By Duy Thai, 1997 Pharmacology Semester 1 page 1 of 4

ENDOTHELIUM DERIVED VASOACTIVE FACTORS

Functions of the endothelium

1. A barrier separating the vascular smooth muscle from the luminal contents.
• By acting as a barrier, it also prevents adhesion of blood cells to the lining of the blood vessel (via release
of various factors)
2. Metabolic role
• The endothelium has a number of enzymes. e.g. Serotonin released from the platelets is metabolised by
the endothelium.
3. Source of vasoactive chemicals
A. PGI2
• Predominantly synthesised in the endothelium
• A vasodilator
• Antiaggregatory to platelets
B. Endothelium derived relaxing factor (NO)
C. Endothelin

Endothelium derived relaxing factor

• When Ach is given intravenously in vivo, it causes vasodilation
• When Ach is present in an organ bath in vitro, it causes vasoconstriction

• To explain this is as follows:

• If the endothelium of the blood vessel is intact, Ach acts on receptors on the endothelial cells to produce
NO. It is the NO which diffuses to the underlying smooth muscle to cause vasodilation
• If the endothelium is absent, Ach acts directly on the smooth muscle to cause vasoconstriciton.

• At very high concentrations of Ach, it starts to contract.

• Normally, Ach can cause vasodilation (via NO) and vasoconstriction (direct action) simultaneously. At
low concentrations, the balance favours vasodilation over vasoconstriction. However, at high
concentrations, the balance is reversed, so that there is more vasoconstriction than dilation.

Other vasodilator substances

Endothelium dependent Endothelium independent

Bradykinin Nitrovasodilators (GTN)
Substance P Isoprenaline (beta agonist)
Histamine PGI2
Serotonin

• In addition to mediating vasodilatin on its own, NO can also act as a modulator to vasoconstriciton.

• e.g. If NA is present, it will cause a vasoconstriction. However, if NO is present, the vasoconstriction will be less
strong. If the endothelium were damaged, there will be no NO to modulate the vasoconstriction of NA, and so you
get a higher level of contraction (hypertension)

Functions of NO/EDRF
• Relaxes blood vessels
• Inhibits platelet aggregation and adhesion
• Inhibited by hemoglobin
• NO is rapidly mopped up by RBC, thus preventing it from circulating and also causing it to be very shory
acting
• Reacts with oxygen, making it less able to circulate
• Oxygen radical scavengers can potentiate its action
• Stimulates guanylate cyclase
• Has a very short half life (4 sec)
By Duy Thai, 1997 Pharmacology Semester 1 page 2 of 4

Synthesis of NO

• L arginine + O2 ----NOS-----> NO + L citrulline + H+

• Since L arginine is available everywhere, the rate limiting step is the activity of NOS (NOS determines whether
NO is produced or not)

• There are 2 forms of NOS:

1. A constitutive form
2. An inducible form

• Comparison of the 2 forms:

Constitutive form Inducible form

Constant activity Only active when stimulated
Found in: Found in:
Endothelium Macrophages
Nerves Vascular smooth muscle
Produces low [NO] Produces high [NO]
Short lasting Sustained

• NO can also be cytotoxic, and hence contribute to inflammatory conditions.

How to inhibit endothelium dependent dilation

1. Remove the endothelium
2. Bind NO (by hemoglobin)
3. Inhibit guanylate cyclase
• Not practical becaue guanylate cyclase is involved in many other functions
4. Inhibit NOS
• Via analogues of L arginine

Effects of NOS inhibition

• Vasoconstriction
• Hypertension

• The basal activity of NO is to modulate the vascular tone (preventing excessive systemic vasoconstriction). Thus,
NO is always being released.
• If NO is stopped being released (by some pathological reason), then hypertension may result.

Physiological role of NO
1. Flow dependent vasodilation
• If there is increased blood flow over the surface of an artery, its diameter will increase due to a more rapid
flow.
• The increase in shear forces will stimulate the release of NO. Thus, you have autoregulation of flow.

2. Inhibits platelet adhesion and aggregation

• By having more rapid flow and turbulence, you increase the likelihood of platelets sticking to the surface.
The NO released will prevent this from happening, thus preventing thrombus formation
3. Neurotranmitter
• In CNS and periphery

Endothelial dysfunction (pathological inactivation)

• Atherosclerosis, hypertension and diabetes can all lead to impairment of endothelial dependent vasodilation.
• However, the question is: is endothelial dysfunction because of these diseases, or does it lead to the diseases. We
don’t know yet!
By Duy Thai, 1997 Pharmacology Semester 1 page 3 of 4

Impaired response to Ach

• A study was done to look at the effects of Ach in coronary arteries. These arteries had no evidence of
atherosclerosis. These people all had the risk factors accociated with atherosclerosis:
• Old age, elevated cholesterol, smoking and hypertension
• What they found was a negative correlation between the amount of risk factors vs. the effects of Ach on the
coronary blood vessel. Recall that these blood vessels seemed normal in that they did not show atheroma.

Effects of Ach on the coronary vessels

Dilation (intact
endo)

Constrict
(dysfunctional endo)
No. Of risk factors
• What this shows is that exposure to multiple risk factors will lead to progressive endothelial dysfunction,
possibly resulting in coronary artery disease. Ach lost the ability to dilate arteries to increased flow,
therefore you get inappropriate vasoconstriction. Also, an absence of NO will increase the likelihood of
thrombus formation.

Endotoxic shock
• Resistance to vasoconstriction
• Due to stimulation of iNOS (inducible form of NOS) via inflammatory cells
• Vasodilation, reduced sensitivity to vasoconstrictor agents (vasoconstrictors don’t work)
• However, the hypotension is reversed by NOS inhibitors
• This results in restoration of blood pressure.
• However, the NOS inhibitors are not selective for iNOS or cNOS, and so you also get inhibition of
the constitutive form. This can result in constriction of renal arteries, leading to renal failure

Therapeutic uses
• Pulmonary hypertension
• Inhale NO gas to cause vasodilation of the pulmonary vessels
• Won’t have a systemic action because of the shory half life (hence you won’t get systemic
hypertension)
• Angina
• Use drugs which donate NO (glycerol trinitrate)
• NO is released directly from the drug, hence you do not need the endothelium to be intact to get
vasodilation

ENDOTHELIN

• Endothelin is the most potent vasoconstrictor agent

• Comes in a variety of isoforms
• ET1, ET2, ET3
• ET1 is mainly synthesised by endothelial cells

Synthesis
• Big ET1 ET1
• The enzyme is endothelin converting enzyme
• This enzyme is the target of drugs

Endothelin receptors
• ETA ET1=ET2 > ET3
• ETB ET1=ET2=ET3

• ETA and ETB mediate vasoconstriction

• ETB mediates NO and PGI2 (hence vasodilation)

• Endothelin cuases a profound vasoconstriction but is modulated by NO and PGI2 via ETB receptor
By Duy Thai, 1997 Pharmacology Semester 1 page 4 of 4

• Endothelin is also a potent bronchoconstrictor

Physiological role of endothelin

• May be involved in:
• Ischaemia
• Congestive heart failure
• Pulmonary hypertension
• In all these conditions, you can see a rise in circulating endothelin (since it is not broken down quickly, it
can circulate - it is a stable peptide). NO on the other hand cannot circulate because it only has a half life of
4 sec
• Antagonists of endothelin receptors:
• May be useful as an antihypertensive and also decrease the vasoconstriction in ischaemia.
• Also a treatment of asthma?