Platlets – thrombocytes

2.5 um, thickness 0.5-1 um' v = 8 fl, life span 8-12 days
count: 100 – 300,000/ul, 300 x 10^9/l, 3 x 10^11/l
2/3 is in blood, 1/3 in spleen
without sex and age dependence
change in ovulation, menstruation
pregnancy – decrease
stress, exercise, adrenaline – increase

Functions
prevention of blood lose (by mechanical and humoral effects)
healing of the wounds: PDGF (platlets derived growth factor)
change of the mambrane permability
participation at inflamation
participation at transplatation
transport of substances

Composition of Tr

thrombocytes =
1. dense granules --> serotonin, ADP, ATP, Ca^+2
2. (alpha) – granules --> platelets factors 1,2,5. PDGF, vWF, XIII, PAF, factor 4 – antiheparin
3. lysosomes
4. tubules: Ca^+2, enzymes for arachidonic acid metabolism.
5. Phospholipids: platlets factor 3, source of arachidonic acid: tromboxane A2, prostacyclin,
prostaglandin, PAF, leucotriens.
6. Contractile apparatus: actin, myosin
7. rectractoenzyme
8. receptors (glycoproteins)
9. antigens (glycoproteins) PI^A, PI^E, ABO, HLA ---> antibodies: after transfusion, gravidity,
transplantation.

Hemostasis

Defense mechanism, complex of processes stopping of bleeding.

Prevention of blood loss
formation of a clot
homeostatic mechanism

Mechanisms of hemostasis

1. Local vasoconstriction
2. formation of platlet plug
3. formation of a blood clot (definite clot)

Injury

1. vasoconstriction
 2. temporary plug – white thrombus
3. blood clotting – clot retraction – fibrinolysis

1. Local vasoconstriction
– local myogenic spasm (to injury of vessels immediate, myogenic reaction)
– reflex spasm: reaction to the pain, exeroreceptors – efferent pathway – sympathetic fibers
– humoral vasoconstriction: serotonin, adrenaline, fibrinopeptides

depend on: kind of injury, extension of bleeding, speed of bleeding (arterial, venous, capillary).

2. formation of platelet plug (provisional sealing)

– adhesion: due to collagen fibers

von willebrand factor (VIII)
trombin
ADP
– metamorphosis of Pl
– degranulation (due to actin and myosin) – substances granules are released to the blood and
injury zone.
– Aggregation: ADP, tromboxan A2, platlet activating factor (PAF), serotonin, adrenaline,
thrombin, fibrinogen, vWF (VIII)

Ca^+2 in TC rises, cAMP decreases

reversible aggregation = thrombocyte plug = white thrombus

3. Blood coagulation

Moravitz, 1900, 4 phases

1. thrombokinase formation
prothrombin – thrombin

2. fibrinogen – fibrin

3. retraction of fibrin

4. fibrinolysis

1960 – congulation cascade, enzymatic processes, serinorotease

factors needed for coagulation: I – XIII
HMW-K (high molec. Weight, kininogen, prekallikrein, Pl, Le.

Clotting factors in the blood

1. Fibronogen
2. Prothormbin
3. Tissue factor
 4. Calcium
5. Proaccelerin, labile factor
6. proconvertin, stable factor
7. antihemophilic factor A
8. plasma thromboplastin component
Christmas factor, antihemophilic fact
9. Stuart factor, Stuart-prower factor, Plasma

Intrinsic pathway
extrinsic pathway
common pathway

Injury image – tissue, endothelium

Regulation of hemostasis

hemostasis -- * ----> inhibition of clotting

Vasconstriction intact endothelium – prevent adhesion, prevent aggregation
blood flow – dilution of clotting factors
platelet plug prostacyclin – vasodilatation
EDRF – endothelium derived relaxing factor (NO)
clotting: intrinsic pathway fibrin threads (antitrombin)
exrinsic “ antitrombin III, cofactor of heparin
antiheparin (pl. f 4) plasminogen – plasmin
antiplasmine plasminogen activators

intravascular coagulation hemorrhage

thrombus, embolus

Fibrinolysis

plasminogen: (increase) infection, injury, surgery intervention, intract IM

after activation --> plasmin <-- antiplasmin
plasmin splits fibrin, fibrinogen, prothrombin, V, VIII

inhibition

activation
1. intrinsic system:
venostasis
XIIA, katecolamines,
kallikrein, thrombin
2. extrinsic system: intima and tissue
proteases = tissue activators
after injury – the most important
3. exogenous factors: urokinase, streptokinase
human tissue activator of plasmin
- clinical utilisation: infract-IM
Fibrinolytic system

daily rythm:

during day increases

during night decreases
increase – physical activity, stress, during menstruation, pregnancy (female)

Anticoagulants

substances that prevent blood clotting

in vitro: (sampling)
– defibrination
– contact with nonwettable surface of test tubes (silicone, paraffin)
– calcium salts inactivation
sodium citrate 3.8% (1:4), precipitates Ca^+2
sodium oxalate (1:9)
chelate – Na2EDTA (ethylene diamine tetraacetic acid)
– heparin (natural anticoagulants)
– tempratures decreasing

In vivo: treatment, prevention

– heparin – natural anticoag. (inhibits active factors IX, X, XI, XII)
– coumarin derivatives – dicumarol – of plant origin
inhibits vitamin K uptake by the liver factor: II, VII, IX, X
– soya beans (in food)
– aspirin
– liver disseases
– vit. K deficiency
– hirudin (leech, insects) prevents action of trombin
– poison of snakes...

Acceleration of coagulation

in vitro:
– foreign bodies
– temperature increasing
– water wettable surface

in vivo:
– roughning of the endothelial lining
– blood stagnation
– injury
– extensive tissue damage
– hypertonic solution (i.v)
– vitamin K, adrenaline
 – calcium

Investigation:

– bleeding time: 1-4 min.

– clotting time: 3-8 min (on watch glass)
– Quick prothrombin time: 12-14 sec., 75-100%
test of the extrinsic pathway: VII, X, V, II, I

– APTT (activated partial thromboplastin time)

test of intrinsic and common pathway: 35-45s, 80-100%: XIII, IX, XI, X, V, II, I
– Lee white test – clotting time (in test tubes, T:37C
– Rumpel Leed: test of resistance of blood capillaries

blood plasma
blood serum: without I, II, VIII, XIII

hemophilia: bleeding time & prothrombin time are normal, clotting time is prolonged.