INFLAMMATION 2

CONTENTS
• Chemical mediators of inflammation
• General principals and classification of chemical mediators
LEARNING OBJECTIVES
• Be able to define and classify the chemical mediators of inflammation
MEDIATORS OF INFLAMMATION

1. Substances that initiate & regulate inflammatory reactions.

2. The most important mediators of acute inflammation are vasoactive amines,
lipid products (Prostaglandins & leukotrienes), cytokines (Chemokines) & products
of complement activation
GENERAL PRINCIPLES
A. Mediators may be produced locally by cells at the site of inflammation
ii. May be derived from circulating inactive precursors that are activated at the
site of inflammation.
iii. Cell-derived mediators are rapidly released from IC granules (Amines) or
synthesized de novo (Prostaglandins, leukotrienes, cytokines) in response to a
stimulus.
iv. The major cell types that produce mediators of acute inflammation are
tissue macrophages, dendritic cells & mast cells, but platelets, neutrophils,
endothelial cells & most epithelia also can be induced to elaborate mediators.
v. Cell-derived mediators are most important for reactions against offending
agents in tissues.
vi. Plasma-derived mediators (Complement proteins) are present in the
circulation as inactive precursors that must be activated. They are produced
mainly in the liver.
B. Active mediators are produced only in response to various molecules that
stimulate inflammation
viii. Many of these stimuli trigger well defined receptors & signaling pathways
C. Most mediators are short-lived, quickly decay, inactivated by
enzymes, scavenged or inhibited.
System of checks & balances regulates mediator actions.
D. One mediator can stimulate the release of other mediators e.g.
products of complement activation stimulate the release of histamine
& cytokine TNF acts on endothelial cells to stimulate the production of
another cytokine---IL-1 & many chemokines.
The secondary mediators may have the same actions as the initial
mediators or different & even opposing activities.
PRINCIPAL MEDIATORS OF
INFLAMMATION
Mediator Source Action
1. Histamine, Mast cells Vasodilation
Basophils Increased vascular permeability Endothelial activation
Platelets

1. Prostaglandins Mast cells Vasodilation

Leukocytes Pain
Fever

1. Leukotrienes Mast cells Increased vascular permeability, chemotaxis

Leukocytes Leukocyte adhesion/activation

1. Cytokines (TNF, IL-1, IL-6) Macrophages Local: Endothelial activation (Expression of adhesion molecules). Systemic:
Endothelial cells Fever, metabolic abnormalities, hypotension (shock)
Mast cells

1. Chemokines Leukocytes Chemotaxis

Activated macrophages Leukocyte activation

1. Platelet-activating factor Leukocytes Vasodilation

Mast cells Increased vascular permeability, Leukocyte adhesion
Chemotaxis
Degranulation
Oxidative burst

1. Complement Plasma (Liver) Leukocyte chemotaxis and activation, direct target killing (membrane attack
complex), vasodilation (mast cell stimulation)

1. Kinins Plasma (Liver) Increased vascular permeability, smooth muscle contraction, vasodilation, pain
1. Vasoactive Amines
i. Histamine & Serotonin: Have important actions on blood vessels
ii. Stored as preformed molecules in cells
iii. Among the first mediators to be released during inflammation.
iv. The richest sources of histamine are mast cells normally located
adjacent to blood vessels.
v. Histamine is also found in blood basophils & platelets.
vi. It is stored in mast cell granules, released by degranulation in
response to a variety of stimuli:
a. Physical injury (Trauma, cold, or heat, etc.
b. Binding of antibodies to mast cells, which underlies immediate
hypersensitivity (allergic) reactions
c. Products of complement (Anaphylatoxins--C3a & C5a)
d. Antibodies & complement products bind to specific receptors on mast
cells & trigger signaling pathways that induce rapid degranulation.
e. Neuropeptides (Substance P) & cytokines (IL-1, IL-8) may trigger
release of histamine.
Histamine causes dilation of arterioles & increases the permeability of
venules.
i. Principal mediator of the immediate transient phase of increased vascular
permeability, producing interendothelial gaps in postcapillary venules.
ii. Its vasoactive effects are mediated mainly via binding to receptors, called H1
receptors, on microvascular endothelial cells.
iii. Antihistamine commonly used to treat inflammatory reactions e.g. allergies
are H1 receptor antagonists that bind to & block the receptor.
iv. Histamine also causes contraction of some smooth muscles
• Serotonin (5-hydroxytryptamine): Is a preformed vasoactive mediator
present in platelets & certain neuroendocrine cells (GIT)
• Its primary function is as a neurotransmitter in the GIT.
• It is also a vasoconstrictor
Arachidonic Acid Metabolites
• The lipid mediators (Prostaglandins & leukotrienes) are produced from arachidonic acid (AA) present in
membrane phospholipids
• They stimulate vascular & cellular reactions in acute inflammation. AA: Polyunsaturated fatty acid derived
from dietary sources or by conversion of the essential fatty acid (linoleic acid).
• Most cellular AA is esterified & incorporated into membrane phospholipids.
• Mechanical, chemical, physical stimuli or other mediators trigger the release of AA from membranes by
activating cellular phospholipases, mainly phospholipase A2.
• Once released from the membrane, it is rapidly converted to bioactive mediators called eicosanoids which
are synthesized by 2 major classes of enzymes:
• Cyclooxygenases (Generate prostaglandins)
• lipoxygenases (Generates leukotrienes & lipoxins).
• Eicosanoids bind to G protein-coupled receptors on many cell types & can mediate virtually every step of
inflammation
•
PROSTAGLANDINS
• Prostaglandins (PGs): Produced by mast cells, macrophages, endothelial
cells are involved in vascular & systemic reactions of inflammation. They
are generated by the actions of 2 cyclooxygenases (COX-1 & COX-2)
• COX-1 is produced in response to inflammatory stimuli & also serve a
homeostatic function (Fluid & electrolyte balance in the kidneys,
cytoprotection in the GIT).
• In contrast, COX-2 is induced by inflammatory stimuli & thus generates
the PGs that are involved in inflammatory reactions, but it is low or
absent in most normal tissues.
• The most important prostaglandins in inflammation are PGE2, PGD2,
PGF2a, PGI2 (Prostacyclin) & TXA2 (Thromboxane A2)
• PGD2 is the major prostaglandin made by mast cells; along with PGE2
(More widely distributed), it causes vasodilation & increases the
permeability of postcapillary venules, thus potentiating exudation &
resultant edema.
• PGD2 also is a chemoattractant for neutrophils.
• Platelets contain the enzyme Thromboxane synthase responsible for
synthesizing TXA2 (Major platelet eicosanoid). TXA2 is a potent
platelet-aggregating agent & vasoconstrictor, promoting thrombosis.
In contrast, vascular endothelium contains prostacyclin synthase,
responsible for the formation of prostacyclin (PGI2) & its stable end
product PGF1a.
Prostacyclin is a vasodilator & a potent inhibitor of platelet aggregation,
thus serves to prevent thrombus formation on normal endothelial cells.
Thromboxane– prostacyclin imbalance has been implicated in coronary
& cerebral artery thrombosis
• Prostaglandins are involved in the pathogenesis of pain & fever
LEUKOTRIENES
Leukotrienes produced in leukocytes & mast cells by the action of lipoxygenase
Involved in vascular & smooth muscle reactions
Leukocyte recruitment.
The synthesis of leukotrienes involves multiple steps:
• First, generates leukotriene A4 (LTA4), which in turn gives rise to LTB4 or LTC4.
• LTB4 is produced by neutrophils & macrophages
• Potent chemotactic agent & activator of neutrophils, causing aggregation & adhesion of the cells
to venular endothelium
• Generation of ROS & release of lysosomal enzymes.
• The cysteinyl-containing leukotriene LTC4 & its metabolites, (LTD4 & LTE4), are produced mainly in
mast cells & cause intense vasoconstriction, bronchospasm (Asthma) & increased permeability of
venules.
LIPOXINS
Generated from AA by the lipoxygenase pathway
Unlike prostaglandins & leukotrienes, lipoxins suppress inflammation by
inhibiting the recruitment of leukocytes.
They inhibit neutrophil chemotaxis & adhesion to endothelium.
Two cell populations are required for the transcellular biosynthesis of
these mediators.
Neutrophils produce intermediates in lipoxin synthesis which are
converted to lipoxins by platelets interacting with the leukocytes.
Inhibitors of Prostaglandins & Leukotrienes
Importance of eicosanoids in inflammation: Drugs that inhibit their
production or actions, suppressing inflammation (Anti-inflammatory)
Cyclooxygenase inhibitors: Aspirin, non-steroidal anti-inflammatories
(NSAIDs) e.g. Ibuprofen. They inhibit both COX-1 & COX-2 blocking all
prostaglandin synthesis (Pain & fever)
Aspirin by irreversibly inactivating cyclooxygenases.
Selective COX-2 inhibitors are X200-300 more potent in blocking COX-
2 than COX-1.
COX-1 is responsible for the production of prostaglandins involved in
both inflammation & physiologic functions
COX-2 generates prostaglandins involved only in inflammation
Selective COX-2 inhibitors may increase the risk of CV & cerebrovascular
events via impairing of endothelial cell production of prostacyclin
(PGI2), which prevents thrombosis, while leaving intact the COX-1-
mediated production by platelets of TXA2, which induces platelet
aggregation
Lipoxygenase inhibitors
1. 5-lipoxygenase is not affected by NSAIDs
2. Corticosteroids: Broad-spectrum anti-inflammatory that reduce the
transcription of genes encoding COX-2, phospholipase A2,
proinflammatory cytokines (e.g., IL-1 and TNF) & iNOS.
3. Leukotriene receptor antagonists block leukotriene receptors and
prevent the actions of the leukotrienes, useful in the treatment of
asthma.
Cytokines & Chemokines
1. Cytokines are proteins secreted by many cell types (Principally
activated lymphocytes, macrophages &dendritic cells, but also
endothelial, epithelial & CT cells) that mediate & regulate immune
& inflammatory reactions.
2. By convention, GFs that act on epithelial & mesenchymal cells are
not grouped under cytokines.
Cytokines involved in acute inflammation
Tumor Necrosis Factor & Interleukin-1
1. TNF & IL-1 serve critical roles in leukocyte recruitment by promoting adhesion of
leukocytes to endothelium & migration through vessels.
2. Activated macrophages & dendritic cells mainly produce these cytokines TNF also
is produced by T lymphocytes & mast cells & some epithelial cells produce IL-1.
3. Microbial products, foreign bodies, necrotic cells & other inflammatory stimuli
can stimulate the secretion of TNF & IL-1.
4. The production of TNF is induced by signals through TLRs & other microbial
sensors & the synthesis of IL-1 is stimulated by the same signals, but the
generation of the biologically active form of this cytokine is dependent on the
inflammasome.