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General And Systematic Pharmacology 11_TDC_SmMusc[570]

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General And Systematic Pharmacology 11_TDC_SmMusc[570]
UNIVERSITY COLLEGE LONDON

Department of Pharmacology



PHAR2002/2005 General and Systematic Pharmacology Dr T. D. Carter



SMOOTH MUSCLE CONTRACTION & RELAXATION



Learning objectives



1. To understand different ways in which vascular, bronchial and uterine smooth

muscle can be contracted or relaxed.



2. To give examples (where possible) of drugs acting through the mechanisms detailed in

(1) above.



3. To understand the rationale behind the therapeutic application of drugs affecting

smooth muscle tone.



Learning task



1. Supplement the lecture by reading Rang, Dale & Ritter (3rd Ed.) pps. 203-213, 301-313,

358-363 and 469-473 or the more diffuse treatment of the material covered in these

lecture notes in Katzung (5th Ed.)



Vasoconstriction (and vasodilatation) evoked by catecholamines is dealt with in other Lectures

(Adrenoceptors) and (Antihypertensive drugs).



Smooth muscle contraction



The contractile characteristics and the mechanisms that cause contraction of vascular smooth

muscle (VSM) are very different from cardiac muscle. VSM undergoes slow, sustained, tonic

contractions, whereas cardiac muscle contractions are rapid and of relatively short duration (a

few hundred milliseconds). While VSM contains actin and myosin, it does not have the

regulatory protein troponin as is found in the heart. Furthermore, the arrangement of actin

and myosin in VSM is not organized into distinct bands as it is in cardiac muscle. This is not

to imply that the contractile proteins of VSM are disorganized and not well-developed. They

are actually highly organized and well-suited for their role in maintaining tonic contractions

and reducing lumen diameter.





Mechanism



An increase in free intracellular calcium is the trigger for contraction. The free calcium binds

to a special calcium binding protein called calmodulin (see Fig.below). Calcium-calmodulin

activates myosin light chain kinase (MLCK), an enzyme that is capable of phosphorylating

myosin light chains (MLC) in the presence of ATP. Myosin light chains are 20-kD regulatory

subunits found on the myosin heads. MLC phosphorylation leads to cross-bridge formation

between the myosin heads and the actin filaments, and hence, smooth muscle contraction.





16-Oct-07/TDC 1

The intracellular concentration of

calcium depends upon the balance

between the calcium that enters the cell

from the external environment, the

calcium that is released by intracellular

storage sites (e.g., SR), and the removal

of calcium either back into storage sites

or out of the cell. Calcium is re-

sequestered by the SR by a ATP-

dependent calcium pump. Calcium is

removed from the cell to the external

environment by either a ATP-dependent

calcium pump or by the sodium-calcium exchanger.





Intracellular calcium concentration is very important in regulating smooth

muscle contraction.





Vascular smooth muscle (VSM) contraction





Blood Vessels;



Arteries are large diameter, thick-walled vessels that carry blood away from the heart.

Arterioles are small, thick-walled vessels that represent the major part of vascular resistance.

These resistance vessels serve as "circulatory stopcocks"and control the distribution of blood

to various organs.

Capillaries are extremely small, extremely thin-walled vessels (one cell thick) that allow

exchange of gases, nutrients, and other small molecules between the blood stream and

tissues. Increases in capillary hydrostatic pressure or capillary permeability can lead to

edema.

Venules are small thin-walled vessels that serve to bring blood back to the heart. These

vessels are highly distensible and (along with veins) contain a large fraction of the blood

volume.

Veins are large diameter thin-walled vessels that bring blood back to heart. They are

distensible and (in addition to venules) contain a large fraction of the blood volume.



The important vessels controlling blood pressure



Resistance vessels. Arterioles are the primary resistance vessels and control mean arterial

blood pressure and blood flow to specific tissues. Vascular smooth muscle tone in these

vessels is controlled by the sympathetic nervous system and local factors (metabolic need)

Capacitance vessels. Systemic venules and veins serve as a volume reservoir for the

circulatory system (approx. 50% of total blood volume is contained in these vessels).

Sympathetic and humoral regulation of these vessels can significantly alter venous return

(preload) and fluid exchange in the associated capillary beds.







16-Oct-07/TDC 2

Contraction in VSM can be initiated by mechanical, electrical, and chemical stimuli. Passive

stretching of VSM can cause contraction that originates from the smooth muscle itself and is

therefore termed a myogenic response. Electrical depolarization of the VSM cell membrane

will also elicit contraction, most likely by opening voltage dependent calcium channels (L-

type calcium channels) and causing an increase in the intracellular concentration of calcium.

Finally, a number of chemical stimuli such can elicit contraction;



noradrenaline (α1-adrenoceptor)

angiotensin II

endothelin (ETA, ETB2)

vasopressin (V1 receptor)

ergot alkaloids e.g. ergotamine

5-hydroxytryptamine (5-HT1D-like receptor)



Each of these substances bind to specific receptors on the VSM cell (or to receptors on the

endothelium adjacent to the VSM) and can cause contraction of the VSM. The mechanism of

contraction can involve different signal transduction pathways (see below) all of which

converge to increase intracellular calcium (see below).



Endothelium

Noradrenaline ATP

Endothelin Na

Angiotensin II

G-protein-coupled L-type Ca Ca

Non-selective cation

receptor channel channel

Gq +

GDP Depolorisation

PLC GTP





InsP3 [Ca2+]i

PL

+ CaM Ca-CaM

+

Calcium MLCK



Store (SR)



Contraction



Smooth muscle cell

16-Oct-07/TDC 3

Noradrenaline (NA)



From sympathetic nerves acting via alpha1-adrenoceptors coupled via Gq to PLC and InsP3

production. Co-transmitters may include; ATP that can cause contraction acting via

activation of a non-selective cation channel (P2x.receptors- very fast) or Gq coupled

receptors to PLC (e.g. P2y receptors—slow), Neuropeptide Y, mechanism of action not clear

but can potentiate action of NA.

alpha1-adrenoceptor antagonists acts as vasodilators, e.g. Prazosin, Indoramine. Cause

vasodilation and a fall in blood pressure.

Indirect acting vasoconstrictors that cause NA release from nerve terminals include

amphetamine, tyramine (e.g ‘the cheese reaction’) and ephedrine. Cocaine blocks uptake of

NA into nerve terminals and has a similar sympathomimetic effect.





Angiotensin II (AII).



A potent vasoconstrictor formed from the inactive angiotensin I by the angiotensin converting

enzyme (ACE; expressed onvascular endothelial cells). This enzyme is inhibited by captopril

which is an anti-hypertensive drug. AII, acts via ATI receptors coupled via Gq to PLC and

InsP3 production.



Endothelin (ETA, ETB2)



21 aa peptide, 3 isoforms ET1, ET2 and ET3. ET1 made by endothelium. ET1 acts via ETA and

ETB2 receptors that are coupled by Gq to PLC and InsP3 production. Antagonists include BQ-

123 (a cylic pentapeptide) and BMS 182874 (a sulphonamide derivative).



Vasopressin (VP).



VP is a posterior pituitary peptide hormone with major actions on the kidney (see Chp 20 Rang,

Dale & Ritter) It vascular actions are via V1 receptors to elicit VSM contraction (via Gq, PLC

and InsP3 production). Will also constrict gastrointestinal and uterine SM). The vasopressin

analog, felypressin (selective for V1 receptors) is used as a vasoconstrictor with local

anaesthetics.



Ergot alkaloids, 5-hydroxytryptamine and Migraine



Migraine is a common condition characterized by a headache and contraction and/or dilatation of

cerebral blood vessels. The symptoms have been treated with ergotamine which causes marked

vasoconstriction (must be avoided in patients with peripheral vascular disease). Very recently

the 5-HT1D-like receptor agonist sumatriptan, which also constricts intracranial vascular smooth

muscle, has been introduced. It has been suggested that migraine is a neurogenic inflammatory

condition mediated by nerves from the trigeminal nucleus. Sumatriptan may act prejunctionally

on these nerves which innervate cerebral blood vessels. The complex role of 5-HT in migraine is

shown by the prophylactic use of methysergide, which has some selectivity as an antagonist for

5-HT2 receptors.





---------------------------------------------------------------------------------------------------------------------



16-Oct-07/TDC 4

VSM relaxation:



The G-protein coupled pathway can either stimulate (via Gs protein) or inhibit (via Gi

protein) adenylyl cyclase (AC) that catalyzes the formation of cAMP. In VSM, unlike the

heart, an increase in cAMP (e.g., a beta-agonist such as epinephrine or isoproterenol) causes

relaxation. The mechanism for this is cAMP inhibition of MLCK. This decreases MLC

phosphorylation, thereby decreasing the interactions between actin and myosin. Therefore,

drugs which increase cAMP (e.g., β2-adrenoceptor agonists, phosphodiesterase inhibitors)

cause vasodilation.









A third mechanism that is

very important in

regulating VSM tone is the

nitric oxide (NO)-cGMP

system. Briefly, increases

in NO activate guanylyl

cyclase causing increased

formation of cGMP and

vasodilation. The precise

mechanisms by which

cGMP relaxes VSM is

unclear; however, cGMP

can activate a cGMP-

dependent protein kinase,

inhibit calcium entry into the VSM, activate K+ channels, and decrease IP3.



VSM vasodilators include;



adrenaline (β2-adrenoceptor)

nitrovasodilators e.g. glyceryl trinitrate

nitric oxide

calcium antagonists e.g. nifedipine

potassiun channel openers

histamine (H1 and H2 receptors; Lecture 14)

prostacyclin (Lecture 16)



The nitrovasodilators relax vascular smooth muscle by releasing nitric oxide (NO). The NO

stimulates guanylate cyclase which leads to an increase in cyclic GMP in the smooth muscle.

Nitric oxide is a gas (not to be confused with nitrous oxide, N2O) and has recently been given to

relieve respiratory distress syndrome. Some drugs act via receptors on the endothelium to

stimulate NO synthase and the NO produced diffuses to the smooth muscle to evoke relaxation.

Inhibitors of NO synthase, which increase blood pressure, are being tested in septic shock.





16-Oct-07/TDC 5

Pelvic nerve stimulation leads to the release of nitric oxide and subsequent relaxation of the

smooth muscle of the corpus cavernosum.. The nitric oxide is metabolised by a

phosphodiesterase, PDE-5. Sildenafil (Viagra) inhibits this enzyme so prolonging the effect of

nitric oxide stimulated cGMP.



The calcium antagonists block voltage-gated calcium channels and prevent calcium entry. In

smooth muscle the L-type channels are responsible for the inward movement of calcium that

plays a role in contraction. The dihydropyridine derivative nifedipine evokes arteriolar dilatation

and a fall in blood pressure with a transient reflex tachycardia.



Smooth muscle relaxation can also occur by increasing the membrane permeability to potassium

ions, leading to hyperpolarization. Drugs with this type of action are being developed.





Bronchial smooth muscle



A. Contraction:

acetylcholine (M3 muscarinic receptor)

some neuropeptides e.g. neurokinin A

leukotrienes C4 and D4

histamine (H1-receptor)



The bronchi are contracted by acetylcholine released from the postganglionic parasympathetic

nerve terminals. The muscarinic receptor antagonist, ipratropium bromide, is used as an anti-

asthmatic drug and is given by aerosol inhalation. It is a quaternary compound and therefore,

because it has low lipid solubility, it is not well absorbed into the circulation and does not readily

cross the blood-brain barrier.



B. Relaxation:

salbutamol (β2-adrenoceptor)

methyl xanthines e.g. caffeine and theophylline





β2-Adrenoceptor agonists are widely used to dilate bronchial smooth muscle (acting as

"physiological" antagonists) and they also inhibit the release of mediators from mast cells. These

drugs are usually taken by inhalation and have a quicker onset of action than ipratropium.



The methyl xanthines are bronchodilators and are given orally. Theophylline is often used,

sometimes complexed with ethylene diamine and known as aminophylline. Other effects which

also contribute to the use of aminophylline in left ventricular failure with pulmonary oedema are

positive chronotropic and inotropic effects on the heart, vasodilatation and the weak diuretic

activity. The mechanism of action of the methyl xanthines is unclear. Inhibition of

phosphodiesterase and the release of intracellular calcium have been shown but at fairly high

concentrations. A more likely mechanism for therapeutic drug doses is the antagonism of

adenosine at adenosine receptors.



Uterine smooth muscle



A. Contraction:



16-Oct-07/TDC 6

noradrenaline (α1-adrenoceptor)

acetylcholine (muscarinic receptor)

oxytocin (pregnant uterus)

ergometrine

prostaglandin E2 and F2α



The posterior pituitary hormone oxytocin increases the force and rate of contractions of the

pregnant uterus, with increased receptor numbers (oestrogen dependent) towards term. Oxytocin

may have a role in the initiation of paturition. At term oxytocin is often given by a slow,

intravenous infusion to induce labour. It causes regular contractions but the uterus relaxes in

between, allowing oxygenated blood flow to the foetus. In higher doses oxytocin can be used to

reduce post-partum haemorrhage (see below).



The ergot alkaloid ergometrine evokes contraction of smooth muscle including the pregnant and

non-pregnant uterus. The sustained contraction is unsuitable to induce labour. It is given,

intramuscularly or intravenously, in the third stage of labour (often combined with oxytocin) to

contract the uterus and to reduce the risk of post-partum haemorrhage.



The prostaglandins E2 and F2α cause marked rhythmical contractions of the pregnant and non-

pregnant uterus. Given by the extra-amniotic route these drugs are used in the second trimester

of pregnancy for therapeutic abortion.



B. Relaxation:

salbutamol (β2-adrenoceptor)



In some cases of premature labour the β2-adrenoceptor agonists are used to relax uterine smooth

muscle.









16-Oct-07/TDC 7


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