1. The document evaluates the antiasthmatic activity of Citrullus colocynthis fruit in 3 experiments. 2. In the first experiment, the ethanolic extract of C. colocynthis fruit antagonized histamine-induced contractions in isolated goat tracheal preparations, suggesting antihistaminic effects. 3. In the second experiment, the extract prolonged the preconvulsive time in histamine-induced bronchoconstriction in guinea pigs, indicating bronchodilating effects. 4. In the third experiment, the extract decreased total leukocyte count in mice in a milk-induced leucocytosis model, demonstrating potential adaptogenic activity. 5. The results suggest

1. 1
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

2. Evaluation of Antiasthmatic activity of
Citrullus colocynthis fruits
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 2
Submitted by:
Chandan Genwa
M.Pharm (IV) sem.2014-15
Department of Pharmacology
Supervised by:
Dr. Umesh Kumar Gilhotra
Professor and Principal
G. D. Memorial College of
Pharmacy
Rajasthan University of Health Sciences, Jaipur
2014-2015

3. Introduction1
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 3
 Asthma is a chronic inflammatory respiratory disease characterized by
periodic attacks of wheezing, shortness of breath and a tight feeling in
the chest. A cough producing sticky mucus is symptomatic. The
symptoms often appear to be caused by the body's reaction to a trigger
such as an allergen (commonly pollen, house dust, animal dander).
Asthma is a problem in the airways due to multiple factors.
Asthma

4. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
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Asthma is a chronic inflammatory lung disease that can cause
repeated episodes of cough, wheezing and breathing difficulty.
In addition, excess mucus production occurs in the airway.
Airway Obstruction
Airway Hyper responsiveness
 Airway Inflammation

5. Classifying Asthma
 Over the years, there have been several ways of classifying asthma and of
distinguishing one form of the disease from another
1 Allergic Asthma: Allergic asthma is triggered by allergic-substances capable of
causing an allergic reaction. Pollutants, wood dust, smoke, irritants, chemicals,
viral infection, bacteria, stress, emotion, and exercise are other frequently
diagnosed causes.
2 Intrinsic Asthma: intrinsic asthma include respiratory irritants such as
perfumes, smoke and cold air, upper respiratory infections .
Intrinsic asthma tends to be less responsive to treatment than allergic asthma.
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 5

6. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 6
 Exercise Asthma: Exercise induced asthma can affect anyone at any age and
may be attributed to the less of heat and moisture in the lungs that occurs with
strenuous exercise
 Occupational Asthma: Occupational asthma occurs directly as a result of
breathing chemical fumes, wood dust, or other irritants over long periods of
time. An estimated 15 percent of asthmatics have occupational asthma.

7. Causes of asthma3
 Allergens
• Animal dander
• Dust
• Pollen from trees and
grass
 Irritants
• Smoke
• Air pollution
• Cold air
 Other
• Medicine
• Some chemicals
• Infections
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
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8. Pathogenesis of Asthma4
 In asthma, constriction of the airways occurs due to bronchial inflammation.
 Bronchoconstriction is the narrowing of the airways in the lungs due to the
tightening of surrounding smooth muscle. Bronchial inflammation also
causes narrowing due to edema and swelling caused by an immune response
to allergens.
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 8

9. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
9

10. Drugs used in brochial Asthma
A. Bronchodilators;
 1. β2 Sympathomimetics
Terbutaline, Salbutamol.
 2. Methylxanthines
Theophylline , Aminophylline
3 Anticholinergics.
Ipratropium bromide , Tiotropium
B. Leukotriene antagonists
Montilukast, Zafirlukast
C. Mast cell Stablizers
Ketotifen.
D . Corticosteroids
Inhalation - Beclomethasone
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
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11. Signs and Symptom5
1. Cough
2. Chest tightness
3. Coughing after physical activity
4. Whistling Sound while breathing
5. Bronchoconstriction
6. Airway hyper-reactivity
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 11

12. Diagnosis of asthma7
 Spirometry
 Chest x-ray
 Physical examination
 Blood tests and sputum studies
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
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13. The selection of plant was based on ethno medical/ tribal
information and literature surveys. The plants selected was
Citrullus colocynthis fruits as it was reported by tribal’s to be
the best for Asthma.
13
Selection of plant

14. Review of literature8
 Plant name : Citrullus colocynthis.
 Family: Cucurbitaceae.
 Synonyms:
 English – Colocynth
 Hindi – Tumba,Indrayan
 Panjabi – Ghurunba
 Scientific classification:
 Kingdom : Plantae
 Division : Spermatophytina
 Order : Cucurbitales,
 Family : Cucurbitaceae,
 Genus : Citrullus,
 Species : Citrullus colocynthis
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 14

15. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 15
Chemical Constituents:9
Carbohydrates, Phytosterols,Protein, and Flavonoids.
Colocynthoside A&B.
Traditional medicinal uses;10
The fruits are bitter, acrid, Carminative, Purgetive, Asthma, Ulcer,
Jaundice, Antihelminthic.

16. Sr.
no.
Activity Parts used
1 Antibacterial activity Leaves
2
Anti-inflammatory and Analgesic
activity
Fruits
3.
Antidiabetic activity fruits
4. Antimicrobial activity Fruits
5.
Antiallergic activity
Leaves
6. Hepatoprotective activity Roots
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 16
Reported activities8

17. Need of Present Study
 The present day medical management of asthma is either
costly or not without side effects. Hence, the search for
antiasthmatic drugs from natural sources has assumed
greater importance because they are effective with fever
side effect and are also inexpensive
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 17

18. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 18
• The extensive review of literature on Citrullus colocynthis.
• Collection of plant/preparation of herbarium sheets of the plant Citrullus
colocynthis.
• Extraction of the fruits of Citrullus colocynthis. Using polar and nonpolar
solvents.
• Detailed phytochemical investigation/tests of the plant extract.
• Toxicity study of ethanolic extract of Citrullus colocynthis.
• Evaluation of In vivo Antiasthamatic activity of plant extract by
Histamine induced Bronchoconstriction in Guinea pig model.
PLAN OF WORK

19. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
19
Plant Identification
The identification and authentication of the plant
was carried out by Dr. S. L. Meena. Scientist ‘C’ &
in Charge Botanical Survey of India, Jodhpur (Raj.)
(No.BSI/AZRC/I.I2012/TECH/2014-2015-(PI.ID/174)

20. Approval from IAEC/CPCSEA
The protocol of the animal experiments involved in this
research work has been approved by IAEC/CPCSEA
constituted for this purpose
Ref. No. :GDMCP/2014-15/244(F)
Regd. No.1491/PO/a/11/CPCSEA
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 20

21. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 21
Soxhlat extraction with Ethanol
Dried
Powdered
Soxhlat Extracted with Petroleum ether
Fruit of Citrullus colocynthis
PREPARATION OF RAW MATERIAL
Freeze

22. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 22
Phytochemical Analysis of Citrullus colocynthis Fruit
extract12
S. No
Test for Plant Constituents
Citrullus
colocytnhis
fruits Extract
with:
1 Test for Carbohydrates +
2 Test for Proteins and amino acids +
4 Test for fixed oils and fats -
5 Test for alkaloids +
6 Test for Glycosides -
7 Test for Flavoniods +

23. AIM OF WORK
The aim of present work was to evaluate the antiasthmatic activity of
Citrullus colocynthis fruit extract in following laboratory animal models:
• Isolated goat tracheal chain preparation.13
• Histamine induced Bronchoconstriction in Guinea pig.14
• Milk -induced Leucocytosis.15
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 23

24. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
24
ANIMALS :
Dunkin Heartly Guinea pigs
Swiss Albino mice
SELECTION OF PLANT
EXTRACT :
Citrullus colocynthis fruits ethanolic
extract
( CC Extract)
SELECTION OF DOSES :
100mg/kg,200mg/kg (p.o.)
for mice.
100 mg/kg,200mg/kg(p.o.)
for guinea pigs.
Experimental methods and results

25. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
25
Treatment
D.R.C. of histamine (30μg/ml) in absence of test and std drug.
D.R.C. of histamine (30 μg/ml) in presence of ethanolic extract of
Citrullus colocynthis fruits (100μg/ml).
D.R.C. of histamine (30 μg/ml) in presence of Chlorpheniramine
maleate (1 μg/ml).
1. Isolated goat tracheal chain preparation. [13]

26. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
26
**
**
**
**
**
**
**
**
**
**
**
0
10
20
30
40
50
60
70
80
90
100
1 2 3 4 5 6
%MaximumContraction
-ve Molar concentration
Control Test Standard
Effect of Citrullus colocynthis fruits(100 µg/ml) extract on histamine
induced contraction of isolated goat tracheal chain preparation.
Data are expressed as Mean ± S.E.M. Where, n= 5. Statistical analysis done by using Student’s
‘t’-test.**p<0.01, significantly different from control.

27. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 27
GROUPS
(N =4)
TREATMENT EVALUATION PARAMETERS
I
(Control)
Phosphate buffered saline
(1 ml/kg, p.o.)+ 0.2 % histamine
aerosol
Biochemical
parameters such as:
1.Appearance of
PreConvusive Dyspnea
(PCD)
2.Precentage protection
offered by drug in the
PreConvulsive dyspnea
Time (PCT)
II
(Standard)
Chlorpheniramine maleate
(2 mg/kg, i.p.) +0.2 % histamine
aerosol.
III
(CC-100)
(Citrullus colocynthis 100 mg/kg,
p.o.)+ 0.2 % histamine aerosol.
IV
(CC-200)
(Citrullus colocynthis 200mg/kg,
p.o.)+ 0.2 % histamine aerosol.
2. Histamine induced Bronchoconstriction in Guinea pig:[14]

28. 0
10
20
30
40
50
60
70
80
90
100
Before 1Hr 3Hr 24 Hr
PreConvulsiveTime(PCT)
Treatment
Effect of ethanolic extract of Citrullus colocynthis fruits on
histamine induced Bronchoconstriction in guinea pigs
Data are expressed as Mean± S.E.M. Where, n= 4. Statistical analysis done by ANOVA followed by
Dunnett’s test, where *p<0.05,**p < 0.01 when group II, III,and IV were compared with group I.
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 28

29. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 29
GROUPS
(N =4)
TREATMENT
EVALUATION
PARAMETERS
I
(Control)
Milk 4ml/kg,( s.c.).
Biochemical parameters
such as:
Difference in no. of
leukocytes count.
II
(Std)
Dexamethasone (50mg/kg) +Milk
(4ml/kg s.c.)
III
(CC-100)
(Citrullus colocynthis fruits 100 mg/kg,
p.o.)+ Milk 4ml/kg,( s.c.).
IV
(CC-200)
(Citrullus colocynthis 200 mg/kg,
p.o.)+ Milk 4ml/kg,( s.c.).
3. Milk-Induced Leucocytosis in Mice:15

30. Data are expressed as Mean± S.E.M. Where, n= 5,
Statistical analysis done by ANOVA followed by Dunnett’s test, where *p<0.05,**p < 0.0001 when group II,
III, and IV were compared with group I.
Effect of ethanolic extracts of Citrullus colocynthis fruits on
milk induced Leucocytosis in mice;
6355.8
4805.66
5954
5607
0
1000
2000
3000
4000
5000
6000
7000
Control Std CC-100 CC-200
Totalno.ofleucocytes
Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 30

31. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 31
Discussion
 Isolated goat tracheal chain preparation:
H1 Receptor Histamine
 Ethanolic extract of Citrullus colocynthis fruits has
antagonized the histamine induced contractions on goat tracheal
chain preparation.
 The stimulation of H1 receptor causes contraction of bronchial
smooth muscle.
Thus, ethanolic extract of Citrullus colocynthis fruits may possess
antihistaminic (H1-receptor antagonistic) action and thus may contribute in the
management of asthma.

32. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
32
 Histamine induced bronchoconstriction in guinea pigs:
Histamine when inhaled causes hypoxia , Which leads to
convulsion in guinea pigs
It causes very strong smooth muscle contraction.
leads to severe bronchoconstriction in the guinea pigs that can
causes hypoxia and death.
CC act as bronchodialator due to anti histaminic activity which
indicated by the increase in PCT (preconvulsive time). Thus may contribute in
the management of asthma.

33. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
33
 Milk induced Leucocytosis in mice:
Leucocytosis:
 After parenteral administration of milk there is increase in total leukocyte count.
Condition can be normalized by administration of an adaptogenic drug.
An important feature of the adaptogens is their capacity to increase organism
resistance to various adverse effects of a physical, chemical and biological
nature
Treatment with ethanolic extract of Citrullus colocynthis fruits showed
significant decrease in total leukocyte count. This effect may be due to its
adapatogenic activity.
So it may contribute in the management of asthma

34. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.
34
Ethanolic extract of Fruits of Citrullus colocynthis fruits may
possesses antiasthmatic activity which may be due to :-
antihistaminic activity, bronchodilating activity, adaptogenic
activity, anti-inflammatory activity (inhibition of antigen antibody reaction)
Thus Citrullus colocynthis fruits may be used in the management of
asthma.
Conclusion

35. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 35
Future Scope
Further study regards to isolation, purification, mechanisms and
pharmacological screening of the active principles present in Ethanolic
extract of Citrullus colocynthis fruits needs to be done and their exact
mechanism of action and clinical efficacy in the management of asthma
also need to be find out.

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4 Jarjour NN, Kelly EAB. Pathogenesis of asthma. Medical clinics of North America 2002; 86: 925-
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5 Suralkar Anupama A , Jadav Asha S.Antiasthmatic and Bronchodialating activity of fruits berries of
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37. 8 Borhade P, Deshmukh T, Patil V and Khandelwal K. Review on Citrullus colocynthis. Int J
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38. Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit. 38
13.Kulshreshtha S, Misra SS, Sharma AL, et al. Response of the goat trachea to
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39. Evaluation of Antiasthmatic activity of Citrullus
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