesterolemia (but to be avoided in pregnant
women and nursing mothers)

 Some preliminary researches have shown that phytosterols may have anti-cancer effects
Cholesterol
Cholesterol is the most important sterol in human body. Its molecular formula is
C27H45OH.

It is biosynthesized by all animal cells and is an essential structural component of animal
cell membrane

7
 It possesses “cyclopentanoperhydrophenanthrene nucleus”.

 It has an –OH group at C3.

 It has an unsaturated double bond between C5 and C6.

 It has two –CH3 groups at C10 and C13.

 It has an eight carbon side chain attached to C17.

 The name cholesterol is derived from the Greek word meaning solid bile.

 It occurs as a white or faintly yellow, almost odourless, granules.

8
 It is insoluble in water, sparingly soluble in alcohol and soluble in ether, chloroform,
hot alcohol, ethyl acetate and vegetable oils.

 It easily crystallises from such solutions in colourless, rhombic plates

 It is not saponifiable. Its melting point is 147 to 150o C.

 Since it has an unsaturated bond, it can take up two halogen atoms.

 Source:
i. Exogenous: Dietary cholesterol, approximately 0.3 gm/day. Diet rich in cholesterol
are butter, cream, milk, egg yolk, meat, etc. A hen’s egg weighing 2 oz gives 250 mg
cholesterol

ii. Endogenous: Synthesised in the body from acetyl CoA, approximately 1.0 gm/day
9
 Cholesterol is found in largest amounts in normal human adult:

 brain and nervous tissue 2 per cent,

 in the liver about 0.3 per cent,
 skin 0.3 per cent and
 intestinal mucosa 0.2 per cent,
 certain endocrine glands, viz. adrenal cortex contain some 10 per cent or more, corpus
luteum is also rich in cholesterol.

 The relatively high content of cholesterol in skin may be related to vit D formation by
UV rays

 and that in the adrenal gland and gonads to steroid hormone synthesis.

 Cholesterol is present in blood and bile and is usually a major constituent of gallstones
10
Site of Synthesis-
Enzymes- Enzyme system
 Essentially all tissues form
involved in cholesterol
cholesterol
biosynthesis are associated
 Liver is the major site of
with:
cholesterol biosynthesis • Cytoplasmic particles
 other tissues are active in BIOSYNTHESIS OF “microsomes”
this regard, e.g. adrenal CHOLESTEROL • Soluble fraction—cytosol
cortex, gonads, skin, and
intestine are most active
 Low order of synthesis: Starting Material-
 Principal precursor is ‘Active’
Adipose tissue, muscle,
aorta and neural tissues. acetate (acetyl-CoA)
 Brain of new-born can Acetyl CoA formed in - The entire carbon skeleton, all
glycolysis and Oxidation of 27 C of cholesterol in humans
synthesize cholesterol fatty acid are the
while the adult brain precursors for the can be synthesised from active
cannot synthesize cholesterol synthesis acetate.
cholesterol. 11
 V. Conversion of Lanosterol Cholesterol

IV. Cyclisation of Squalene Lanosterol

III. Formation of Squalene (30C) Condensation of 6

isoprenoid units
II. Formation of “Iso-Prenoid units”(5C) + PO4 to Mevalonate
with loss of CO2
I. Synthesis of mevalonate (6C)
2 Acetyl-CoA (3C)

Steps of Biosynthesis of Cholesterol

12
I. Synthesis of Mevalonate from
Acetyl-CoA Consists of two steps:

 Formation of HMG-CoA: (β-OH-β-

methyl glutaryl-CoA): This step also
occurs in 2 stages
• Most important and rate limiting
step and irreversible reaction

• Enzyme contains—SH group

NADPH required as cofactor
supplied by HMP Pathway.

• Hormones: Insulin and thyroid • Dietary cholesterol and

hormones— increases reductase endogenously synthesised
 In the next step, which is the rate- activity; Glucagon and cholesterol inhibits this ‘rate-
limiting step, HMG-CoA is converted glucocorticoids—reduces the limiting’ step (“feedback”
to Mevalonic acid (Mevalonate) activity. inhibition).
catalysed by the enzyme HMG-CoA
• Also inhibited by cyclic AMP
reductase.
13
 III. Formation of squalene:
II. Formation of Isoprenoid Units:  The pyrophosphorylated isoprenoid units condense
to form ultimately a 30-carbon aliphatic chain called
 Mevalonate is phosphorylated by ATP to Squalene. The condensation occurs in three steps:
form several ‘active’ phosphorylated
intermediates. o One molecule of iso-pentenyl-pyrophosphate first
condenses with one molecule of 3,3-dimethyl allyl
 Three such phosphorylated compounds are pyrophosphate to form a 10-C compound called
formed and it is followed by decarboxylation geranyl pyrophosphate, the reaction is catalysed by
to form first “active” iso-prenoid unit: ”Iso- the enzyme geranyl pyrophosphate synthase.
pentenyl pyrophosphate” (5 C). One of
intermediate phosphorylated compound is  Another molecule of iso-pentenyl pyrophosphate
“Mevalonate-3-phospho-5-Pyrophosphate” reacts with geranyl pyrophosphate to form the 15C
which is unstable. compound farnesyl pyrophosphate, the reaction is
catalysed by the enzyme “farnesyl pyrophosphate
 Iso-pentenyl pyrophosphate undergoes synthetase”.
isomerisation to form another 5 C iso-
prenoid unit, called “3-3’-Dimethyl allyl  Finally, 2 molecules of farnesyl pyrophosphate
pyrophosphate”. condenses to form 30C aliphatic called squalene,
reaction is catalysed by the enzyme squalene
synthetase. 14
IV. Cyclisation of squalene to form lanosterol: V. Conversion of lanosterol to cholesterol:
Main changes that are brought about are:
 The formation of lanosterol from squalene takes
place in two steps:  Removal of three angular –CH3 groups. This
involves a series of reactions, mechanism of
• In the first step squalene-2,3-epoxide is formed demethylation is not properly known. CH3
catalysed by the enzyme squalene mono-oxygenase; group at C14 is first eliminated.
which requires NADPH and molecular O2.
 Shift of double bond between C8 and C9 to C5
• In the next step, an enzyme cyclase brings about the and C6, and
cyclisation of squalene to form lanosterol.
 Saturation of double bond in side chain.

15
Synthesis of Isoprenoid Units

16
 Synthesis of Squalene Formation of Lanosterol

2 possible pathways have been suggested for the conversion of lanosterol to cholesterol:
17
 Control of Cholesterol Biosynthesis

I. SREBP Pathway:
• This is a feedback control- The end product
cholesterol controls its own synthesis of the enzyme
by a feedback mechanism.

• Increase in the cellular concentration of cholesterol

reduces the synthesis of the enzyme by decreasing
the transcription of the gene responsible for the
production of HMG CoA reductase.

• This occurs at the irreversible stage of cholesterol

biosynthesis and it called the rate limiting step or
committed step.

• Repression of transcription of the “HMG-CoA

reductase” by cholesterol is via activation of a “sterol
regulatory element-binding protein” (SREBP)
transcription factor. 18
II. Hormonal regulation: Role of cyclic AMP:
• The HMG CoA reductase exists in two • HMG-CoA reductase may exist in active/inactive forms,
interconvertible forms. which is reversibly modified by phosphorylation/and
dephosphorylation mechanisms. This is mediated by c-
• Insulin and thyroid hormones Increase HMG AMP dependent protein kinases.
CoA reductase activity.
• Cyclic AMP inhibits cholesterol biosynthesis by
• The dephosphorylated form of the enzyme is converting “HMG-CoA reductase” to inactive form.
more active, phosphorylated is less active.
Effects of starvation/fasting:
• Glucagon and glucocorticoids decrease HMG- • Fasting and/starvation also inhibit the enzyme and
CoA reductase activity. activate “HMG-CoA lyase” to form ketone bodies.
• Hormones exert their influence through cAMP
Effect of feeding cholesterol:
• Inhibition by drugs: The drugs Compactin and • Feeding cholesterol reduces the hepatic biosynthesis
lovastatin, mevastatin, simvastin are of cholesterol by reducing the activity of “HMG-CoA
competitive inhibitors of HMG CoA reductase. reductase”.
It is used to decrease the cholesterol
• Intestinal cholesterol biosynthesis does not respond
• HMG CoA reductase is inhibited by bile acids. to the feeding of high cholesterol diets.
19
20
 Transport of Cholesterol
• Cholesterol in the diet is absorbed from the • Highest proportion of circulating cholesterol is
intestine, and in company with other lipids, found in LDL (β-lipoproteins) which carry
are incorporated into “chylomicrons” and also cholesterol to tissues and also in HDL, which
to some extent ‘VLDL’. takes cholesterol to liver from tissues for
degradation
• Of the cholesterol absorbed, 80 to 90 % in the
• Dietary cholesterol takes several days to
lymph is esterified with long-chain FA.
Esterification may occur in the intestinal equilibrate with cholesterol in plasma, and
mucosa. several weeks to equilibrate with cholesterol in
tissues.
• In man, the total plasma cholesterol varies
• Free cholesterol in plasma and liver equilibrates
from 150 to 250 mg% (average 200 mg%),
rising with age, although there are wide in matter of hours.
variations between individuals.
• In general, free cholesterol exchanges readily
• The greater part is found in the ‘esterified’ between tissues and lipoproteins, whereas
form and is transported as “lipoproteins” in cholesterol esters do not exchange freely
plasma.
21
• Some plasma cholesterol ester may be formed in HDL as a result of trans-esterification reaction in plasma
between cholesterol and FA in position-2 of lecithin which is catalysed by the enzyme lecithin-cholesterol
acyl transferase (LCAT).

Factors that decrease Cholesterol in Cells:

Factors that increase Cholesterol in Cells: i. Efflux of cholesterol from cells to HDL
(scavenging action).
i. Increased synthesis of cholesterol.
ii. Esterification of cholesterol by the
ii. Hydrolysis of cholesterol ester by the enzyme enzyme AcylCoA-cholesterol acyl
“cholesterol ester hydrolyase”. transferase (ACAT).

iii. Uptake and delivery of cholesterol in cells by iii. Utilisation of cholesterol for synthesis of
circulating LDL (uptake by specific receptors). steroid hormones, viz. glucocorticoids,
mineralo-corticoids, Gonadal hormones.
iv. Uptake of cholesterol containing lipoproteins
by ‘non-receptor’ mediated pathway. iv. In liver cells: formation of cholic acid.

v. Uptake of free cholesterol by cell membranes v. Formation of vit D3

22
Role of LCAT:

• High density lipoprotein (HDL) and the enzyme lecithin-cholesterol acyl

transferase (LCAT) are responsible for the transport and elimination of
cholesterol from the body.

• LCAT is a plasma enzyme, synthesized by the liver.

• LCAT catalyses the transfer of fatty acid from the second position of
phosphatidyl choline (lecithin) to the OH group of cholesterol.

• HDL cholesterol is the real substrate for LCAT and this reaction is freely
reversible.

23
 OTHER FACTORS THAT INFLUENCE CHOLESTEROL LEVEL IN BLOOD

2. Dietary cholesterol:
1. Dietary fats: • Increased feeding of cholesterol in diet decreases
• Increased intake of fats in the diet increases endogenous synthesis and reduces cholesterol level.
level of cholesterol by increased synthesis.
• It is difficult to lower the normal blood cholesterol
• Greater amount of saturated fatty acids level by taking food of low cholesterol.
increases cholesterol level
• Restricted dietary intake of cholesterol is usually
• Substitution in the diet of saturated FA by balanced by increased biosynthesis.
polyunsaturated FA has beneficial effect and
lowers cholesterol level: 3. Dietary carbohydrates:
i. It stimulates oxidation of cholesterol to bile
acids • Increased consumption of carbohydrates increases
ii. Stimulation of cholesterol excretion in cholesterol level.
intestine
iii. Shift cholesterol from plasma to tissues • Consumption of excessive amount of sucrose and
iv. Cholesterol esters of polyunsaturated FA are fructose cause increase in plasma lipids particularly
more rapidly metabolised by liver and other TG and also cholesterol.
tissues
24
4. Heredity: 8. Minerals:
• Hereditary factors play the greatest role in • In vitro acetate to cholesterol conversion in tissue
determining individual blood cholesterol cell cultures depressed↓ by addition of Vanadium
concentrations. and increased by chromium and Manganese salts.

5. Blood groups: • Conversion of mevalonate to cholesterol was

• Cholesterol level found to be slightly higher inhibited by Vanadyl SO4.
in persons belonging to blood groups ‘A’ and
‘AB’, than those belonging to ‘O’ and ‘B’ 9. Dietary fibres:
groups. • Increased fibres in the diet, caused an increased
excretion of cholesterol and bile acids in faeces in
6. Calorie Intake: experimental animals and produced significant
• Intake of excess calories increases reduction in serum cholesterol.
cholesterol level.
10. Physical exercise:
7. Vitamin B-Complex: • Studies on human volunteers showed hard physical
• Nicotinic acid: In large doses has cholesterol exercise brought about lowering in serum
lowering effect. cholesterol↓ level and increased level of HDL ↑.
• Pyridoxine deficiency: Produces increase in
blood cholesterol level and atherosclerosis
in monkeys. 25
11. Life style of the individual:
- Life style of the individual also affects Factors leading to higher or fluctuating levels of FFA
serum cholesterol level. Additional factors include:
which play a part in coronary heart disease
include: • Emotional stress
• Obesity • Nicotine from cigarette smoking
• Lack of exercise and sedentary habits • Coffee drinking and
• Smoking • Partaking a few large meals rather than more
• High blood pressure continuous feeding.
• Associated diabetes mellitus
• Drinking of soft as opposed to hard water. Pre-menopausal women appear to be protected against
these deleterious factors, probably due to the hormones,
12. Elevation of Plasma FFA: estrogens and high HDL, as compared to men and
• Elevation of plasma FFA due to any cause postmenopausal women.
will enhance increased VLDL secretion by
the liver by enhancing endogenous TG Hypolipidaemic drugs:
synthesis leading to increased cholesterol • Several drugs are known to block the formation of
output to the circulation cholesterol at various stages in the biosynthetic
pathway, or increase the catabolism/excretion of
cholesterol. Many of the drugs also have harmful side
effects. 26
 Hypolipidemic Drugs Mechanism of Action
1. Nicotinic acid: In large doses has Reduces the flux of FFA by inhibiting adipose tissue lipolysis,
hypocholesterolaemia effect thereby inhibiting VLDL production in liver. In large doses, may
produce fatty liver
2. Oestrogen
Lower cholesterol level and increases HDL
3. Sitosterol
Acts by blocking esterification of cholesterol in gut thus reducing
cholesterol absorption (Synthesis may be increased later on)
4. Dextrothyroxine (cholaxin),
and Neomycin Increases faecal excretion of cholesterol and bile acids

5. Clofibrate and Gemfibrozil a. Inhibits secretion of VLDL by liver b. Inhibiting hepatic cholesterol
synthesis c. Probably also increases faecal excretion d. They facilitate
6. Resins, e.g. Colestipol, hydrolysis of VLDL triacyl glycerol by lipoprotein lipase
Cholestyramine (Questran)
Prevent the reabsorption of bile salts by combining with them,
increasing their faecal loss
7. Statins e.g. Atorvastatin,
Simvastatin, Pravastatin All statins inhibit HMG-CoA reductase-enzyme, thus up-regulating LDL
receptors reduces LDL-cholesterol level 27
 METABOLIC FATE OF CHOLESTEROL RELATION OF CHOLESTEROL AND OTHER LIPIDS AS
RISK FACTOR IN CORONARY HEART DISEASE (CHD)
 Cholesterol is converted into the compounds
 Of the serum lipids, cholesterol has been the one
showed in the diagram on the next slide.
most often incriminated as the risk factor.
 Cholesterol is mainly excreted in the form of
 However, other parameters such as serum TG,
bile salts in stool.
VLDL and LDL have been incriminated.
 Increased plasma cholesterol results in the
 Patients with CHD can have any one of following
accumulation of cholesterol under the tunica
intima of the arteries causing abnormalities:
atherosclerosis.
1. Elevated concentrations of VLDL with normal
 The progression of the disease process leads concentrations of LDL
to narrowing of the blood vessels.
2. Elevated LDL with normal VLDL
 Dietary intake of polyunsaturated fatty acid
(PUFA) helps in transport and metabolism of 3. Elevation of both VLDL and LDL
cholesterol and prevents atherosclerosis
28
Fate of cholesterol in the body 29
1. Role of Cholesterol:
 An elevation of the total cholesterol in plasma is Protective mechanism is explained by:
a. Reverse transport of cholesterol from
considered to be a ‘Prime risk factor’ for CHD.
peripheral tissues into the Liver by way of
 The Framingham Study has demonstrated a linear HDL which thus reduces the intracellular
cholesterol content (scavenging action of
increase in coronary “risk” with increment of total
HDL).
plasma cholesterol level from 180 mg% upwards.

 The Lipid Research Clinics Coronary Primary b. High HDL concentrations are associated with
a faster elimination from the plasma of TG
Prevention Trial had proved that in humans, a
rich lipoproteins and their atherogenic
lowering of plasma cholesterol level reduces the
intermediates
coronary thrombosis and myocardial infarction and
mortality.
3. Role of TG and VLDL
 Elevated VLDL and hypertriglyceridaemia may
2. Role of LDL and HDL
 A raised HDL concentration is beneficial and also be considered a primary ‘risk’ factor
 A low blood TG level is suggestive of efficient
protective against CHD
intravascular lipolysis and thus of enhanced
 LDL is the carrier of 70 per cent of total cholesterol formation of HDL
 Hypertriglyceridaemia indicates less effective
and it transports cholesterol to tissues and thus is
intravascular lipolysis and hence a reduced
most potential atherogenic agent 30
formation of HDL