LIVER

BIOCHEMISTRY
 LIVER STRUCTURE

sinusoids
central vein

portal vein

bile canaliculi
bile duct
hepatic artery
 Liver functions
1.Liver is a main organ which is responsible for
dividing of nutritional substances in our organism
(for example, glucose, triacylglicerides and ketone
bodies).

2.All types of metabolism (protein, lipid,

carbohydrate, vitamins, mineral) take part in liver.

3. Hepatocytes synthesizes as lot of blood

lipoproteins, low-weight bioactive substances
(creatin, 25-oxicalciferol, heme), cholesterol.

4. Synthesis of urea (final product of nitrogen

metabolism) also takes place in the liver.
 Liver functions
5. Liver synthesizes bile acids and excrete a
bile into intestinal tract. This process play
a very important role in lipids digestion and
excretion of cholesterol and some products
of metabolism into intestine.
6. Liver play desintoxification role,
inactivates endogenic and exogenic
substances (ammonia, metabolic waste,
drugs, alcohol and chemicals, some
hormones, different toxins).
7. Liver is a depo for iron and some another
metals.
8. Liver can synthesize blood plasma
proteins and non-essential amino acids.

9. Liver synthesizes purine and

pyrimidine nucleotides, creatine,
nicotinic acid, choline, carnitine,
polyamines.
 10. Liver stores glucose in the
form of glycogen which is
converted back to glucose again
when needed for energy.

11. Liver stores the vitamins

A, D, E, B12,folate and
synthesizes blood clotting
factors.
 METABOLISM OF CARBS IN LIVER
- glycolisis
Glucose from intestine pass into the
liver, where of it undergone the
phosphorillation. Glucose-6-phosphate
formed in result of this reaction, which
catalyzed by two enzymes –
hexokinase and glucokinase.
 Metabolism of fructose and galactose
Fructose and galactose also transformed into
glucose-6-phosphate in the liver.
Glucose-6-phosphate is a key product of
carbohydrates metabolism. In the liver this
substance can metabolized into different ways
depend of liver’s and whole organism’s necessity.
1. Synthesis of glycogen.
2. Glucose-6-phosphatase catalize dephosphori-
llation of glucose-6-phosphate and free glucose
formed.
3. Excess of glucose-6-phosphate, decomposites in
glycolysis for pyruvate and for acetyl-CoA,
which are used for fatty acids synthesis.
4. Glucose-6-phosphate decomposites for H2O and
CO2, and free energy for hepatocytes formed.
5. Part of glucose-6-phosphate oxidized in
pentosophosphate cycle. This way of glucose
decomposition supplyes reducted NADPH, which
is necessary in fatty acid synthesis, cholesterin
synthesis, and also ribose-phosphates for
nucleic acids.
 gluconeogenesis
conversion of pyruvate into
acetyl CoA
 tricarboxylic acid cycle
 pentose phosphate pathway
 glycogenolysis,
glycogenogenesis
 METABOLISM OF LIPIDS IN LIVER
Lipogenesis
(synthesis of fatty acids
and lipids).
Substrate for this
process – acetyl-CoA,
formed from glucose
and amino acids.
This process is very
active when the person
eats a lot of
carbohydrates.
Liver play a central role in synthesis of
cholesterol, because 80 % of this
amount is synthesized there.
Liver is a place of ketone
bodies synthesis. These
substances from liver
transported to another
tissues, first of all to the
heart, muscles, kidneys
and brain. Кetone bodies
are main source of
energy for many tissues
of our organism during
starvation (brain).
 synthesis of lipoproteins
The major carriers of triacylgliсerols and
cholesterol are chylomicrons, very low density
lipoproteins (VLDL), high density lipoproteins
(HDL) and low density lipoproteins (LDL).
- synthesis of triacylglyserols;
- synthesis of phospholipids;
- lipolysis;
- fatty acids oxidation.
 METABOLISM OF PROTEINS IN LIVER
- In liver present all
nessesary enzymes, that
catalyse reactions of
metabolism of proteins and
aminoacids.
- In liver synthesized
100 % of albumins, 90 % of
α1-globulines, 75 % of α2-
globulines, 50 % of β-globulins,
blood clotting factors,
fibrinogen, such enzyme as
cholinesterase.
 In liver toxic ammonia in
result of protein
decomposition convert into
intoxic urea.
 - Conversion of proteins into carbs and lipids;
- Conversion of proteins into low molecular weight
nitrogen containing substances – purine and pirimidine
nucleotides, creatine, nicotinic acid, heme, choline,
carnitine, poliamins.
The most critical aspects of protein metabolism that occur
in the liver are:
Deamination - is the removal of the amino group as ammonia
(NH3) and transamination - is the transfer of an amino group
from an amino acid to keto acid. Several enzymes used in
these pathways (for example, alanine and aspartate
aminotransferases) in diagnostic of liver disease.
 VITAMIN METABOLISM IN
LIVER
• Formation of active form of vitamin D
• Formation of vitamin A from carotins
• Depo of cyanocobalamine and folic
acid
• Depo of vitamin E
• Phosphorilation of vitamins B,
formation of coenzyme forms
 The decomposition of hemoglobin in tissues
After a life span of about 120 days the erythrocytes
die. The dead erythrocytes are taken up by the
phagocytes of the reticuloendothelial system of the
body (spleen, bone marrow, liver).
The Hb molecule is broken down into 3 parts:
1. The protein (globin) part of Hb is utilized
partly in to aminoacids or along with other body
proteins.
2. The iron is stored in the reticuloendo-
thelial cells and is used for the synthesis of Hb
and other iron containing substances of the
body.
 3. The porphyrin part is converted to bile pigment, i.e.
bilirubin which is excreted in bile.
Bile pigments formation
Heme in the presence
of the enzyme, heme
oxygenase, loses one
molecule of CO and
one atom of iron in
Fe3+ form producing
biliverdin. In this
reaction, the porphyrin
ring is cleaved by
oxidation of the alpha
methenyl bridge
between pyrrole rings.
The enzyme needs
NADPH+H+ and O2.
Biliverdin which is green in color is
the first bile pigment to be produced;
it is reduced to the yellow-colored
bilirubin, the main bile pigment, by
the enzyme biliverdin reductase
requiring NADPH+H+.
Bilirubin is non-polar, lipid soluble
but water insoluble. Bilirubin is a very
toxic compound.
 Bilirubin formed in reticuloendothelial
cells then is associated with plasma
albumin to protect cells from the toxic
effects. As this bilirubin is in complex
with plasma proteins, therefore it
cannot pass into the glomerular filtrate
in the kidney; thus it does not appear in
urine. This bilirubin is called
indirect bilirubin or free bilirubin or
unconjugated bilirubin.
• Complex – bilirubin-albumin (or indirect
bilirubin) in liver degraded into bilirubin and
albumin. Albumin with blood go back to the
blood stream.
• In liver bilirubin interact with UDP-glucuronic
acid and is converted to the water soluble form -
bilirubin mono- and diglucoronids. This reaction
is catalized by UDP-glucoroniltransferase.
Another name of bilirubin mono- and
diglucoronids is conjugated bilirubin or direct
bilirubin or bound bilirubin.
Conjugated bilirubin is water soluble and is
excreted by hepatocytes to the bile.
Conjugated bilirubin or (bilirubin-diglucoronids)
undergoes degradation in the intestine through
the action of intestinal microorganisms.
From bilirubin-diglucoronids spleet glucoro-
nic acid that with blood tranported to the liver.
In small intestine bilirubin is reduced and
urobilinogen is formed. The reduction of
urobilinogen results in the formation of
stercobilinogen (in a colon). Stercobilinogen
is oxidized and the chief pigment (brown color)
of feces stercobilin is formed
 A small part of urobilinogen is
reabsorbed by the mucous of
intestine and via the vessels of vena
porta system enter to liver. In
hepatocytes urobilinogen is used for
formation of pyrol compounds which
are excreted with bile. This
urobilinogen in urine is called
urobilin, or true urobilin.
Another bile pigment that can be
reabsorbed in the intestine is
stercobilinogen. Stercobilinogen is
partially reabsorbed in the lower part
of colon into the haemorroidal veins.
From the blood stercobilinogen pass
via the kidneys into the urine where it
is oxidized to stercobilin. Another
name of urine stercobilin is false
urobilin.
• Urobilinogen and
stercobilinogen
don’t have color.
Only urobilin and
stercobilin have
yellow-orang color.
• Every day in adult
exreated 250 mg of
bile pigments with
feces and 1-2 mg
with urine.
The total bilirubin content in the
blood serum is 1,7-20,5
mcromol/l,
Indirect (unconjugated) bilirubin
content is 1,7-17,1 mcromol/l
Direct (conjugated) bilirubin
content is 0,86-4,3 mcromol/l.
 Some other differences between these two
forms of bilirubin are given below:
Property Unconjugated Conjugated
1. Solubility Soluble in water,
Soluble in lipid,
insoluble in lipid
insoluble in water
2. Excretion in Yes
No
urine

3. Deposition in
Yes No
gram

4. Plasma level
Pre-hepatic Hepatic and
is increased in
jaundice jaundice posthepatic
Jaundice or icterus is the orange-yellow
discoloration of body tissues which is
best seen in the skin and conjunctivae; it
is caused by the presence of excess of
35 mmol/l bilirubin in the blood plasma
and tissue fluids. Depending upon the
cause of an increased plasma bilirubin
level, jaundice can be classified as
1) pre-hepatic,
2) hepatic and
3) post-hepatic
JAUNDICES
 NORMAL METABOLISM OF BILE
PIGMENTS
albumin
CELLS OF RES
Indirect Indirect
bilirubin bilirubin 1,7- Indirect bilirubin
20,5 mkmol/l
UDP-glucoronil-
NADP+ Biliverdin albumin transferase
NADPH2 reductase

Bilirubin mono-
Biliverdin glucoronid, 20 %
Direct
L
Iron bilirubin 0.8- I
Globin
4.3 mkmol/l V
Bilirubin di-
glucoronid, 80 %
E
Verdoglobin
R
B Dipyrols
L
NADP+
O
Hemoxi- -glucoro-
genase O nidase
D
Glucoronic
NADPH2 acid
Hemoglobin ERYTHROCYTES
B
I
Direct L
bilirubin E

K
I I
Mesobilirubin
D N
N T
E E
Y Mesobilirubin S
(urobilinogen) T
S
I
N
E
Stercobilinogen Stercobilinogen

URINE STOOL

Stercobilin Stercobilin
 HEMOLYTIC (PREHEPATIC) JAUNDICE

Jaundice due to the excessive breakdown

of red blood cells.

Causes:
•sickle cell anemia,
•malaria,
•thalassemia,
•autoimmune
disorders,
•massive hemorrhage
METABOLISM OF BILE PIGMENTS IN HEMOLYTIC
JAUNDICE
CELLS OF RES albumin
Indirect Indirect Indirect
bilirubin bilirubin bilirubin
UDP-glucoronil-
NADP+ Biliverdin albumin transferase
reductase
NADPH2
L
Bilirubin mono- I
Biliverdin glucoronid, 20 %
Direct
V
Iron bilirubin E
Globin Bilirubin diglucoronid, R
80 %
Verdoglobin

NADP+ B
Hemoxi- L -glucoro-
genase O nidase
O
D
Glucoronic
NADPH2 acid
Hemoglobin
ERYTHROCYTES
B
I
Direct L
bilirubin E

K Mesobilirubin I
I N
D T
E
N Mesobilinogen S
E (urobilinogen) T
Y I
S N
Stercobilinogen E
Stercobilinogen

Urobilin Stercobilin
STOOL
URINE Stercobilin
Urine dark Stool hypercholic
PARENCHYMAL (HEPATIC) JAUNDICE
occurs due to the liver disease and
inability of liver to metabolize and
remove bilirubin from the biliary
system
Causes:
•cirrhosis,
•cancer,
•viral hepatitis,
•Gilbert’s syndrome,
toxins or drugs, etc.
 Hepatic jaundice is characterized by
• 1. Increased levels of conjugated and
unconjugated bilirubin in blood serum.
• 2. Dark coloured of urine due to the
excessive excretion of bilirubin and
urobilinogen.
• 3. Pale, clay coloured stools due to the
absence of stercobilinogen.
• 4. Increased activities of alanine and
aspartate transaminases.
 ОBSTRUCTIVE (POST-HEPATIC)
JAUNDICE
is caused by obstruction of bile flow from
the liver
Causes:
•carcinoma in the bile
duct or gall bladder;
•presence of
gallstones in the
biliary system;
•infection by
parasites;
•pancreatitis, etc.
 Obstructive (post hepatic ) jaundice is
characterized by
•1. Increased concentration mainly of
conjugated bilirubin in blood serum.
•2. Dark beer coloured urine due to
elevated excretion of conjugated bilirubin
and clay coloured feces due to absence of
stercobilinogen.
 Jaundice in Newborns (Hyperbilirubinemia)
Physiological jaundice typically occurs
because a newborn’s liver is immature and
processes bilirubin more slowly, because the
enzyme UDP-glucuronyl transferase is
deficient. It usually appears within a few
days after birth and resolves within two
weeks.
In rare cases, if the bilirubin level stays high and
isn't treated, it can cause brain
damage called kernicterus. This can lead to serious
lifelong problems.
DETOXIFICATION OF TOXIC SUBSTANCES IN LIVER
A xenobiotics is a compound that is foreign to the body. The
principal classes of xenobiotics:
Exogenic xenobiotics - medical relevance are drugs,
chemical cancerogens, (insecticides, herbicides, pesticides,
food additions, cosmetics, domestic chemical
substances).

Endogenic xenobiotics - some internal substances also have

toxic properties (for example, bilirubin, free ammonia,
bioactive amines, products of amino acids decay in the
intestine). Moreover, all hormones and mediatores must
be inactivated.
 Reactions of detoxification take
place in the liver. Big molecules
like bilirubin excreted with the
bile to intestine and leaded out
with feces. Small molecules go to
the blood and excreted via kidney
with urine.
Phase I:
Biological transformation of xenobiotics

 hydrolysis,
 reduction,
 oxidation.

These reactions introduce functional group (—

OH, —NH2, —SH, or —COOH) and usually
result in a little increase of hydrophylic
properties
General ways of xenobiotics biotransformation and their localization in cell

REACTION ENZYME LOCALIZATION

PHASE I
Hydrolysis Esterase Microsomes, cytosol, lysosomes, blood
Peptidase lysosomes
Epoxide hydrolase Microsomes, cytosol

Azo- and nitro-reduction Microflora, microsomes, cytosol

Reduction Carbonyl reduction Cytosol, blood, microsomes
Disulfide reduction Cytosol
Sulfoxide reduction Cytosol

Alcohol dehydrogenase
Oxidation Cytosol
Aldehyde dehydrogenase
Mitochondria, cytosol
Aldehyde oxidase
Cytosol
Xanthine oxidase
Cytosol
Monoamine oxidase
Mitochondria
Diamine oxidase
Cytosol
Flavin-monooxygenases
Microsomes
Cytochrome P450
Microsomes
PHASE II
Glucuronide conjugation Microsomes
Sulfate conjugation Cytosol, microsomes
Glutathione conjugation Cytosol
Amino acid conjugation Mitochondria, cytosol
Acetylation Mitochondria, microsomes
Methylation Cytosol, microsomes, blood
 PHASE I
Hydrolysis
Esterases (carboxyesterases, cholinesterases,
phosphatases)
Peptidases
Reduction
Metals and xenobiotics containing aldehyde, keto,
disulfide, alkyn, azo, or nitro group are often reduced
Reducing agents:
Reduced glutathione,
FADH2,
FMN,
NADH
NADPH.
 Oxidation
Alcohol dehydrogenase
 Aldehyde dehydrogenase

Oxidizes aldehydes to carbonic acids

Xanthine dehydrogenase-Xanthine oxidase

Monoaminooxidase
Oxidative deamination of amines (serotonin) and
many xenobiotics
 Cytochrom P450
The highest concentration – in endoplasmic reticulum of
hepatocytes (microsomes).
Microsomal oxidation system include cytochrom P450
and flavin enzyme P450 reductase

Hem containing protein.

Catalyzes monooxigenation of
oxygen atom into substrate;
another oxygen atom is reduced
to water
Electrons are transferred from
NADPH to cytochrome P450
through flavoprotein NADPH-
cytochrome P450 reductase.
SCHEME OF MONOOXYGENASE
SYSTEM
Phase II – conjugation
includes:
 glucuronation,
 sulfation,
 acetylation,
 methylation,
 conjugation with glutathione,
conjugation with aminoacids (glycin, taurin,
glutamic acid)

Phase II results in the marked increase of

hydrophylic properties of xenobiotic.
 Glucuronidation, the combining of
glucuronic acid with toxins, requires the
enzyme UDP-glucuronyl transferase
(UDPGT). Many of the commonly
prescribed drugs are detoxified through
this pathway. It also helps to detoxify
aspirin, menthol, vanillin (synthetic
vanilla), food additives such as
benzoates, and some hormones.
Sulfation is the conjugation of toxins with
sulfur-containing compounds. The
sulfation system is important for
detoxifying several drugs, food additives,
and, especially, toxins from intestinal
bacteria and the environment.
Sulfation is also used to detoxify some
normal body chemicals and is the main
pathway for the elimination of steroid and
thyroid hormones.
 Glutathione is also an important
antioxidant. This combination of
detoxification and free radical
protection, results in glutathione
being one of the most important
anticarcinogens and antioxidants in
our cells, which means that a
deficiency is cause of serious liver
dysfunction and damage.
 A few xenobiotics (amines, phenol, tio-
substances, inorganic compounds of sulphur,
selen, mercury, arsenic) are subject to
methylation by methyltransferases, employing
S-adenosylmethionine as methyl donor. Also
catecholamines and nicotinic acid amid (active
form of vitamin PP) are inactivated due to
methylation.
Very important way of detoxification is
ureogenes (urea synthesis). Free ammonia,
which formed due to metabolism of amino acids,
amides and amines, removed from organism in
shape of urea.
SCHEME OF MONOOXYGENASE SYSTEM IN
ENDOPLASMIC RETICULUM
The example of reaction that is catalyzed by cytochrome P450: hydroxylation of
aliphatic carbon
The example of reaction that is catalyzed by cytochrome P450: hydroxylation of
aromatic carbon
Examples of reactions catalyzed by cytochrome P450: heteroatom oxygenation
Examples of reactions catalyzed by cytochrome P450: oxidative group transfer
Bilirubin di-glucoronid