PHM142 Lecture 4: Platelets + Endothelial Cells

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1 PHM142 Lecture 4: Platelets + Endothelial Cells 1

2 Hematopoiesis 2

3 Platelets Critical in clotting - activated by subendothelial matrix proteins (e.g. collagen, fibronectin, von Willebrand factor) and thrombin Half life: 4-10 days Contain: glycogen granules, mitochondria, lysosomes Do not contain: nucleus, DNA, protein synthesis Thrombopoietin: increases platelet numbers 3

4 Blood Clotting: Overview 4

5 1. Vascular Phase initial: neurogenic and myogenic control prolonged by: serotonin, thromboxane A2 (platelets) endothelin-1 (endothelial cells) 5

6 2. Platelet Phase Exposed collagen -> platelet binding (via von Willebrand Factor) + platelet activation Change in platelet morphology Release of platelet factors -> platelet aggregation -> vasoconstriction alpha granules: fibrinogen, von Willebrand Factor, PDGF dense granules: serotonin, calcium, ADP, ATP platelet activating factor (PAF), thromboxane A2 6

7 Endothelium in Clotting Normally: Prostaglandins and Nitric Oxide (NO) prevent platelet adhesion and promote vasodilation 7

8 3. Coagulation Phase Two Upstream Pathways: 1. Intrinsic (Contact Activation) Pathway: Stimulated by: Collagen, Plasma proteins 2. Extrinsic (Tissue Factor) Pathway Stimulated by: Tissue thromboplastin Converge on the Common Pathway 8

9 Common Pathway 9

10 Intrinsic Pathway von Willebrand Factor regulates VIII Vitamin K required for thrombin, VII, IX, and X synthesis Warfarin (Coumadin) inhibits Vitamin K activity 10

11 Vitamin K vitamin K warfarin Vitamin K is required for synthesis of prothrombin and clotting factors VII, IX, and X warfarin (Coumadin) is an oral anticoagulant that acts as a vitamin K antagonist therefore patients on warfarin should not exceed a dietary intake of micrograms of Vitamin K per day 11

12 Extrinsic Pathway 12

13 Platelets: the Big Picture Repair of Damaged Vascular Walls: Platelet Derived Growth Factor (PDGF) -> migration + division of vascular endothelial cells, smooth muscle cells, fibroblasts Clot Retraction: Membrane Actin and Myosin contract -> fibrinattached platelets retract -> broken blood vessel edges pulled together 13

14 Platelet Derived Growth Factor PDGF-A, PDGF-B, PDGF-C, PDGF-D disulfide-linked dimers growth factor core domain: conserved cysteines 14

15 PDGF Processing N-term pro-domain of PDGF-A (70 aa) and likely PDGF-B (65 aa) removed by furin N-term prodomain of PDGF-C (212 aa) removed by plasmin and tissue plasminogen activator (tpa) N-term prodomain of PDGF-D (239 aa) removed by plasmin 15

16 Fibrinolysis tpa 16

17 Bleeding + Clotting Disorders Haemophilia von Willebrand Disease Thrombocytopenia 17

18 Prostanoids Family of lipid mediators Common precursor: Arachidonic acid Prostanoids: Prostaglandins PGD2, PGE2, PGF2α, PGI2 (prostacyclin), and TXA2 Prostaglandin Synthase 18

19 Membrane phospholipid Structure: glycerol backbone fatty acid at the first position arachidonic acid at the second position polar phospholipid moiety (Choline/ Serine/ Inositol/ Ethanolamine) at the third position 19

20 Prostanoid Biosynthesis Arachidonate (released by phospholipases) PGG2 PGH2 PGI2 PGE2 Thromboxane A2 PGD2 PGF2 20

21 Thromboxane A 2 + Aspirin acetylation of Ser530 TxA 2 synthase TxA 2 : a potent vasoconstrictor involved in platelet recruitment + activation acetylsalycylic acid irreversibly acetylated and inhibits COX thereby blocking TxA 2 production side effects: GI toxicity as a result of decreased PGE2 levels + acidosis 21

22 Endothelial Cells Nature s container for blood Functions: inflammation, wound healing, fibrosis, smooth muscle cell proliferation, angiogenesis, and blood pressure Balance between dilation (NO) and constriction (angiotensin) 22

23 Nitric Oxide 1992: Molecule of the Year Endothelium -> Nitric Oxide -> cgmp -> smooth muscle relaxation -> vasodilation -> increased blood flow Synthesized from L-Arginine by enos Drugs: Nitroglycerin Isopentyl Nitrite 23

24 Angiotensin Angiotensin converting enzyme (ACE): present on endothelial cell surface ACE inhibitors 24

25 Maya Latif - maya.latif@mail.utoronto.ca TAs: Ersa Gjelaj - ersa.gjelaj@mail.utoronto.ca Andrew Fleet - andrew.fleet@mail.utoronto.ca Office hours: Tues 11-12, Thurs 12-1, room 900/1000 MIDTERM: Friday October 19th, 11:45-2:45, EX100

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