Beruflich Dokumente
Kultur Dokumente
BY DR komal makwana 2nd YEAR RESIDENT , DEPARTMENT OF PHYSIOLOGY, MEDICAL COLLEGE BHAVNAGAR.
OUTLINE
History Formation Physical and Chemical Characteristics of Platelets Functions- Platelet Plug Formation Drug developed &Applied physiology(not as separate part ,it will walk along with physiology.)
HISTORY
ADDISON in 1841 described platelets as extremely minute granules in clotting blood. They were termed platelets by BIZZOZERO who observed their adhesive properties. They were microscopically examined by OSLER & SCHAEFER and by HAYEM in 19th century. In 1906 JAMES HOMER WRIGHT put the hypothesis that platelets are derived from the cytoplasm of the megakaryocytes and basis of thrombopoieses were established.
Formation of platelets
Megakayocytes
Platelets arise from megakaryocytes Largest cells in the body (30-50um) Polyploid Cluster on extravascular compartment The megakaryocytes, giant cells in the bone marrow, form platelets by pinching off bits of cytoplasm and extruding them into the circulation
Platelets (also called thrombocytes) are minute discs 1 to 4 micrometers in diameter.(Greek word thrombus- plates. ) The normal concentration of platelets in the blood is between 150,000 and 4,00,000 per micro litre.
Platelets are emerging as remarkable cell fragment with abundant metabolic capability but minimal ability to synthesize protein because it contains only law levels of RNA and lacks nucleas. Stress platelets- are large and beaded. Reticulated platelets- large, high RNA, recently released from the marrow.
Platelet surface
Platelet plasma membrane Glycocalyx Membrane rafts or GEMS-Glycolipid enriched membrane domain Surface connected canalicular system
Glycocalyx 20-30 nm Thicker than WBCs and RBC. Platelets carries its functional environment with it to maintain a negative environment Anchored within the membrane are: 1. Glycoprotein 2. Proteoglycans Endocytosis: transport mechanism used by the glycocalyx
Route for granule release during platelet activation and secretion. Route for ingress and egress of molecules as they translocate between plasma and platelet.
Storage organs
Four types granules Dense bodies Lysosomes microperoxisomes
granules
Electron dense zones
less Electron density zone
granules also have fibrinogen which is actively incorporated from plasma. PDGF,VEGF(angiogenesis stimulator),endostatin(angiogenesis inhibitor)
Dense bodies
3 to 5/cell Bulls eye appearance electron dense ATP , pyrophosphate, calcium, serotonin, GTP, GDP, magnesium.
Platelet Cytoskeleton
Microfilaments and Microtubules In general functions for: 1. platelet shape changes 2. extension of pseudopods 3. secretion of granule contents
Platelet Ultrastructure
Platelet Cytoskeleton: Mricrofilaments and Microtubules
Microfilaments are thick meshwork of actin filaments Actin is contractile and anchors the membrane glycoproteins and proteoglycans Actin also present throughout the cytoplasm, making 20-30% of platelet proteins
Platelet Ultrastructure
Platelet Cytoskeleton: Microfilaments and Microtubules
Intermediate Filaments: ropelike polymers; 8-12 nm in diameter; made up of desmin and vimectin Connects with actin and the microtublues and helps maintain the cell shape
Platelet function:
A)Platelet function:
i. Blood clotting and stroke (anticoagulant drugs) ii. Prostaglandin and thromboxane formation from polyunsaturated fatty acids iii. Nutritional approach to prevent heart disease iv. COX Inhibitors (NSAIDs) (antiplatelet drugs)
First we will see the events of hemostasis superficially, After getting the idea of major events we will study the role of platelet in detail.
GPIa/IIa and GPIb are platelet membrane proteins that bind to collagen and von Willebrand factor (vWF), causing platelets to adhere to the subendothelium of a damaged blood vessel. PAR1 and PAR4 are protease-activated receptors that respond to thrombin (IIa); P2Y1 and P2Y12 are receptors for ADP (adenosine diphosphate); when stimulated by agonists, these receptors activate the fibrinogen-binding protein GPIIb/IIIa and cyclooxygenase-1 (COX-1) to promote platelet aggregation and secretion. Thromboxane A2 (TXA2) is the major product of COX-1 involved in platelet activation. Prostaglandin I2 (PGI2), synthesized by endothelial cells, inhibits platelet activation.
interactions among proteins of the "extrinsic" (tissue factor and factor VII), "intrinsic" (factors IX and VIII), and "common" (factors X, V, and II) coagulation pathways Factor xa bind to factor 5a on the surface of activated platelet , a specific surface protein to which Factor X can bind, which is missing in scott syndrome.
2. Immunglobulin Gene GP VI: binds collagen and adhesive protein Thrombospondin 3. Leucine-rich Repeat Family GP Ib/IX/V: 7 non covalently bound molecules with ratios 2:2:2:1(will see them in figure) (2) GP 1ba: accounts for VWF binding; binding of thrombin (2) GP IbB: crosses the membrane and interacts with actin binding protein: outside in signalling(we will see what is the outside in signaling)
3. Leucine-rich Repeat Family (2) GP IX: associated with mucocutaneous bleeding disorder and Bernard Soulier Syndrome (1) GP V
VWF Function
Adhesion Mediates the adhesion of platelets to sites of vascular injury (subendothelium) Links exposed collagen to platelets Mediates platelet to platelet interaction Binds GPIb and GPIIb-IIIa on activated platelets Stabilizes the hemostatic plug against shear force
vWF VWF gene : short arm of chromosome 12 VWF gene is expressed in endothelial cells and megakaryocytes vWF is produced as a propeptide which is extensively modified to produce mature vWF Two vWF monomers bind through disulfide bonds to form dimers Multiple dimers combine to form vWF multimers
Vwf has A1 and A3 domains that binds to collagen. The A1 domain contains binding site for GP1b complex, primary role player in adhesion, changes its conformation in response to high shear forces, thus making high affinity ligand for GPIb complex.
ITAM lead to recruitment of the tyrosine kinase (Syk) through its tandem Src-homology(SH2) So activation of syk occurs, which lead to phosphorylation of phospholipase C 2(PLC 2 C 2). PLC 2 play critical role in aggregation and secretion- generation of second messengers ITP & DAG- intracellular calcium rises & activation of PKC.
Why arteries have white clot? And veins have red? At high shear rate, arteries gpIb/v/IX receptors and vwf ligand play major role, fibrinogen is only stabilizing factor. Low shear rates , veins fibrinogen IIb 3 supporting platelet plug formation, produce red clot.
THROMBIN comes and cleave the peptide bond between Arginine 41 and Serine42. These tethered ligand join irreversibly with the body of the receptor gives transmembrane signaling. subunits of G12,G13 platelet shape change subunits of Gq platelet secretion aggregation ,participates in activation of PIC PIC - calcium mobilization and PKC activation.
Thrombin ADP
TxA2
GP IIb/IIIa
Phospholipase C
Ca2+ Influx
61
SIGNAL srC- syk(tyrosin kinase) that bound to tail of B3- adaptor protein SLP 76 Rac GTPase vav.- all these lead to actin reorganization.
Platelet Activation-Secretion
utside in platelet activation through ligand binding to Integrins and Seven transmembrane receptors (STRs) Triggers actin microfilament contraction Intermediate filaments and microtubules contract = compression of granules Contents of the alpha granules and lysosomes flow through the SCCS Contents of delta granules are secreted through the plasma membrane
Platelet in atherogenesis
ANTIPLATELET DRUGS
Platelets provide the initial hemostatic plug at sites of vascular injury, participate in pathological thromboses that lead to myocardial infarction, stroke, and peripheral vascular thromboses. Potent inhibitors of platelet function have been developed in recent years.
These drugs act by discrete mechanisms, and thus in combination their effects are additive or even synergistic. Their availability has led to a revolution in cardiovascular medicine, whereby angioplasty and vascular stenting of lesions now is feasible with low rates of restenosis and thrombosis when effective platelet inhibition is employed
In platelets, the major cyclooxygenase product is thromboxane A2, a labile inducer of platelet aggregation and a potent vasoconstrictor. Aspirin blocks production of thromboxane A2 by acetylating a serine residue near the active site of platelet cyclooxygenase (COX-1), the enzyme that produces the cyclic endoperoxide precursor of thromboxane A2.
Since platelets do not synthesize new proteins, the action of aspirin on platelet cyclooxygenase is permanent, lasting for the life of the platelet (7 to 10 days). repeated doses of aspirin produce a cumulative effect on platelet function. Complete inactivation of platelet COX-1 is achieved when 160 mg of aspirin is taken daily
Dipyridamole interferes with platelet function by increasing the cellular concentration of adenosine 3,5-monophosphate (cyclic AMP). This effect is mediated by inhibition of cyclic nucleotide phosphodiesterase and/or by blockade of uptake of adenosine, which acts at adenosine A2 receptors to stimulate platelet adenylyl cyclase. The only current recommended use of dipyridamole is in combination with warfarin for postoperative primary prophylaxis of thromboemboli in patients with prosthetic heart valves.
Ticlopidine (TICLID) is a that inhibits the P2Y12 receptor. Ticlopidine is a prodrug that requires conversion to the active thiol metabolite by a hepatic cytochrome P450 enzyme. It permanently inhibits the P2Y12 receptor by forming a disulfide bridge between the thiol on the drug and a free cysteine residue in the extracellular region of the receptor and thus has a prolonged effect
Abciximab. (REOPRO) is the Fab fragment of a humanized monoclonal antibody directed against the aIIbb3 receptor. It also binds to the vitronectin receptor on platelets, vascular endothelial cells, and smooth muscle cells. The antibody is used in conjunction with percutaneous angioplasty for coronary thromboses, and when used in conjunction with aspirin and heparin, has been shown to be quite effective in preventing restenosis, recurrent myocardial infarction, and death.
Eptifibatide (INTEGRILIN) is a cyclic peptide inhibitor of the fibrinogen binding site on aIIbb3. It blocks platelet aggregation. It is used to treat acute coronary syndrome and for angioplastic coronary interventions. its benefit is somewhat less than that obtained with the antibody Abciximab , perhaps because eptifibatide is specific for aIIbb3 and does not react with the vitronectin receptor.
Platelet disorder
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