Chemical​ ​Mediators​ ​of​ ​Inflammation

Chemical​ ​Mediators​ ​are​ ​the​ ​substances​ ​that​ ​initiate​ ​and​ ​regulate​ ​(control)
inflammatory​ ​reactions.​ ​(like​ ​an​ ​orchestra).

Chemical​ ​mediators​ ​are​ ​subdivided​ ​according​ ​to​ ​their​ ​origin​ ​(source),​ ​we
divide​ ​them​ ​just​ ​to​ ​simplify.

They​ ​are​ ​subdivided​ ​into:

● Plasma​ ​derived​:​ ​like​ ​plasma​ ​proteins​,​ ​which​ ​are​ ​synthesized​ ​in
the​ ​liver,​ ​present​ ​in​ ​an​ ​inactive​​ ​form​ ​and​ ​normally​ ​circulate​ ​(in​ ​the
plasma)​ ​as​ ​inactive​ ​precursors​ ​(​pro-proteins​)​.​ ​Once​ ​inflammation
occurs​ ​they​ ​will​ ​be​ ​activated.
​ ​Such​ ​as:​ ​Coagulation/fibrinolytic​ ​factors,​ ​complement​ ​system​ ​and​ ​kinin​ ​system

● Cell​ ​derived:​ ​they​ ​come​ ​(derived)​ ​from​ ​cells,​ ​most​ ​come​ ​from
leukocytes​.​ ​Cell-derived​ ​chemical​ ​mediators​ ​are​ ​subdivided​ ​into
two​ ​types:
o Type​ ​which​ ​is​ ​usually​ ​there​​ ​inside​ ​the​ ​cell​ ​-​ ​either​ ​if​ ​there
is​ ​an​ ​inflammation​ ​or​ ​not​ ​–​ ​and​ ​they​ ​are​ ​sequestered​ ​in
granules.​ ​Like​ ​Vasoactive​ ​amines​ ​(serotonin​ ​and​ ​histamine)

o Type​ ​which​ ​is​ ​not​ ​usually​ ​there​,​ ​synthesized​ ​upon​ ​to

stimulation.​ ​Like​ ​PGs,​ ​LT,​ ​O​2​​ ​species,​ ​NO,​ ​Cytokines,​ ​and
PAF

The​ ​Figure​ ​in​ ​next​ ​page​ ​is​ ​very​ ​important​ ​to

understand​​ ​the​ ​types​ ​of​ ​chemical​ ​mediators,
and​ ​you​ ​should​ ​know​ ​the​ ​source​ ​of​ ​each​ ​CM,
the​ ​Doctor​ ​read​ ​them​ ​with​ ​no​ ​additions.
 General​ ​characteristics​ ​of​ ​CM:
● They​ ​act​ ​by​ ​binding​ ​to​ ​specific​ ​cellular​ ​receptors,​ ​or​ ​have​ ​enzymatic
activity.
● This​ ​binding​ ​may​ ​stimulate​ ​target​ ​cells​ ​to​ ​release​ ​secondary
mediators​ ​with​ ​similar​ ​or​ ​opposing​ ​functions​.​ ​The​ ​secondary
mediator​ ​with​ ​similar​ ​activity​ ​of​ ​the​ ​first​ ​one​ ​will​ ​propagate​ ​the
process,​ ​after​ ​a​ ​while​ ​the​ ​first​ ​mediator​ ​will​ ​stimulate​ ​the​ ​target​ ​cell
to​ ​release​ ​a​ ​mediator​ ​with​ ​an​ ​opposite​ ​function​ ​to​ ​inhibit​ ​the
process.​ ​Just​ ​like​ ​negative​ ​feedback​ ​mechanism​.
● Some​ ​have​ ​limited​ ​targets​ ​and​ ​others​ ​have​ ​wide​ ​spread​ ​activities.
● Each​ ​chemical​ ​mediator​ ​has​ ​an​ ​inhibitory​ ​mechanism​.​ ​It’s​ ​important
for​ ​each​ ​mediator​ ​to​ ​have​ ​an​ ​inhibitor,​ ​otherwise​ ​the​ ​inflammation
will​ ​be​ ​unchecked,​ ​and​ ​would​ ​have​ ​a​ ​harmful​ ​effect​​ ​(it​ ​will​ ​destroy
and​ ​act​ ​on​ ​normal​ ​tissue).

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​ ​Examples​ ​of​ ​inhibitory​ ​mechanisms:
▪ Short​ ​half-life​ ​(AA​ ​(Arachidonic​ ​Acid)​ ​metabolites)
▪ Inactivated​ ​by​ ​enzymes​ ​(kininase​ ​on​ ​bradykinin)
▪ Eliminated​ ​(antioxidants​ ​on​ ​O2​ ​species)
▪ Inhibitory​ ​proteins​ ​(just​ ​like​ ​in​ ​the​ ​complementary
system,​ ​specific​ ​to​ ​inhibit​ ​complement​ ​protein)

Now​ ​we​ ​will​ ​discuss​ ​Cell-derived​ ​chemical​ ​mediators​ ​(Histamine​ ​and

Serotonin).

Vasoactive​ ​Amines​ ​(1​st​​ ​Type​ ​of​ ​Cell-Derived​ ​CM)

There​ ​are​ ​many​ ​factors​ ​that​ ​will​ ​lead​ ​to​ ​the​ ​release​ ​of​ ​vasoactive​ ​amines
such​ ​as​ ​histamine​ ​and​ ​serotonin​ ​in​ ​response​ ​to​ ​the​ ​inflammatory​ ​process.

For​ ​Histamine:​ ​(these​ ​points​ ​would​ ​induce​ ​mast​ ​cells​ ​to​ ​secrete​ ​histamine)
● Physical​ ​injury.
● Binding​ ​of​ ​IgE​ ​to​ ​the​ ​FC​ ​receptors.
● The​ ​release​ ​of​ ​anaphylatoxins​ ​(C3a,​ ​C5a)​ ​from​ ​the​ ​complement
system.
● Histamine​ ​releasing​ ​protein​ ​which​ ​is​ ​derived​ ​from​ ​PMNs
(polymorphonuclear​ ​leukocytes).
● Neuropeptide.
● Cytokines;​ ​interleukin-1​ ​(IL-1)​ ​&​ ​interleukin-8​ ​(IL-8).

For​ ​Serotonin:
● Platelets​ ​aggregation
● PAF​ ​(​Platelet​ ​activating​ ​factor​)

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 ​ ​ ​ ​ ​What​ ​are​ ​the​ ​functions(effects)​ ​of​ ​Serotonin​ ​&​ ​Histamine:

Histamine​ ​which​ ​is​ ​found​ ​inside​ ​the​ ​granules​ ​in​ ​mast​ ​cells​ ​and​ ​serotonin
inside​ ​the​ ​platelets​ ​cause​ ​arteriolar​ ​dilatation​​ ​and​ ​increase​ ​of​​ ​vascular
permeability​*​ ​(​immediate​ ​phase​ ​reaction​).​ ​They​ ​also​ ​induce​ ​endothelial​ ​cell
contraction​ ​in​ ​venules​​ ​by​ ​binding​ ​to​ ​H1​ ​receptors,​ ​Histamine​ ​is​ ​inactivated​ ​by
histaminase​.

*​Achieved​ ​by​ ​endothelial​ ​cell​ ​contraction(immediate​ ​transient​ ​response;​ ​begins

immediately​ ​and​ ​ends​ ​quickly)

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 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​Arachidonic​ ​acid​ ​(2​nd​​ ​Type​ ​of​ ​Cell-Derived​ ​CM)

Arachidonic​ ​acid​ ​is​ ​a​ ​ ​component​ ​of​ ​the​ ​cell​ ​membrane;​ ​it​ ​needs​ ​to​ ​be
released​ ​from​ ​the​ ​membrane​ ​to​ ​the​ ​cytoplasm​ ​by​ ​an​ ​enzyme​ ​called
phospholipase​ ​A2.​ ​So,​ ​activating​ ​phospholipase​ ​A2​ ​will​ ​work​ ​on​ ​the​ ​phospholipids
of​ ​the​ ​membrane​ ​to​ ​ ​release​ ​arachidonic​ ​acid​ ​metabolites​ ​production​,​ ​and​ ​also
will​ ​increase​ ​the​ ​production​ ​of​ ​platelet​ ​activating​ ​factor(PAF).

Now​ ​we​ ​have​ ​the​ ​arachidonic​ ​acid​ ​in​ ​the​ ​cytoplasm,​ ​there​ ​are​ ​two​ ​families
of​ ​enzymes​ ​that​ ​can​ ​act​ ​on​ ​it:
● Cyclooxygenases​ ​pathway.
● Lipoxygenase​ ​pathway.
Cyclooxygenase​ ​Pathway:

● Produces​ ​prostaglandins​ ​“PG”.

● These​ ​prostaglandins​ ​are:​ ​PGE2,​ ​PGD2,​ ​PG-F2a,​ ​PGI2,​ ​Thromboxane​ ​A2
(TX-A2).
PG-E2​ ​and​ ​PG-D2​ ​and​ ​PG-F2a:​ ​-​Vasodilators​,​ ​increase​ ​permeability,​ ​they​ ​cause
pain,​ ​fever​ ​and​ ​edema.​ ​(Fever​ ​because​ ​they​ ​interact​ ​with​ ​cytokines).

PG-I2:​ ​-​ ​Vasodilator​,​ ​inhibits​ ​platelet​ ​aggregation​ ​(which​ ​causes​ ​bleeding).

● TXA2:​ ​-Secreted​ ​by​ ​platelets,​ ​vasoconstrictor​,​ ​increases​ ​platelet

aggregation​ ​(blood​ ​clotting).
● Note​ ​that​ ​although​ ​PG12​ ​and​ ​TXA2​ ​come​ ​from​ ​the​ ​same​ ​source​ ​they​ ​have
opposing​ ​effects.
● The​ ​balance​ ​between​ ​PGI2​ ​AND​ ​TXA2​ ​is​ ​very​ ​important.
● If​ ​PGI2​ ​increased,​ ​it​ ​will​ ​cause​ ​bleeding.​ ​And​ ​if​ ​TXA2​ ​increased,​ ​it​ ​will​ ​cause
over​ ​clotting.

Lipoxygenase​ ​pathway
We​ ​have​ ​two​ ​enzymes:
1) 5-lipoxygenase​ ​gives​ ​us:
● 5-HETE​ ​Leukotrienes,​ ​they​ ​are:​ ​LT-A4​ ​/​ ​LT-B4​ ​/​ ​LT-C4​ ​/​ ​LT-D4​ ​/​ ​LT-E4
● LT-A4​ ​produces​ ​LT-​ ​B4
● 5HETE​ ​&​ ​LTB4​ ​are​ ​the​ ​only​ ​chemotactic​ ​agents.

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 ● LT-(C4,​ ​D4,​ ​E4):​ ​Broncho-constrictors​​ ​(Bronchospasm),​ ​increase​ ​vascular
permeability​ ​and​ ​vasoconstriction.
2)​ ​ ​12-lipoxygenase​ ​gives​ ​us:
● Lipoxins:​ ​LX-A4,​ ​LXB4.
- Anti-inflammatory​ ​agents​ ​(Reduce​ ​inflammation)​ ​by​ ​point2.
- Inhibit​ ​neutrophil​ ​adhesion​ ​and​ ​chemotaxis.
- Stimulation​ ​of​ ​monocyte​ ​adhesion​ ​for​ ​repair

● Steroids:​ ​(as​ ​a​ ​drug​ ​to​ ​treat​ ​many​ ​inflammatory​ ​conditions)

Block​ ​the​ ​action​ ​of​ ​phospholipase​ ​A2,​ ​thus​ ​steroids​ ​inhibit​ ​(deactivate)​ ​both
cyclooxygenase​ ​and​ ​lipoxygenase​ ​pathways​ ​and​ ​hence​ ​all​ ​the​ ​products​ ​of​ ​these
pathways​ ​will​ ​not​ ​be​ ​produced.​ ​They​ ​also​ ​inhibit​ ​“Platelet​ ​Activating​ ​Factor”
formation.
EX:​ ​Glucocorticoids​.

● Non-Steroidal​ ​anti-inflammatory​ ​drugs:

Inhibit​ ​COX​ ​(Cyclooxygenase)​ ​enzyme​ ​(COX-1​ ​&​ ​COX-2),​ ​so​ ​that​ ​will​ ​inhibit​ ​PG
production,​ ​so​ ​no​ ​pain​ ​would​ ​be​ ​there​ ​and​ ​no​ ​fever.​ ​PG-2​ ​(Prostaglandin-2)​ ​are
responsible​ ​for​ ​the​ ​sense​ ​of​ ​pain
during​ ​inflammation​.
Examples:​ ​Aspirin,​ ​and​ ​Ibuprofen.

In​ ​cases​ ​of​ ​asthma,we​ ​use

anti-leukotriene​ ​drugs​ ​we​ ​should
inhibit​ ​LT-(C4,​ ​D4,​ ​E4)​ ​because​ ​they
induce​ ​bronchospasm,​ ​so​ ​we​ ​should
use​ ​lipoxygenase​ ​inhibitors.​ ​These
examples​ ​are​ ​the​ ​reason​ ​why​ ​we
study​ ​those​ ​pathways,​ ​to​ ​give​ ​the
best​ ​drug​ ​for​ ​treatment.
Asthma;​ ​certain​ ​allergens​ ​irritate
mast​ ​cells​ ​which​ ​produce​ ​mediators
to​ ​induce​ ​bronchospasm.

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 Platelet​ ​activating​ ​factor
• Generated​ ​from​ ​the​ ​membrane’s​ ​phospholipids​ ​by​ ​Phospholipase​ ​A2.
• Causes​ ​platelet​ ​aggregation​ ​and​ ​degranulation.
• Potent​ ​vasodilator​ ​and​ ​bronchoconstrictor​.
• Increases​ ​vascular​ ​permeability.

● Effects​ ​on​ ​leukocytes:

• Increase​ ​adhesion​ ​to​ ​endothelial​ ​cells.
• Has​ ​a​ ​chemotactic​ ​function.
• Degranulation​ ​(induce​ ​platelets​ ​to​ ​secrete​ ​serotonin).
• Oxygen​ ​bursts​ ​(induces​ ​production​ ​of​ ​oxygen​ ​reactive​ ​species).

Cytokines
● Hormone-like​ ​polypeptides​ ​produced​ ​by​ ​cells,​ ​involved​ ​in​ ​cell​ ​to​ ​cell
communication.
● Pleiotropic​ ​effects.
● Secretion​ ​is​ ​transient​(not​ ​continuous).
● Effects​ ​can​ ​be:​ ​Autocrine​:​ ​the​ ​cell​ ​itself​ ​will​ ​secrete​ ​the​ ​cytokines​ ​and​ ​the
cytokines​ ​will​ ​act​ ​on​ ​the​ ​same​ ​cell.​ ​Paracrine​:​ ​the​ ​cell​ ​itself​ ​will​ ​secrete​ ​the
cytokines​ ​and​ ​they​ ​will​ ​act​ ​on​ ​the​ ​neighboring​ ​cell.​ ​Endocrine​:​ ​the​ ​cell​ ​itself
will​ ​secrete​ ​the​ ​cytokines​ ​but​ ​the​ ​cytokines​ ​will​ ​be​ ​secreted​ ​into​ ​the
bloodstream​ ​to​ ​affect​ ​cells​ ​away​ ​from​ ​it.

Classes​ ​of​ ​cytokines

• Regulators​ ​of​ ​lymphocyte​ ​function

- IL-2​ ​stimulates​ ​proliferation
- TGF(beta)​ ​inhibits​ ​lymphocytes​ ​growth
• Primary​ ​responders​ ​to​ ​injury​ ​(innate​ ​immunity)
- IL-1​ ​&​ ​TNF
• Activators​ ​of​ ​cell​ ​mediated​ ​immunity
- INF-g​ ​&​ ​IL-12
• Chemotactic
- IL-8
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 • Hematopoietic​ ​growth​ ​factors​ ​(induce​ ​leukocyte​ ​and​ ​monocyte​ ​production
from​ ​bone​ ​marrow;​ ​as​ ​we​ ​are​ ​losing​ ​them​ ​in​ ​the​ ​inflammatory​ ​process​ ​and
they​ ​need​ ​to​ ​be​ ​replenished​)
- IL-3​ ​&​ ​GM-CSF​ ​(Gran​ulocyte-macrophage​ ​colony-stimulating​ ​factor​)

IL-1​ ​&​ ​TNF

• Produced​ ​mainly​ ​by​ ​macrophages
• Secretion​ ​stimulated​ ​by:​ ​bacterial​ ​products,​ ​immune​ ​complexes,
endotoxins,​ ​physical​ ​injury,​ ​other​ ​cytokines.​ ​(​each​ ​one​ ​will​ ​be​ ​in​ ​the​ ​area​ ​of
inflammation​ ​can​ ​induce​ ​macrophages​ ​to​ ​secrete​ ​IL-1​ ​&​ ​TNF)
• They​ ​affect​ ​everything(hence​ ​are​ ​very​ ​important),​ ​such​ ​as​ ​having​ ​effects​ ​on
endothelial​ ​cell​,​ l​ eukocytes​,​ f​ ibroblasts​,​ ​and​ ​are​ ​the​ ​cause​ ​of​ ​the​ ​acute
phase​ ​reactions​​ ​(feeling​ ​of​ ​being​ ​ill)​.

Extra
notes​ ​on​ ​the
figure​ ​above:
● IL-1​ ​&
TNF
induce
the​​ ​endothelial​ ​cell​​ ​itself​ ​to​ ​secrete​ ​IL-1,​ ​IL-8,​ ​IL-6​ ​and​ ​PDGF​ ​(​Platelet-derived
growth​ ​factor).
● Hemodynamic​ ​effect​ ​is​ ​a​ ​result​ ​of​ ​vasodilatation​ ​which​ ​if​ ​severe​ ​may​ ​lead
to
hypotension​ ​and​ ​shock;​ ​as​ ​there​ ​will​ ​be​ ​increase​ ​of​ ​leukocytes(neutrophils)
number​ ​in​ ​the​ ​periphery.

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● Especially​ ​in​ ​cases​ ​of​ ​bacterial​ ​infection​ ​if​ ​you​ ​carry​ ​out​ ​a​ ​CBC​ ​you​ ​will​ ​find
an​ ​increase​ ​in​ ​neutrophil​ ​numbers​ ​especially​ ​in​ ​peripheral​ ​blood.
● Release​ ​of​ ​acute​ ​phase​ ​proteins​ ​(c-reactive​ ​protein​ ​and​ ​fibrinogen),​ ​if
present​ ​in​ ​blood​ ​indicate​ ​the​ ​ongoing​ ​process​ ​of​ ​inflammation,​ ​secreted
through​ ​the​ ​release​ ​of​ ​IL-1​ ​and​ ​TNF.
● Fibroblast​ ​effects​ ​are​ ​very​ ​important​ ​in​ ​the​ ​process​ ​of​ ​healing​.​ ​Healing
could​ ​be​ ​a​ ​scar​ ​(collagen​ ​participates​ ​in​ ​scar​ ​formation).

Nitric​ ​Oxide

• Produced​ ​from​ ​arginine​​ b ​ y​ ​the​ ​effect​ ​of​ ​nitric​ ​oxide​ ​synthase​ ​(NOS)
• 3​ ​isoforms​ i​ n​ ​our​ ​body:​ ​nNOS,​ ​iNOS,​ ​eNOS
• Role​ ​of​ ​NO​ i​ n​ ​inflammation:​ ​(​anti-inflammatory​​ ​function)
• Vasodilator​ ​(smooth​ ​muscle​ ​relaxant).
• Antagonist​ ​of​ ​platelets​ ​adhesion,​ ​aggregation​ ​and
stimulation.
• Reduces​ ​leukocytes​ ​adhesion​ ​and​ ​recruitment.
• Has​ ​microbicidal​ ​activity​ ​in​ ​activated
macrophages.​ ​(it​ ​can​ ​kill​ ​microorganisms​ ​in
activated​ ​macrophages​ ​through​ ​generation​ ​of
reactive​ ​oxygen​ ​species).

Note:​ ​*inos​ ​is​ ​mainly​ ​found​ ​in

macrophages​ ​but​ ​also​ ​present
in​ ​leukocytes​ ​and​ ​other​ ​cells
*the​ ​Dr.​ ​said​ ​to​ ​make​ ​things
easier​ ​memorise​ ​the​ ​mediators
that​ ​cause​ ​vasoconstriction
and​ ​bronchospasm​ ​since​ ​they
are​ ​fewer​ ​and​ ​then​ ​the​ ​rest..

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