MY

UNIVERSITY
STEP INTO TOMORROW

Ms. Tayyaba Zulfiqar

Physiology-B
BLOOD
 INTRODUCTION

Definition: Blood is a specialized connective tissue in

which there is a liquid intercellular substance known as
plasma and formed elements.
It circulates in a closed system of blood vessels and
chambers of heart by pumping of heart.

Formed elements:
Red blood cells-Erythrocytes
White blood cells-Leucocytes
Platelets- Thrombocytes
Composition of blood

Plasma-
55%

Buffy coat-<1%

Formed elements-45%
 Composition of blood
SOLIDS:
A- Inorganic constituents
B- Organic constituents
C- Colored substances
D- Other substances

A-INORGANIC CONSTITUENTS
Na+ 138--- 142 mEq/L

K+ 4.5---5.5 mEq/L

Ca++ 2.4 mEq/L

Mg++ 1.2---1.5 mEq/L

Cl- 103 mEq/L

HCO-3 24-32 mEq/L ,28mEq/L

PO4 1-4 mEq/L

SO4 1 mEq/L
 B-ORGANIC CONSTITUENTS
• Plasma proteins 6.5---7.5 gm/dl
Albumin 4.5gm/dl
Globulin 2.5gm/dl
Fibrinogen 100---300mg/dl
Prothrombin 10---20mg/dl
• Non-protein nitrogenous substances
28---48mg/dl
Urea 15-40mg/dl, Uric acid 4---8mg/dl,
Creatinine 0.2---1.2mg/dl,
Amino acids 40mg/dl
Xanthine, hypoxanthine
C-COLORED SUBSTANCES
Bilirubin 0.3---1mg/dl
Carotene

D-OTHER SUBSTANCES
Hormones
Enzymes
Vitamins
Minerals
Metabolites (Lactic acid, Ketone bodies)
 Functions of Blood
1- Transport function:
A- Respiratory function:
Hemoglobin the respiratory pigment present in
the Red Blood Cells (RBCs) increases oxygen
carrying capacity of blood.
B- Nutritive function:
Glucose, amino acids, fatty acids, vitamins,
minerals & water.
• C- Excretory function: Urea, uric acid, creatinine
through kidney.
• Bile pigments through liver.
• CO2 through lungs.

• D- Distribution function: Hormones & hormone

binding proteins towards target tissues & organs.

• E- Temperature regulation: Heat produced in active

tissues in the body core is brought to the skin and
lungs. It is regulated by the amount of blood flow to
these areas.
• The water in plasma has

i) High specific heat-can absorb large amount of heat.

ii) High thermal conductivity-distribute heat.
iii) High heat of evaporation-heat loss.

2- Regulatory function:
a) Maintenance of ionic balance and internal environments
between cells & ICF.
b) Maintenance of water balance between interstitial intracellular
and vascular compartments.
c) Maintenance of acid base equilibrium. Reduced and oxidized
hemoglobin, plasma proteins with amino and carboxyl groups are
buffers.
 d) Regulation of blood volume - Hemostasis
Excessive blood loss is prevented by clotting of
blood (clot formation) by platelets and clotting
factors .
3- Defensive function:
 Neutrophils and monocytes: Phagocytosis of
dead tissues invading micro organisms.
 T lymphocytes: Cellular immunity
 B lymphocytes & plasma cells: Humeral
immunity (antibodies).
 Eosinophils: Phagocytosis of allergic complex
4- Functions due to plasma proteins
1-Proteins exerts colloidal osmotic pressure and
contribute to dynamic distribution of water between
blood, lymph and intracellular fluids.

2- Transport function

3- Nutritive function

4- Buffer function

5- Precursor of active substances

6- Hormones - angiotensinogen & angiotensin 1

7- Blood clotting factors

8- Immunity by antibodies

9- Inflammatory proteins

10- Blood pressure - due to viscosity of blood.

11- Enzymes - Amylase, lipase

 What’s in plasma?
Water (90-92%)
Solids (8-10% )
1. Nutrients
2. Plasma proteins
• Albumins
• Alpha and beta globulin
• Gamma globulin (antibodies)
-All except gamma globulins are formed in the liver,
maintain osmotic pressure (and thus blood volume)
Gases CO2 3ml/dl, PCO2 46mmHg
O2 0.3ml/dl, PO2 35mmHg
N2 0.9ml/dl
Wastes
 Difference between Plasma and
interstitial fluid

• Plasma and interstitial fluid-- their ionic

composition is similar.
• The most important difference between these
two compartments is the higher concentration of
protein in the plasma; because the capillaries
have a low permeability to the plasma proteins,
only small amounts of proteins are leaked into
the interstitial spaces in most tissues.
• The composition of extracellular fluid is
carefully regulated by various mechanisms,
but especially by the kidneys.

• This allows the cells to remain continually

bathed in a fluid that contains the proper
concentration of electrolytes and nutrients for
optimal cell function.
 Difference between serum and plasma

• Plasma and serum are both found in blood. Both are

constituents of blood

• The main difference between plasma and serum is the

absence of fibrinogen, an important clotting factor, in
the serum.

• Neither plasma or serum contain red or white blood

cells or platelets, but they contain all the other
substances, except for the fibrinogen, which is not
 Difference between human and animal
blood
• Human blood is always warm but not the blood in
all the animals except mammals and birds.
• The percentages of the cell types in humans and
other animals are different
• Humans have a closed and complete blood vessel
system, whereas some animals have open and/or
incomplete blood systems.
• The efficiency of function of the human blood is
very high, which can be compared to other animals.
 Plasma proteins-Total protein level
6.8-7.5 gm/dl
1-Albumin
2-Globulin
3-Fibrinogen—Cf---1
4-Prothrombin-Cf---11
5-Clotting factors
6-Proteins of inflammation-complement
system
7-Fibrinolytic system
8-Hormones , enzymes, antibodies
 Plasma proteins
1-Albumin:
60% 0f total PP. 4.5—5.7gm/dl
Mol.wt 69000
Water soluble
Heat coaguable
Precipitated by full saturation of (NH4)2SO4
Produced by liver cells
Responsible mainly for colliodal osmotic pressure of blood so
controls distribution of fluid into diff. fluid compartments of
body.
2-Globulin:
30% 0f total PP
1.5—2.5gm/dl
80 000---200 000---1000 000
Types α1, α2 , β , γ
Insoluble in water but soluble in dilute salt solution
It is ppted with half saturation of (NH4)2SO4
• Mainly transport function and immunity
• Mostly 50-70%produced by liver,
• γ globulin not produced by liver but by lymphatic tissue
• According to electrophoresis pattern types of globulin are:
• α1----carries thyroxine
• α2----- carries copper (Ceruloplasmin),erythropoietin,
prothrombin
• β--- carry lipid--lipoproteins, steroids, cholesterol.

γ --- produced by B lymphocytes - plasma cells and confer

humoural Immunity.

So called immunoglobulins include Ig G (75%),A,M,E,D.

(Also include blood group antibodies).
 3-Fibrinogen-Clotting factor1

• 5% of total plasma proteins.

• Amount in blood----300mg/dl
• Mol. Wt.--- 330, 000
• Produced by the liver
Is changed to Fibrin during blood coagulation
process. Forms clot , maintains haemostasis
 4-Prothrombin-clotting factor—11
• Conc. In blood=0.1-0.2gm/L
• Mol.wt 68,700
• Produced by the liver and vit K is required for its formation.
In liver diseases (for example viral hepatitis A,B,C,D and
E) prothrombin conc. falls because liver cells are damaged
and there is deficiency of vit. K.(Prothrombin time is
prolonged).
• Prothrombin is an inactive and unstable protein. During
coagulation changes into its active form the thrombin.
Blood clotting
 Functions of plasma proteins

1-Proteins exert colliodal osmotic pressure and

contribute to dynamic distribution of water bet.
blood, lymph and interstitial fluids.
• With protein conc. 7.5 gm/dl albumin exerts 70% of
total osmotic pressure.
• Regulate filteration- absorption of fluid across the
capillary membrane.
2-Proteins perform transport functions:
Albumin----carries bilirubin
α1 Globulin---- carries thyroxin, polysaccharides
(glyco proteins)
• α2Globulin----- carries copper (Ceruloplasmin),
• β ---lipoproteins carry lipid, steroids,
cholestrol.
β---globulin-- Apotransferrin--- carries iron (Fe)---
transferrin.
3-Proteins perform haemostatic functions:
Prothrombin, fibrinogen, blood clotting factors
v,vii,viii,ix,x,xi,xii and xiii are protiens which take
part in clotting of blood. Prevent haemorrhage and
bleeding when injures.
Anticoagulant or fibrinolytic system liquifies fibrin
clot.
4-Proteins perform nutritive functions:required for
repair of tissues, musculoskeletal growth and energy
purposes if carbohydrates and fats are not available.
are in a state of dynamic equilibrium with cellular proteins.
Undergo constant breakdown to supply raw material to
body tissues & constantly being replenished by synthesis
of new plasma proteins.
(Protein poor diet in infants-Kwashirkor-- (Protein –calorie
malnutrition)--important clinical features)
Kwashiorkor-protein calorie
malnutrition

A Child with Kwashiorkor.

5-Proteins perform immune functions:
Immunoglobulins (antibodies) provide resistance
against infections. (antigen antibody reactions).
• Chemically glycoproteins of mol wts 150, 000 to
1000,000.(γ globulins).

• 6-Proteins perform buffer functions:Act as an

important intracellular buffer.
• At body pH 7.4 act as anions (negatively charged)and
accept H+.1/6th of total buffering capacity of blood is
due to proteins. COOH- splits into COO- and accepts
H+ .
7-Plasma Proteins precursors of active substances:
Hydrolyse to release biologically active products.
Kininogens---- Kinins
Angiotensinogen--- Angiotensin 1
Erythropoietinogen---Erythropoietin

8-Plasma Proteins responsible for viscosity,specific gravity

of plasma and blood pressure,blood volume: sp. Gravity
of plasma=1.027, (blood=1.055---1.066.)
Viscosity of whole blood is due to RBCs and viscosity of
plasma is due to plasma proteins especially the globulins.
Relative viscosity of blood is 4.7 for men, 4.4 for females,
4.2 for children. Multiple myeloma increase abnormal
proteins, increased plasma viscosity.
• 9-Plasma Proteins include proteins of
inflammation: Increase during inflammation
protect against antigen.
• Gamma globulins
• Complement system
• Acute phase proteins
• C reactive proteins
• Fibrinogen
• Haptoglobins
• α1antiglobins
 Impact of plasma proteins on ESR

• Plasma Proteins affect ESR: Erythrocyte

sedimentation rate (ESR) is a diagnostic and
prognostic blood test.
• Albumin decreases ESR by increasing buoyancy for
RBCS and Globulin.
• Fibrinogen increase the ESR by increasing rouleaux
formation . Fibrinogen has multiple positive charges
on their surface. Neutralizes the surface negative
charge on RBCs.
 Methods of separation of plasma proteins

1) Precipitation by salts
• Salting out process. Albumin is precipitated by full
saturation of (NH4)2SO4solution. Globulin is
precipitated by half saturation. Fibrinogen by 5%
(NH4)2SO4solution.
2)Electrophoresis
• It is the movement of charged particles in an
electrical field towards the oppositely charged
electrode. Various proteins move at diff. speed and
this can be photographed.
Paper and Gel electrphoresis
3) Immunoelectropheresis: In this method
electropheretic and immunolological properties of
proteins are used to split various protein fractions.
• Proteins in agar gel first exposed to electropheresis
which separates proteins in zones along the path of
their migration.
• Anode positively charged electrode and anions
negatively charged ions move towards it.
• Then specific immune serum containing antibodies
against different protein fractions is placed in a
groove in agar gel .
Immunoelectropheresis
Immunoelectropheresis
• The sites where blood proteins and immune
antibodies come together by diffusion become
marked by precipitation lines in the gel. These lines
are photographically recorded. Useful to find
different fractions of Ig (Immunoglobulin).
4)Ultracentrifigation:
• It involves centrifugation of plasma at a very high
force of gravitational pull. Diff. proteins sediments at
different rates according to their mol. wts & densities
and thus can be separated from this mixture.
5) Cohns’ fractional precipitation method:
It is based on fractionation with low salt concentration at low
temperatures, varying pH, solvent is alcohol. Proteins are
separated due to varying solubility.

6)Kjeldahl’s method: 16 gms of Nitrogen is present in 100

grams of plasma proteins.
So 1 gm of Nitrogen is present in 6.25 gms of plasma
proteins.
Total N of blood is measured first.
Then measure Non-protein nitrogen (NPN)& subtract it from
the total nitrogen. It gives Protein N (of plasma) & conc of
plasma protein is measured by the formula:
Total protein nitrogen-Conc. of NPN=protein Nx6.25
 Biosynthesis of plasma proteins

• All plasma proteins albumin, fibrinogen,50-70% globulin

and prothrombin are synthesized by the liver cells.
• Immunoglobulins are synthesized by lymphoid tissue.
• 7-14 days are required for complete regeneration of pp.
Fibrinogen appears first then globulin, prothrombin and
albumin.
• Rate of protein formation can be very fast 30g/day.
• In infancy pp conc. is less 5-5.5gm/dl. Adult 6.9-7.8gm/dl.
• In liver disease cirrohsis, burns, kidney dis. pro. conc
decreases.
Red blood cells (Erythrocytes)
 RBC is the predominant formed element

Occupy 40-45% of blood volume

• RBC COUNT :
• In normal man 5,200,000
• In normal woman 4,700,000

• SIZE & SHAPE OF RBCS:

• Circular
• non-nucleated
• biconcave discs
• Mean diameter 7.8 µm.
• Thickest at sides 2.5 µm,
• thinner in center 1um or less.
 Shape
Bi-concave shape increase the surface area
greatly for the exchange of gases. Total surface
area of all RBCs in human is 38000 sq. meters

• The shape can be deformed temporarily as

it squeezes through capillaries 6 µm
diameter.
This process is called diapedesis.
Unique shape(biconcave) of RBC contributes in
2 ways to perform their main function of O 2
transport.

• Biconcave shape provides greater surface area

for O2 diffusion across membrane.

• The thin inner center enables O 2 to diffuse

rapidly between exterior and innermost
regions of the cell.
 Red blood cell membrane

• Red blood cell is a “bag” that can be deformed into

almost any shape
• It has a great excess of cell membrane for the
quantity of material inside, deformation does not
stretch the membrane greatly and does not rupture
the cell

• Membrane is pliable, flexible and highly elastic.

• It contains 10 major and 100 minor proteins.

 Structure of RBC membrane
Lipid bilayer(40%)
Membrane proteins (52%)
Carbohydrate (8%)
- Support structural integrity of RBC.
• Spectrin, ankyrin , protein 3 are mainly responsible for
flexibility of membrane.
• This flexibility helps RBCs to pass through capillaries.
• Sialic acid in glycocalyx give negative charge to RBC
membrane
 Hemoglobin
RBC contains hemoglobin which enables it to
transport O2. Hemoglobin is only found in
RBCs.
Hemoglobin has two parts:
1. Globin – a protein made up of highly folded
polypeptide chains.
2. Heme groups – 4 iron containing non protein
groups each one is bound to one of the
polypeptides.
 Hemoglobin
• A pigment
• Due to its iron content, it appears red when
combined with oxygen
• Bluish when deoxygenated
• Combines with Oxygen --- CO2, H ions, CO,
NO
Morphological changes of RBCs
Mature RBCs
Does not contain
• Nucleus
• Mitochondria
• Endoplasmic reticulum
• Golgi apparatus
• Ribosomes.
 Enzymes in RBCs
Mature RBCs contain two types of enzymes:
1) Glycolytic enzymes
• Essential for generating energy.
• Maintain ionic concentration of cell.
• Carry oxygen but cannot use it.
• Rely on glycolysis for ATP formation.
2) Carbonic anhydrase
• Catalyzes conversion of CO2 in HCO3 ions
which is main transport form of CO2 in blood.
 Metabolic processes of RBCs

Glucose is the only source , transported into the cell interior by

glucose transporter 1 (GLUT 1 by facilitated diffusion).
• 90% of glucose enters into anaerobic glycolysis.
• 10% of glucose is supplied to Hexose Monophosphate
Shunt (HMP) pathway or pentose phosphate pathway.
• It produces NADP+H+which keeps Glutathion reductase in
reduced form.
• This enzyme repairs the effects of spontaneous oxidation of
sulph-hydral groups of cell membrane and oxidation of Fe+
+ of heme to Fe+++.
• Deficiency of reduced Glutathion reductase leads to
oxidative damage and loss of membrane pliability—
hemolysis.
• Certain drugs like primaquin decreases reduced
Glutathion and causes hemolysis & hemoglobinuria.
 Energy is utilized in RBCs
• To drive cation pump which expels Na + out
and carries K+ inside the cell & prevents
hemolysis.
• Keep glutathione in reduced form.
• To keep heme Fe in reduced Fe++ form.
 Life span of RBC

• Total life span 120 days

• Lacks organelles for production of enzymes (proteins). Old

RBCs lose pliability become spiculated, fragmentation and
osmotic lysis occurs in spleen.

• During its lifetime it travels about 700 miles as it circulates

through vessels.
 Hemolysis
• RBC burst and release their haemoglobin.
• This Hb is phagocytized by macrophages of
body, spleen, bone marrow & kupffer cells of
liver.
• After some period, macrophages release iron
from Hb & pass it back into blood & carried
by transfferin protein.
• Iron pass to bone marrow for new production
of RBC or to liver for storage of iron that later
used for bile production for fat digestion.
 STRUCTURE OF Hb
• Hb is a conjugated protein.
• It consists of a protein combined with iron
containing pigment
 Formation of Hemoglobin
• Synthesis of hemoglobin begins in the
proerythroblasts and continues into the
reticulocyte stage
• Therefore, when reticulocytes leave the bone
marrow and pass into the blood stream, they
continue to form minute quantities of Hb.
• In mitochondria
 Formation of Hemoglobin
• 2 succinyl-CoA (formed in the Krebs
metabolic cycle) + 2 glycine

one pyrrole
• Four pyrroles 1,2,3,4 combine by CH4 bridge

protoporphyrin
IX
• Protoporphyrin IX + iron (Fe++)ferous
form

Heme molecule
• Each Heme molecule + globin
(polypeptide) synthesized by ribosomes

hemoglobin chain
• Four of these in turn bind together

loosely to form the

whole HEMOGLOBIN molecule.
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• Substances needed for synthesis of Hb:

 Proteins

 Metals----Fe,Cu,Co,Ni

 Vitamins---C, riboflavin, nicotinic acid, B6

June 30, 2024 72
• There are several slight variations in the
different subunit hemoglobin chains,
depending on the amino acid composition of
the polypeptide portion.
• The types of globin chains
 Alpha chains
 Beta chains
 Gamma chains
 Delta chains
 TYPES OF Hb
• Hb-A Adult hemoglobin
The globin contains 2 alpha & 2 beta
chains

• Hb-F Fetal hemoglobin

The globin contains 2 alpha and 2 gamma
chains
 Abnormal Hemoglobin

• Cause: Genetic mutation

• There are two types of abnormal hemoglobin:
1. Hemoglobinopathies
2. Hemoglobin in thalassemia and related disorders

 Hemoglobinopathies----structural abnormalities
in polypeptide chain
i. Hemoglobin S
It is found in sickle cell anemia (abnormal Beta chain)
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ii. Hb C------The β-chains are abnormal,
Glutamic acid replaced by Lysine at position
no.6, which is characterized by mild hemolytic
anemia and splenomegaly.
iii. Hb E-------The β-chains are abnormal, Glutamic
acid replaced by Lysine at position no.26

 Hb in Thalassemia----polypeptide chains are

decreased, absent or abnormal.
1. In α-thalassemia, the α-chains are absent or dec.
2. In β-thalassemia, the β-chains are absent or dec.
 Destruction of Hb
• 120 days of RBCs-----destroyed by RES in
spleen.
• Hb is degraded into globin, iron and porphyrin
• IRON---stored in body as ferritin &
hemosiderin-------reutilized for Hb synthesis
• Globin--utilized for resynthesis.
• Porphyrin---It is converted into biliverdin
which is converted into bilirubin.
• Biliverdin & bilirubin are called bile pigments
ERYTHROPOEISIS
 Areas of Body that Produce RBCs
• In the early weeks of embryonic life,
primitive, nucleated red blood cells are
produced in ----yolk sac.
• Middle trimester of gestation------liver is
main organ but RBCs also produced in spleen
& lymph nodes.
• During the last month of gestation and after
birth--------red blood cells are produced in
BONE MARROW.
• UPTO 5 years: Bone marrow of all bones
produces RBCs.

• 5-20 years: MARROW OF LONG BONES

except humerus and tibia

• After age 20 years: marrow of the

membranous bones (vertebrae, sternum, ribs,
scapulas, and iliac bones)
 Stages of Differentiation of Red
Blood Cells
• Proerythroblast.
• Under appropriate stimulation, large numbers
of these cells are formed from the CFU-E stem
cells.
• It divides multiple times forming many mature
red blood cells
• The first-generation cells are called basophil erythroblasts
because they stain with basic dyes; the cell has very little
hemoglobin.
• Endoplasmic reticulum is reabsorbed, consisting of
remnants of the Golgi apparatus, mitochondria, and a few
other cytoplasmic organelles.

• During this reticulocyte stage, the cells pass from the

bone marrow into the blood capillaries by diapedesis
(squeezing through the pores of the capillary membrane)

• The remaining basophilic material in the reticulocyte

normally disappears within 1 to 2 days, and the cell is
then a mature erythrocyte.
Regulation of Red Blood Cell Production-
Role of Erythropoietin
• RBCs are regulated
(1) adequate red cells are always available to
provide sufficient transport of oxygen from the
lungs to the tissues

(2) the cells do not become so numerous that

they impede blood flow.