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Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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.2014-15Department of Pharmacology

Supervised by:Dr. Umesh Kumar Gilhotra

Professor and PrincipalG. D. Memorial College of

Pharmacy

Rajasthan University of Health Sciences, Jaipur

2014-2015

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Introduction1

Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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

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

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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.

6Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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.

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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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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.

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Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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Drugs used in brochial AsthmaA. 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 - BeclomethasoneEvaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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Signs and Symptom5

1. Cough 2. Chest tightness3. Coughing after physical activity4. Whistling Sound while breathing5. Bronchoconstriction6. Airway hyper-reactivity

Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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Diagnosis of asthma7

SpirometryChest x-ray Physical examinationBlood tests and sputum studies

Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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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. 

Selection of plant

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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.

15Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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.

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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.

Reported activities8

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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 inexpensiveEvaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

18Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

• 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

19Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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)

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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.

21Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

Soxhlat extraction with Ethanol

Dried

Powdered

Soxhlat Extracted with Petroleum ether

Fruit of Citrullus colocynthis

PREPARATION OF RAW MATERIAL

Freeze

22Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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 +

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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.

24Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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

25Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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]

26Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

1 2 3 4 5 60

10

20

30

40

50

60

70

80

90

100

**

**

**

** **

**

**

**

**

****

Control Test Standard

-ve Molar concentration

% M

axim

um C

ontr

actio

nEffect 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.

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

Biochemicalparameters such as:

1.Appearance ofPreConvusive Dyspnea

(PCD)2.Precentage protectionoffered by drug in the

PreConvulsive dyspneaTime (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]

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Before 1Hr 3Hr 24 Hr0

10

20

30

40

50

60

70

80

90

100

Treatment

Pre

Conv

ulsi

ve T

ime

(PCT

)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.

29Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

GROUPS

(N =4)TREATMENT EVALUATION

PARAMETERS

I

(Control)Milk 4ml/kg,( s.c.).

Biochemical parameterssuch 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

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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;

Control Std CC-100 CC-2000

1000

2000

3000

4000

5000

6000

7000 6355.8

4805.66

59545607

Tota

l no

.of l

euco

cyte

s

Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

31Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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.

32Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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.

Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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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

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Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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

35Evaluation of Antiasthmatic activity of Citrullus colocynthis fruit.

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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