Session 9: Chemical mediators

of Inflammation

1
 Learning tasks
At the end of this session, students are expected to be
able to:
• Define chemical mediators of inflammation.
• Identify general properties of chemical mediators of
inflammation.
• Identify different types of chemical mediators of
inflammation.
• Explain mechanisms of action of different types of
chemical mediators of inflammation.

2
 Definition
• Chemical mediators of inflammation can be
defined as any soluble substance that act on the
blood vessel, inflammatory cells or any other
cell to contribute to an inflammatory response.
Different chemical mediators
 General properties of chemical
mediators of Inflammation
1. Mediators may be produced locally by cells at
the site of inflammation or may be derived from
circulating inactive precursors (typically
synthesized by the liver).
2. Most mediators act by binding to specific
receptors on different target cells.
3. The actions of most mediators are tightly
regulated and short lived.
4. Mediators may stimulate target cells to release
secondary effector molecules.
 Properties of chemical mediators of
Inflammation
1. Mechanism of action by:
– Receptor-ligand interactions (1o).
– Direct enzymatic activity.
– Mediate oxidative damage.
2. Extensive network of interacting chemicals.
3. High degree of redundancy.
4. Guarantees amplification and maintenance of
inflammatory response.

6
 Properties of chemical mediators of
Inflammation cont…
5. Short half life and are harmful.
6. Active mediators are produced in response to
various stimuli.
7. One mediator can stimulate the release of other
mediator.
8. If unchecked, mediators can cause harm.

7
 Types of chemical mediators of
Inflammation
According to their source:
1. Cellular derived
2. Plasma derived
• OR can be derived:
1. Exogenous
• Endotoxins
2. Endogenous
•Plasma
•Leukocytes
•Endothelial cells
•Fibroblasts
 Cellular derived chemical
mediators of inflammation
• Cells which produce chemical mediators of
inflammation are:
– Tissue macrophages, mast cells and
endothelial cells at the site of inflammation.
– Leukocytes that are recruited to the site from
the blood.
 Cellular derived chemical
mediators of inflammation cont…
1. Vasoactive amines:
– Histamine.
– Serotonin.
2. Arachidonic acid metabolites:
– Prostaglandins.
– Thromboxanes.
– Leukotrienes.
– Lipoxins.
 Cellular derived chemical
mediators of inflammation cont…
3. Platelet Activating Factor (PAF).
4. Cytokines:
– Tumor Necrosis Factor (TNF).
– Interleukin 1 (IL-1).
– Interleukin 6 (1L-6).
– Chemokines.
5. Reactive oxygen species (ROS).
 Cellular derived chemical
mediators of inflammation cont…
6. Nitric oxide (NO).
7. Lysosomal enzymes:
– Neutral proteases.
8. Neuropeptides:
– Substance P.
 Vasoactive Amines
• The two vasoactive amines are:
i. Histamine.
ii. Serotonin.
• These are stored as preformed molecules in
mast cells and other cells (like serotonin and
lysosomal enzymes are also preformed mediators).
• These are among the first mediators to be
released in acute inflammatory reactions.
 Histamine
• Histamine is produced by many cell types:
– Mast cells adjacent to vessels.
– Circulating basophils.
– Platelets.
• Histamine causes:
i. Arteriolar dilation.
ii. Increases vascular permeability.
iii. Endothelial activation.
• Soon after its release, histamine is inactivated by
histaminase.
 Histamine cont…
• Preformed histamine is released from mast cell
granules in response to a variety of stimuli:
1. Physical injury such as trauma or heat.
2. Immune reactions involving binding of IgE
antibodies to Fc receptors on mast cells.
3. C3a and C5a fragments of complement, the so-
called anaphylatoxins.
4. Leukocyte-derived histamine-releasing proteins.
5. Neuropeptides (e.g., substance P).
6. Certain cytokines (e.g., Interleukins 1 and 8).
 Serotonin
• Serotonin (5-hydroxytryptamine) is a preformed
vasoactive mediator found within platelet
granules.
– It is released during platelet aggregation.
– It induces vasoconstriction during clotting.
• It is produced mainly in some neurons and
enterochromaffin cells.
• It is a neurotransmitter and regulates intestinal
motility.
• Causes vasodilation and increase vascular
permeability.
Arachidonic Acid (AA) metabolites
• AA metabolites, also called eicosanoids (because
they are derived from 20-carbon fatty acids—
Greek eicosa, “twenty”), can mediate virtually
every step of inflammation.
• Their synthesis is increased at sites of
inflammatory response and agents that inhibit their
synthesis also diminish inflammation.
Arachidonic Acid (AA) metabolites cont…
• AA is a 20-carbon polyunsaturated fatty acid (with
four double bonds) produced primarily from
dietary linoleic acid.
– Present in the body mainly in its esterified form as a
component of cell membrane phospholipids.
• It is released from these phospholipids through the
action of cellular phospholipases.
– Phospholipases are activated by mechanical, chemical,
or physical stimuli, or by inflammatory mediators such
as C5a.
Arachidonic Acid (AA) metabolites cont…
• AA metabolism proceeds along one of two major
enzymatic pathways:
i. Cyclooxygenase pathway stimulates the synthesis
of prostaglandins and thromboxanes.
ii. Lipoxygenase pathway is responsible for
production of leukotrienes and lipoxins.
• Thus AA metabolites are:
1. Prostaglandins.
2. Thromboxanes.
3. Leukotrienes.
4. Lipoxins.
Arachidonic Acid (AA) metabolites
cont…
• Sources of AA metabolites in inflammation are:
i. Leukocytes.
ii. Mast cells.
iii. Endothelial cells.
iv. Platelets.
• These AA-derived mediators act locally at the site
of generation and then decay spontaneously or are
enzymatically destroyed.
 Prostaglandins and thromboxanes
1. Prostaglandin E2 (PGE2).
2. Prostaglandin D2 (PGD2).
3. Prostaglandin F2α (PGF2α).
4. Prostaglandin I2: PGI2 (Prostacyclin).
5. TXA2 (Thromboxane A2).
• Each derived by the action of a specific enzyme on
an intermediate.
 Prostaglandins and thromboxanes:
Actions
• PGD2 (along with PGE2 and PGF2α).
– Causes vasodilation.
– Potentiates edema formation by increase vascular
permeability.
• PGE2 also contribute to the pain and fever that
accompany inflammation.
• PGI2 (Prostacyclin)
– Vasodilator.
– Potent inhibitor of platelet aggregation.
• TXA2 (Thromboxanes)
– Potent platelet-aggregating agent.
– Vasoconstrictor. 22
 Leukotrienes (LT)
• Leukotriene B4 (LTB4)
– Produced by neutrophils and some macrophages.
– Potent chemotactic agent for neutrophils.
– Cause leukocyte activation and adhesion.
• Leukotriene C4, D4 and E4 (LTC4, LTD4 and
LTE4)
– Produced mainly in mast cells.
– They cause:
• Bronchoconstriction.
• Increased vascular permeability.
 Lipoxins
• Platelets that are activated and adherent to
leukocytes also are important sources of lipoxins.
• Lipoxins inhibit neutrophil chemotaxis and
adhesion to endothelium.
– Antagonists of leukotrienes B4 (LTB4).
 Platelet-activating factor
• Produced from leukocytes, mast cells.
• Causes:
i. Vasodilation.
ii. Increased vascular permeability.
iii. Leukocyte adhesion.
iv. Chemotaxis.
v. Degranulation.
vi. Oxidative burst.
 Cytokines
• Cytokines are polypeptide products of many cell
types that function as mediators of inflammation
and immune responses.
• The major cytokines in acute inflammation are:
i. Tumor Necrosis Factor (TNF).
ii. Interleukin-1 (IL-1).
iii. Interleukin-6 (1L-6).
iv. Chemokines (a group of chemoattractant
cytokines).
 Cytokines cont…
• Other cytokines that are more important in chronic
inflammation include:
i. Interferon-γ (IFN-γ)
ii. Interleukin-12 (IL-12).
• A cytokine called Interleukin-17 (IL-17),
produced by T lymphocytes and other cells, plays
an important role in recruiting neutrophils.
– It is involved in host defense against infections and in
inflammatory diseases.
 Tumor Necrosis Factor and
Interleukin-1
• Produced by activated macrophages, as well as
mast cells, endothelial cells.
• Their secretion is stimulated by microbial
products and products of T lymphocytes.
• The principal role of these cytokines in
inflammation is endothelial activation.
• Also stimulate the expression of adhesion
molecules on endothelial cells.
 Tumor Necrosis Factor and
Interleukin-1 cont…
• They can also induce the systemic acute-phase
reactions like fever, lethargy.
• TNF also increases the thrombogenicity of
endothelium.
• IL-1 activates tissue fibroblasts, resulting in
increased proliferation and production of
Extracellular Matrix (ECM).
 Chemokines
• The chemokines are structurally related proteins
that act primarily as chemoattractants for
different subsets of leukocytes.
• Their main functions of chemokines is recruit
leukocytes to the site of inflammation.
• They also activate leukocytes.
• Two (2) major groups are present:
– CXC chemokines.
– CC chemokines.
 Chemokines cont…

i. CXC chemokines
• E.g. Interleukin-8 (IL- 8).
• Produced by activated macrophages, endothelial
cells, mast cells, and fibroblasts.
• Causes:
– Chemotaxis.
– Phagocytosis.
ii. CC chemokines.
• E.g. Eotaxin
• Causes chemotaxis for eosinophils.
 Reactive Oxygen species (ROS)
• ROS are synthesized via the NADPH oxidase
(phagocyte oxidase) pathway.
– Released from neutrophils and macrophages that are
activated by microbes, immune complexes, cytokines, and
a variety of other inflammatory stimuli.
• Destroy phagocytosed microbes and necrotic cells.
– ROS are produced within lysosomes.
• When secreted at low levels, ROS can increase
chemokine, cytokine, and adhesion molecule
expression.
– Amplifying the cascade of inflammatory mediators.
32
 Reactive Oxygen species (ROS)
cont…
• At higher levels, these mediators are responsible for
tissue injury by several mechanisms, including:
1. Endothelial damage, with thrombosis and increased
permeability;
2. Protease activation and antiprotease inactivation, with
a net increase in breakdown of the Extracellular Matrix
(ECM); and
3. Direct injury to other cell types (e.g., tumor cells, red
cells, parenchymal cells).

33
 Reactive Oxygen species (ROS)
cont…
• Fortunately, various antioxidant protective
mechanisms (e.g., mediated by catalase, superoxide
dismutase, and glutathione) present in tissues and
blood minimize the toxicity of the oxygen
metabolites.
• Also Vitamins A, C, E are antioxidants.

34
 Nitric oxide (NO)
• An important function is a microbicidal
(cytotoxic) agent.
• Produced by activated macrophages.
• Other roles in inflammation, including:
i. Vasodilation.
ii. Antagonism of all stages of platelet activation
(adhesion, aggregation, and degranulation).
iii. Reduction of leukocyte recruitment at
inflammatory sites.
Lysosomal enzymes of leukocytes
• The lysosomal granules of neutrophils and
monocytes contain many enzymes that
– Destroy phagocytosed substances .
– Capable of causing tissue damage.
• Acid proteases generally are active only in the low-
pH environment of phagolysosomes.
• Neutral proteases including elastase, collagenase,
and cathepsin:
– Degrade extracellular matrix components.
– Cleave the complement proteins.
 Neuropeptides
• These are small proteins such as substance P.
• They:
– Transmit pain signals.
– Regulate vessel tone.
– Modulate vascular permeability.
• Nerve fibers that secrete neuropeptides are
especially prominent in the lung and GIT.
Plasma protein–derived mediators

• Circulating proteins of three interrelated systems.

i. Complements.
ii. Kinins.
iii. Coagulation proteins.
• They are all produced primarily in the liver.
A. Complement plasma causes:
i. Leukocyte chemotaxis and activation.
ii. Direct target killing.
iii. Vasodilation (mast cell stimulation).
Plasma protein–derived mediators cont…

B. Kinins plasma:
– Bradykinin is one of the example.
– They causes:
i. Increased vascular permeability.
ii. Smooth muscle contraction (esp. bronchial).
iii. Vasodilation.
iv. Pain.
C. Coagulation proteins causes:
i. Endothelial activation.
ii. Leukocyte recruitment.
 Anti inflammatory mechanisms
• Activated macrophages and other cells secrete a
cytokine, IL-10.
– IL-10 down-regulate the responses of activated
macrophages, thus providing a negative feedback loop.
• Other anti inflammatory cytokines include TGF-β.
– Also a mediator of fibrosis in tissue repair after
inflammation.
• Cells also express a number of intracellular proteins,
such as tyrosine phosphatases.
– Inhibit pro-inflammatory signals triggered by receptors that
recognize microbes and cytokines
40
 Anti inflammatory mechanisms
cont…
• Inflammatory reactions subside because many of the
mediators are short-lived and are destroyed by
degradative enzymes.
• In addition, there are several mechanisms that
counteract inflammatory mediators and function to
limit or terminate the inflammatory response.
– Some of these mediators are lipoxins and complement
regulatory proteins.

41
 Reactions of Inflammation and their
responsible Chemical Mediators
1. Vasodilation.
– Histamine.
– Prostaglandins (PGDE2, PGE2, PGI2).
– Nitric oxide.
– Substance P.

42
 Reactions of Inflammation and their
responsible Chemical Mediators
2. Increased vascular permeability (Vascular
leakage).
– Histamine.
– Serotonin.
– Bradykinin.
– Substance P.
– Leukotrienes C4, D4, E4.
– Platelet Activating Factor.
– C3a and C5a.
• NOTE:
– Prostaglandins potentiates other chemical mediators (like
Histamine) to cause vascular leakage. 43
 Reactions of Inflammation and their
responsible Chemical Mediators cont…
3. Chemotaxis, leukocyte recruitment and
activation
– Tumor Necrosis Factor.
– Interleukin-1.
– Chemokines (IL 8, Eotaxin).
– C3a, C5a.
– Leukotriene B4.
– Bacterial products (e.g., N-formyl methyl peptides).

44
 Reactions of Inflammation and their
responsible Chemical Mediators cont…
4. Fever.
– Tumor Necrosis Factor.
– Interleukin-1.
– Interleukin-6.
– Prostaglandins (especially PGE2).

45
 Reactions of Inflammation and their
responsible Chemical Mediators cont…
5. Pain.
– Prostaglandins.
– Bradykinin.

6. Tissue damage.
– Lysosomal enzymes of leukocytes.
– Reactive oxygen species.
– Nitric oxide.

46
 Key points
• Chemical mediators of inflammation can be derived
from cells or plasma.
• Prostaglandins and bradykinins induce pain.
• Fever is induced by TNF, IL-1, IL-6 and PGDE2.
• Prostaglandins potentiate other chemical mediators
(like Histamine) to cause vascular leakage.
• Most of chemical mediators have the potential to
cause harmful effects if are unchecked.

47
 Review questions
1. Mention three (3) general properties of chemical
mediators of inflammation.
2. List five (5) chemical mediators which cause
vascular leakage.
3. Mention three (3) chemical mediators which causes
tissue damage.
4. List three (3) chemical mediators which cause
vasodilation.
5. Enumerate three (3) reactions of inflammation
caused by histamine.
48
 References
• Bezabeh M.; Tesfaye A.; Ergicho B. et al (2004):
General pathology lecture notes for Health Sciences
students. Ethiopia Public Health Training Initiative.
Pg. 30-31.
• Kumar V.; Abbas A. K.; Aster J. C.;(2013): Robbins
and Contran Pathologic Basis of Disease (9th Ed.)
Elsevier Saunders, China. Pg. 44-52.
• Mohan H.;(2010): Text book of Pathology (6 th Ed.)
Jaypee Brothers Medical Publishers, India . Pg. 136-
140.
49