INTERNATIONAL MEDICAL UNIVERSITY

MALAYSIA

BACHELOR OF SCIENCES (HONS) PHARMACEUTICAL CHEMISTRY

General Pharmacology
BCH2122/ BBS2125/ BBT2122
PC218/ BM118 /MB118

Experiment 1: Part 1: Effect of Histamine on the Guinea Pig Ileum

Part 2: Effect of Mepyramine to Histamine in the Guinea
Pig Ileum

Student Name : Chia Ve Ka

Student’s ID : 00000027828
Date of Experiment : 2/5/2019
Due Date : 13/5/2019
Introduction

The aim of this experiment being conducted is to observe the effect of an

agonist, histamine on the guinea pig ileum preparation. The guinea pig ileum is
a part of the small intestine and contains receptors for many agonists. This
program stimulates an isolated preparation of the guinea pig ileum, a smooth
preparation exhibiting little spontaneous contractile activity which is
extensively used for pharmacological studies. Histamine acts on histaminic (H1)
receptors to cause contraction of the guinea pig ileum. The response depends on
the dose of the histamine. Competitive antagonist on histaminic receptors such
as mepyramine if presence, reduces the effect of histamine in the guinea pig
ileum. The antagonism of the mepyramine can be reduced if we increase the
dose of the histamine.

Procedure

Refers to laboratory manual.

Results

Stock Histamine Vol Added (mL) Final Bath Conc Response (g)
Solution (M) of Histamine (M)
10-8 0.1 1x10-10 0
-10
0.3 1x10 0
-10
0.5 1x10 0
10-7 0.1 1x10 -9
0.9575
-9
0.3 1x10 2.149
-9
0.5 1x10 3.361
10-6 0.1 1x10 -8
5.354
-8
0.3 1x10 9.526
-8
0.5 1x10 11.96
10-5 0.1 1x10 -7
13.08
-7
0.3 1x10 14.85
-7
0.5 1x10 15.31
10-4 0.1 1x10 -6
15.72
-6
0.3 1x10 15.82
-6
0.5 1x10 -
Table 1: Effect of Histamine on the Guinea Pig Ileum.

Stock Histamine Vol Added (mL) Final Bath Conc Response (g)
Solution (M) of Histamine (M)
10-6 0.1 1x10-8 0
-8
0.2 1x10 0
-8
0.3 1x10 0
-5 -7
10 0.1 1x10 0
-7
0.2 1x10 0.5960
-7
0.3 1x10 0.8696
-4 -6
10 0.1 1x10 1.798
-6
0.2 1x10 3.830
-6
0.3 1x10 5.413
-3 -5
10 0.1 1x10 8.080
-5
0.2 1x10 12.97
-5
0.3 1x10 13.00
Table 2: Effect of Mepyramine on the Guinea Pig Previously Treated with
Histamine.

Appendix I
Figure 1: Graph of the effect of histamine on the guinea pig ileum which is
curve at left and effect of mepyramine to histamine on the guinea pig ileum
which is curve at right.

Figure 2: Curve fits result of Figure 1.

Figure 3: The response when final bath concentration of histamine at 1x10-10M
to 5x10-9M.

Figure 4: The response when final bath concentration of histamine at 1x10-8M to

1x10-6M.
Figure 5: The response when final bath concentration of histamine at 5x10-6M to
3x10-5M.

F Figure 6: The response when final bath concentration is 1x10-8M to

concentration of 1x10-7M with the presence of mepyramine.
F
1
g
m
s0W
.3
ma
s
h
(M
ine
p
H)5
is.0
EW
a
s
h
(M
-7e
p
H)1
is.0
EW
a
s
h
(M
-6e
p
H)3
is.0
EW
a
s
h
(M
-6e
p
H)5
is.0
EW
a
s
h
(M
-6e
p
H)1
is.0
E
-5
M
Figure 7: The response when final bath concentration is 3x10-7M to
concentration of 5x10-6M with the presence of mepyramine.

Figure 8: The response when final bath concentration is 1x10-5M to

concentration of 5x10-5M with the presence of mepyramine.
Appendix II
From the graph of the concentration versus the response, obtain the following
information:

1. When histamine is given alone;

a. The threshold dose is 1x10-9M
b. The EC50 is 1.864x10-8M
c. The does producing the maximum response is 3x10-6M
d. The maximum response (Emax) is 15.82g

2. When histamine is given in the presence of mepyramine (1x10-8M);

a. The threshold dose is 3x10-7M
b. The EC50 is 1.203x10-5M
c. The does producing the maximum response is 5x10-5M
d. The maximum response (Emax) is 13.81g

3. From the formula: KB = XB/r-1

KB: Dissociation constant of mepyramine
XB: Concentration of mepyramine
r: [EC50(AA)/EC50(A)]
1 x 10−8 M
a. The KB = 1.203 x 10−5 M
1.864 x 10−8 M

= 1.549x10-11M
b. The pA2 value where pA2 = - log(KB) = 10.81
Discussion

1. Is the effect of histamine on guinea pig ileum a graded or quantal

response?
The effect of histamine on guinea pig ileum is a graded response.

2. What receptor is stimulated by histamine?

H1-receptors is stimulated by histamine [1].

3. Was the antagonism by mepyramine full reversible by increasing the dose

of histamine? How?
Yes, mepyramine effect on receptor is fully reversible when dose of histamine is
increased. Mepyramine is competitive antagonist to histamine. Additionally, one
receptor can only be bound by one type of ligand at a time. Thus, increase in
concentration of histamine increase the probability for histamine to bound to
receptor.

4. What did you observe about the slopes of the dose-response curves of
histamine alone and in the presence of mepyramine?
Sigmoid curve was obtained when dose-response for histamine and with the
presence of mepyramine was considered. Based on figure 1, curve plotted
showed that response increased when histamine dosage increased. However
with the presence of mepyramine, the curve shifted to the right.

5. What kind of antagonism is this?

Reversible competitive antagonism is the kind of antagonism of mepyramine.
Conclusion
In part I, histamine was the only agonist added into organ bath and its threshold
dose to stimulate response was 1x10-9M. EC50 value obtained was 1.864x10-8M
and dose that produce maximum response at 15.82g was 3x10-6M.
In part II, 1x10-8M of mepyramine was added into reservoir while histamine into
organ bath, its threshold dose to stimulate response was 3x10-7M. EC50 value
obtained was 1.203x10-5M and dose that produce maximum response at 13.81g
was 5x10-5M.

State what you have learnt from this experiment:

I had learnt to operate organ bath simulation software to perform an experiment
with it. I had also learned on using biograph software to plot curves in order to
obtain EC50 reading.

Why do you as Pharm Chem used to do this experiment?

This experiment allows me to realise the importance of dose-response
relationship which could be useful when formulating a drug in the future.
Mechanism of drug and receptors from organ bath simulator was also
understood which helped in the understanding that different drug will target
different receptors.

Reference
1. Parsons M, Ganellin C. Histamine and its receptors. British Journald of
Pharmacology [Internet]. 2009 [cited 12 May 2019];147(S1):S127-S135.
Available from:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1760721/