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Course Medpharma lab - Medpharma laboratory
Lesson Airways Resistance – Lab
Student email kibtabungar2019@plm.edu.ph

Group Activity

Kaezzy Ila B. TABUNGAR GROUP LEADER

Lesson committed 6 Jan 2021  11:34 am

Lesson started 6 Jan 2021  8:40 am

Asthma – Challenge

The schematic below shows the ligands, receptors, and pathways that produce the symptoms of asthma: constriction of
bronchial smooth muscle and mucus secretion. Place the labels to indicate where the following drugs act to reduce the
symptoms of asthma.

Ipratroprium Salbutamol Theophylline

(an antagonist) (a selective agonist) (a methylxanthine)

Salbutamol
(a selective agonist)

Ipratroprium
(an antagonist)

Theophylline
(a methylxanthine)
Force transducer setup

Force

Time

Tissue tension setup

What volume (in mL) of a 6 × 10-5 M stock solution would be required to give a nal concentration of 3 × 10-6 M in a 30
mL organ bath?

1.5 µL

1.5 mL

600 µL

Known variables 
You already know the volume of your organ bath. Add to the table the concentration of the provided stock solutions.

Organ Bath Volume (mL) Histamine Stock Solution (M)   Salbutamol Stock Solution (M)  

100.00 0.7e-6 1.75e-6

The doses of histamine and salbutamol you will add to the organ bath:

Final Bath Concentration (M) Dose of Histamine to Add (µL) Dose of Salbutamol to Add (µL)

1e-8 1.43e-3 5.71e-2

3e-8 4.29e-3 1.71e-3

1e-7 1.43e-4 5.71e-3

3e-7 4.29e-4 1.71e-4

1e-6 1.43e-5 5.71e-4

3e-6 4.29e-5 1.71e-5

1e-5 1.43e-6 5.71e-5

3e-5 4.29e-6 1.71e-6

1e-4 1.43e-7 5.71e-6

Known variables 
You already know the volume of your organ bath. Add to the table the concentrations of the provided stock solutions. 

Organ Bath Volume (mL) Acetylcholine Stock Solution (M)   Ipratropium Stock Solution (M)  

100.00 0.5e-6 1.68e-10

The doses of acetylcholine and ipratropium you will add to the organ bath:

Final Bath Concentration (M) Dose of Acetylcholine to Add (µL) Dose of Ipratropium to Add (µL)

1e-8 2.0e-3 5.95e-6

3e-8 6.0e-3 1.79e-7

1e-7 2.0e-4 5.95e-7

3e-7 6.0e-4 1.79e-8

1e-6 2.0e-5 5.95e-8

3e-6 6.0e-5 1.79e-9

1e-5 2.0e-6 5.95e-9

3e-5 6.0e-6 1.79e-10

1e-4 2.0e-7 5.95e-10

Histamine and salbutamol – Activity

What do you predict will be the response of the trachea to histamine application, then salbutamol application?

Histamine induces bronchoconstriction then the addition of salbutamol will inhibit the bronchial responsiveness to
histamine, producing bronchodilation.

Histamine and salbutamol – Analysis

Baseline Force (mN)

The e ects of histamine and salbutamol on contraction force.

Final Bath Concentration (M) Force with Histamine (mN) Force with Salbutamol (mN)

1e-8

3e-8

1e-7

3e-7

1e-6

3e-6

1e-5

3e-5

1e-4

Complete the graph by adding a title and labeling the axes.

Untitled Graph
Incomplete data in table
Unlabeled Axis

Unlabeled Axis
Force with Histamine (mN) Force with Salbutamol (mN)

What were the observed responses to the application of histamine, and then salbutamol? 

histamine will cause bronchoconstriction, while salbutamol will produce bronchodilation

Were your results di erent from your prediction? If so, how were they di erent?

No, they were not di erent. 

Histamine and salbutamol are physiological antagonists. When an asthma attack is treated with salbutamol, di erent
receptors are activated that oppose the e ects of the histamine, and reverse the symptoms of asthma.

Use the labels to order the signaling events that each drug triggers.  

Binds H1 receptors Binds β2 receptors Adenyl cyclase activated

Binds β2 receptors

Phospholipase C activated Adenyl cyclase activated Binds H1 receptors

Decrease intracellular Ca2+

Increase  intracellular Ca2+

Increase  intracellular Ca2+ Decrease intracellular Ca2+

Phospholipase C activated

Acetylcholine and ipratropium – Activity

What do you predict will be the response of the trachea to acetylcholine application, and then ipratropium?

Acetylcholine will cause bronchoconstriction. Ipratropium, being a muscarinic receptor antagonist will counteract

this e ect, resulting in bronchodilation.

Acetylcholine and ipratropium – Analysis

Baseline Force (mN)

The e ects of acetylcholine and ipratropium on contraction force.

Final Bath Concentration (M) Force with Acetylcholine (mN) Force with Ipratropium (mN)

1e-8

3e-8

1e-7

3e-7

1e-6

3e-6

1e-5

3e-5

1e-4

Acetylcholine EC50

Minimum response (mN)

Maximum response (mN)

50% response (mN)

EC50 [ACh] at 50% response (M)

Ipratropium IC50

Minimum response (mN)

Maximum response (mN)

50% response (mN)

IC50 [IPRA] at 50% response (M)

Complete the graph by adding a title and labeling the axes.

Untitled Graph
Incomplete data in table
Unlabeled Axis

Unlabeled Axis
Force with Acetylcholine (mN) Force with Ipratropium (mN)
What were the observed responses of the tissue to acetylcholine application, and then ipratropium? 

Acteylcholine will produce bronchoconstriction while ipatropium will cause bronchorelaxation

Were your results di erent from your prediction? If so, how were they di erent? 

No, they were not di erent. 

Describe the mechanism by which acetylcholine causes bronchial contraction, and also the mechanism by which
ipratropium causes bronchial relaxation. 

Acetylcholine will bind to m3 receptors, activating the Gq subunit of the G protein. Phospolipase C will then be
activated, converting PIP2 to IP3 and DAG. These will then increase intracellular calcium and therefore cause
bronchoconstriction. Ipratropium acts as an antagonist of the muscarinic acetylcholine receptor. This e ect produces
the inhibition of the parasympathetic nervous system in the airways and hence, inhibit their function. The function of
the parasympathetic system in the airway is to generate bronchial secretions and constriction and hence, the inhibition
of this action can lead to bronchodilation and fewer secretions. 

Theophylline – Activity

What do you predict will be the response of the trachea to salbutamol following histamine and theophylline application? 

Theophylline further enhance the bronchodilatory e ect that was already imposed by the salbutamol.

Calculate the dose of theophylline to add to the water bath. 

Final Bath Concentration (M) Organ Bath Volume (mL) Theophylline Stock Solution (M)  Dose to Add (µL)

5e-5 100.00 2.8e-5 1.79e-2

Theophylline – Analysis

Baseline Force (mN)

The e ect of the interaction of theophylline and salbutamol on tracheal contraction.

Final Bath Concentration (M) Force with Histamine (mN) Force with Salbutamol in the Presence of
Theophylline (mN)

1e-8

3e-8

1e-7

3e-7

1e-6

3e-6

1e-5

3e-5

1e-4

Complete the graph by adding a title and labeling the axes.

Untitled Graph
Incomplete data in table
Unlabeled Axis

Unlabeled Axis
Force with Histamine (mN) Force with Salbutamol in the Presence of Theophylline (mN)

What was the response of the trachea when theophylline was present with salbutamol and histamine? 

There was enhanced bronchodilation.

Were your results di erent from your prediction? If so, how were they di erent?

No, they were not di erent.

The combination of salbutamol and theophylline is an example of synergy. De ne the term synergy.

An interaction between two or more drugs that causes the total e ect of the drugs to be greater than the sum of the
individual e ects of each drug. A synergistic e ect can be bene cial or harmful.
Describe the mechanisms of how salbutamol and theophylline act to reduce contraction strength in the trachea. Explain
how they compliment each other to act in a synergistic way.

On a cellular level, it is known that theophylline inhibits mediator release from mast cells by decreasing destruction of
cyclic adenosine monophosphate whereas salbutamol binds to beta2 adrenergic receptors which increases CAMP that
leads to decreased intracellular calcium. Ultimately, both theophylline and salbutamol cause increased CAMP levels.
Therefore, the interaction of these two drugs might be explained ultimately on a cellular level by their ability to a ect
mediator release by acting on di erent biochemical pathways.

Summary

Describe in everyday language how salbutamol is used to treat acute asthma symptoms.

During acute asthma attacks, the airways are narrowed and lled with mucus that it is di cult for air to move in and
out. Salbutamol, usually given through an inhaler, o ers quick relief during acute asthma attacks by relaxing the
muscles of our airways and therefore helps us breathe more easily.

Popup - Notebook
 Re ection notebook:

This was a very stimulating activity. 

Popup - Table: known variables (Histamine and

Salbutamol)

Your known variables: 

Organ Bath Volume (mL) Histamine Stock Solution (M)   Salbutamol Stock Solution (M)  

100.00 7e-7 2e-6

Popup - Table: known variables (I and A)

Your known variables: 

Organ Bath Volume (mL) Acetylcholine Stock Solution (M)   Ipratropium Stock Solution (M)  

100.00 5e-7 2e-10