Antihistaminic activity of Ethanolic extract of Capparis moonii W. fruit.

 

Ms. Priya Gupta1, Dr. Vanita Kanase2*

1Department of Pharmacology, Oriental College of Pharmacy, Sector 2, Behind Sanpada Railway Station, Sanpada West, Navi Mumbai, Maharashtra 400705.

2HOD Pharmacology, Department of Pharmacology, Oriental College of Pharmacy, Sector 2,

Behind Sanpada Railway Station, Sanpada West, Navi Mumbai, Maharashtra 400705.

*Corresponding Author E-mail: vanita.kanase@gmail.com

 

ABSTRACT:

The purpose of the present work were intended to determine the antihistaminic activity of ethanolic extract of Capparis moonii W. fruits (EECM). Capparis moonii W.  had been historically used in the diagnosis of cough and asthma and so we undertook this study to validate scientifically using appropriate animal models. Antihistamine is considered to be helpful for the treatment of allergic, thus, the antihistamine activity of an ethanolic extract of Capparis moonii W. in the current work was evaluated. To determine the doses, acute oral toxicity tests were conducted. Clonidine and haloperidol that induced cataleptic effect in Swiss albino mice were evaluated for antihistaminic activity at the different doses of 50mg/kg, 100mg/kg and 200mg/kg, p.o. and the evaluation is also done on guinea pig ileum tissue. The ethanolic extract of Capparis moonii W. fruits (50, 100, 200mg/kg, p.o.) and chlorpheniramine maleate (i.p.,10mg/kg) significantly inhibited (****P<0.0001) clonidine induced catalepsy but the extract donot inhibit haloperidol-induced catalepsy and histamine-induced contraction in guinea pig ileum tissue preparation shows that ethanolic extract of Capparis moonii W. inhibited the contractile activity of histamine. The result of our work shows that the ethanolic extract possesses antihistaminic activity. It can be reported that flavonoid present in the extract may be important for an antihistaminic effect and therefore may have a role in the asthma treatment.

 

KEYWORDS: Antihistamine, Clonidine, Haloperidol, Catalepsy, Capparis moonii W., Guinea pig ileum.

 

 


INTRODUCTION:

Histamine is referred to common symptoms and allergic reactions. They are mostly comparable with histamine intolerance. Many typical reactions to this allergy can vary, which include migraine or headache, trouble with nasal congestion or sinus, fatigue, hives, stomach issues, abnormal menstrual period, nausea, and vomiting. Histamine is the compound for the immune response produced by the mastocyte. Antihistamines are a type of medication which is often used to manage a different variety of allergic problems in your body. Antihistamines are drugs used for the prevention of allergic rhinitis and allergies1,2.

 

Histamine is activated by Mast cells activation and basophils, via immunology and non-immunological mechanisms which Contribute to Allergic and Anaphylaxis reactions3. Catalepsy is a disorder of animal experiences long imposed posture before he regained his normal posture. Catalepsy is an indication of the extrapyramidal syndrome of medications inhibiting dopaminergic transmission or through the release of histamine into the brain4. Ayurveda, An Indian medicine method that identified many medicines for use in the diagnosis of asthma and allergy from indigenous natural sources5. Capparis moonii W. belonging to the Capparidaceae/Capparaceae family. Widely known as the Large Caper. Rudanti in Sanskrit and Waghati in Marathi. Capparis moonii Wight is distributed in Maharashtra, Goa, Karnataka, Tamil Nadu and it is frequently found in Konkan and Sri Lanka which grows normally in the hot climate. Capparis moonii Wight Fruits are sub-globose or ovoid. The fruit contains several large seeds. Capparis moonii W. fruits extract contains beta-sitosterol, l-stachyhydrin, rutin, as well as flavonoid, alkaloid, and glycoside6-8. Plants with flavonoids had been noted to have antihistaminic, antiallergic, and degranulation characteristics of the mast cells9. Family of Capparidaceae containing various species of Capparis and it is cultivated in the subtropical and tropical region of India; Its medicinal use had been studied in conventional medicinal systems.

 

Rudanti were traditionally used to provide asthma and cough relief10.

 

MATERIAL AND METHOD:

The dried fruits of Capparis moonii W. were collected from Hamidiya Dawakhana, Pydhony, Bhuleshwar, Mumbai-400003 in July 2019. Sample specimen voucher was submitted to Dr. Bindu Gopalkrishnan (Asst. Professor) from Mithibai College of Arts, Chauhan Institute of Science and Amruthben Jivanlal College of Commerce and Economics, Vile Parle (W), Mumbai- 400056.

 

The fruits were washed with tap water and shade dried at normal room temperature with the aid of circulating airflow using a fan. Then the powdered fruits plant of Capparis moonii Wight were extracted on soxhlet extractor using 90% v/v ethanol to obtained ethanolic extract of Capparis moonii Wight fruit. The extracts were evaporated to obtain the dry powder of extract. The percentage yield of ethanolic extracts was 7.09% w/w. These crude dried extracts were put in a suitable container and kept in refrigerator 4°C until use.

 

Qualitative Phytochemical Screening:

Preliminary chemical tests were carried out on ethanolic extract of Capparis moonii W. for determination of the presence of phytoconstituents like alkaloids, flavonoids, cardiac glycosides, saponins, steroids, tannins, terpenoids, and phenolic compounds.

 

Experimental Animals:

Either sex Swiss Albino Mice (25-25g) and Guinea Pig Male (300-400g) were used for the study. The Swiss Albino Mice were obtained from Bombay Veterinary College Parel, Mumbai-400012, and Guinea Pig were obtained from Bharat Serum and Vaccines Limited, Thane-400604. The use of these animals and the study protocols were approved by CPCSEA recognized institutional ethics committee of Oriental College of Pharmacy. Mice were kept at the animal house of Oriental College of Pharmacy, Sanpada, Navi Mumbai. The animals were allowed on a comfortable polypropylene cage with proper bedding followed by dark and light cycle at a steady temperature of 22 ± 2°C. They were provided with filtered water and pellets on a daily diet.

 

Acute Toxicity Study:

Acute oral toxicity studies were carried out for EECM as per the (Organization for Economic Cooperation and Development) OECD guideline number 42311.

 

Chemicals:

1.    Ethanolic extract of Capparis moonii W. (EECM) (50 mg/kg, 100mg/kg, 200mg/kg)

2.    Standard drug: Chlorpheniramine maleate (Cadistin) and Histamine.

3.    Drug to induce catalepsy: Clonidine (inj. Cloneon) and Haloperidol (inj. Serenace)

4.    Distilled water

 

METHODS:

A. Clonidine induced catalepsy in mice:

 Mice (Swiss Albino) were divided into 5 class, each containing 5 mice. Normal control (class 1) given a saline solution (10ml/kg) and a single dose of the extract (50, 100, 200mg/kg body weight) given to another class 3 to class 5. Class 2 mice received a standard dose of chlorpheniramine maleate (antihistamine) (10mg/kg, i.p.). One hour after drug consumption all classes received clonidine (1mg/kg s.c.) and the catalepsy period was determined at 0min, 15min, 30min, 60min, 90min, 120min, 150min, and 180 min12,13.

 

B. Haloperidol induced catalepsy in mice:

Mice (Swiss Albino) were divided into 5 class, each containing 5 mice. Normal control (class 1) given a saline solution (10ml/kg) and a single dose of the extract (50, 100, 200mg/kg body weight) given to another class 3 to class 5. Class 2 mice received a standard dose of chlorpheniramine maleate (antihistamine) (10mg/kg, i.p.). One hour after drug consumption all classes received haloperidol (1mg/kg i.p.) and the catalepsy period was determined at 0min 15min, 30min, 60min, 90min, 120min, 150min, and 180 min14,15.

 

Bar Study:

The bar method used for the measurements of catalepsy. In the bar study, the front paw of the animal was put on a horizontal bar 3 cm above and 5 cm parallel to the base, as an alternative. The period mice takes her front paw from the bar were noted.

 

C. Guinea pig ileum tissue preparation:

The male guinea pig weighing 300-400g were fasted overnight and then sacrificed, the ileum was excised and put in an organ bath with the prepared Tyrode solvent, which was periodically aerated at 37±0.5°C. The response curve for histamine were conducted in the plain Tyrode solvent and also in the Tyrode solvent containing Capparis moonii W. extract. The contractile responses were graphed to generate the histamine dose response curve in the absence and presence of Capparis moonii W. extract.16,17

Statistical Analysis:

The result were analysed with Prism Software by GraphPad (version 8.4.2). The results for each category will be expressed as the mean± SEM for each group. Statistical variations were evaluated using One-way variance analysis (ANOVA) accompanied by Dunnett's t-test. Where**** represents significant at p≤0.0001 as compared with Normal control.

 

RESULT:

A. Acute oral toxicity:

As suggested in OECD Guideline number 423, acute oral toxicity evaluation was conducted and the observation confirms that no toxic effects were observed in clinical criteria during an acute toxicity test of up to 2000mg/kg. This also indicates that the LD50 (lethal dose) of dried fruit of ethanolic extract of Capparis moonii W. is equivalent to 2000 mg/kg. After a detailed survey of numerous study papers and guided by the guide, doses were selected as 50 mg/kg, 100 mg/kg, and 200 mg/kg for study groups.

 

B. Qualitative phytochemical screening:

Table 1: Result of quantitative phytochemical analysis of powdered fruits of Capparis moonii W.

Sr. No.

Phytoconstituents

Ethanolic extract of the leaves Capparis moonii W.

1

Alkaloids

+

2

Flavonoids

+

3

Cardiac Glycosides

+

4

Saponins

+

5

Steroids

+

6

Terpenoids

-

7

Tannins and Phenolic compounds

+

8

Carbohydrates

-

9

protein

-

Present(+) Absent (-)              

 

The presence of flavonoids and the other components included in the extract corresponds significantly to the antihistaminic activity of the ethanolic extract of Capparis moonii Wight.

 


B. Clonidine induced catalepsy:

Table 2: Effect of Ethanolic extract of Capparis moonii W. fruits on inhibiting clonidine induced catalepsy in mice for a total time duration of 180 mins.

Duration of catalepsy (secs) mean±sem

GROUPS

0 MIN

15 MIN

30 MIN

60 MIN

90 MIN

120MIN

150 MIN

180MIN

Vehicle Control

(Normal saline)

0.183

27.08±

0.24

46.0±0.34

65.91±

1.12

84.0±

1.16

92.67±

0.67

96.25±

0.90

90.91±

2.21

Standard (Chlorpheniramine maleate)

1.18±

0.10

21.58±

1.52****

28.83±

1.47****

34.67±

1.40****

37.75±

1.71****

40.75±

1.63****

39.50±

1.85****

52.33±

1.07****

EETO 50mg/kg

(Low dose)

2.83±

0.35

27.75±

2.305****

34.83±

1.35****

42.41±

1.58****

53.83±

1.53****

68.33±

1.07****

76.50±

0.84****

79.58±

1.428****

EETO 100mg/kg

(Medium dose)

4.91±

0.15

32.08±

1.193****

38.83±

1.57****

42.10±

1.339****

52.18±

2.16****

68.25±

0.86****

71.16±

1.108****

71.917±

0.79****

EETO 200mg/kg

(High dose)

4.0±0.18

31.0±

1.34****

33.34±

1.59****

37.08±

0.95****

42.96±

1.68****

45.25±

1.30****

52.75±

1.64****

64.25±

1.27****

Values are the Mean ± SEM of n=6, **** represents significant at p≤0.0001 when compared to the normal control group.

 

C. Haloperidol induced catalepsy

Table 3: Effect of Ethanolic extract of Capparis moonii W. fruits on inhibiting haloperidol-induced catalepsy in mice for a total time duration of 180 mins.

Duration of Catalepsy (secs) Mean ± SEM

Groups

0 MIN

15 MIN

30 MIN

60 MIN

90 MIN

120 MIN

150 MIN

180 MIN

Vehicle Control (Normal saline)

4.41± 0.24

31.91±

1.58

53.0±

1.11

62.91±

2.25

78.75±

1.72

88.50±

1.072

102.083±

1.551

108.16±

0.33

Standard (Chlorpheniramine

maleate)

3.67± 0.211

31.67±

1.33****

40.167±

1.824****

59.16±

1.35****

66.58±

0.768****

78.66±

1.90****

90.83±

1.062****

92.00±

0.316****

EECM 50mg/kg

(Low dose)

7.33± 0.42

31.75±

1.48****

40.91±

1.48****

54.25±

1.82****

64.41±

1.446****

84.50±

1.42****

94.0±

1.58****

103.66±

2.171****

EECM 100mg/kg (Medium dose)

5.3± 0.188

31.76±

1.85****

40.58±

1.03****

54.46±

1.663****

65.58±

0.841****

80.33±

1.464****

91.33±

0.52****

102.08±

2.548****

EECM 200mg/kg (High dose)

6.58±

0.67

32.06±

2.12****

41.50±

2.661****

51.43±

0.552****

64.63±

1.648****

83.75±

1.564****

95.28±

1.22****

102.68±

0.358****

Values are the Mean ± SEM of n=6, **** represents significant at p≤0.0001 when compared to the normal control group.

 


Figure 2: Effect of Ethanolic extract of Capparis moonii W. fruits on inhibiting clonidine induced catalepsy in mice for a total time duration 180 mins represented in the bar graph.

 

Figure 3: Effect of Ethanolic extract of Capparis moonii W. fruits on inhibiting Haloperidol induced catalepsy in mice for a total time duration 180 mins represented in the bar graph.

Values are the Mean ± SEM of n=6, **** represents significant at p≤0.0001 when compared to the normal control group.

 

D. Guinea pig ileum tissue preparation

Table7: Effect of Ethanolic extract of Capparis moonii W. fruits on inhibiting contractions of smooth muscle present in guinea pig ileum induced by histamine.

Histamine concentration in 10 μg/ml

Histamine induced % maximum conc. (control) i.e. absence of EETO

DRC OF Histamine induced presence of EETO (Test)

0.1 ml

 14.83±1.25

 11.167±0.79

0.2 ml

 21.67±0.76

 21.167±0.307

0.4 ml

 31.83±0.703

 27.67±0.667

0.8 ml

 63.16±1.14

 45.0±1.125

1.6 ml

 87.5±0.764

 58.33±0.803

3.2 ml

 94.33±0.67

 66.33±1.308

Values are the Mean ± SEM of n=6 treatment

 

 

Figure 4: Effect of ethanolic extract of Capparis moonii W. fruits on inhibiting contractions of smooth muscle present in guinea pig ileum induced by histamine represented in the graph.

 

DISCUSSION AND CONCLUSION:

Clonidine and Haloperidol induced catalepsy:

Clonidine, an adrenergic alpha2-agonist, produces dose-dependent catalepsy in mice that had been primarily blocked by an antagonist of the histamine H1 receptor. It is reported in many studies that the extract which contains antihistaminic property or else mast cell stabilizing effect will suppress catalepsy which was induced by clonidine. Clonidine is responsible for the removal of histamine from mast cell which is important for different histamatic conditions.. The catalepsy produced by clonidine is regulated by H1 receptors through histamine. In this study we noticed that Catalepsy caused by clonidine was substantially prevented by treatment with EECM at various doses; thus, EECM has an antihistaminic activity.

 

On the opposite side, Haloperidol, a non-specific dopamine D2 antagonist that causes catalepsy is basically brought about by dopamine receptor blockage in the striatum. Haloperidol-induced catalepsy is inhibited by substances which boost dopamine transmission. This study was conducted to conclude that antihistaminic drugs did not suppress haloperidol generating catalepsy; while the catalepsy generated by clonidine is inhibited by antihistaminic drugs. Thus in this research analysis EECM inhibits the catalepsy caused by clonidine, which affirms the antihistamine property.

 

Isolated guinea pig ileum preparation:

General agonists, namely Ach, histamine, 5-HT, and bradykinine, are responsible for smooth contractile muscle behaviors. Isolated male guinea pigs ileum made of smooth muscles that are strongly sensitive to the contractions caused by histamine. Varied histamine doses were used for assessing the optimal contractile responses in the presence and absence of EECM. In this method, the DRC of histamine is shifted towards the right side that means ethanolic extract of Capparis moonii W. shows the presence of antihistaminic activity.

 

CONCLUSION:

The findings of the current study indicate that ethanolic extract Capparis moonii W. fruits can be beneficial for antihistaminic activity. Additional studies are suggested to classify the active phytochemicals and illustrate the mode of action.

 

ACKNOWLEDGMENT:

We are thankful for their guidance and help to our Principal Dr.(Mrs.)Sudha Rathod, Dr. (Mrs.) Vanita G. Kanase, as well as to the Department of Pharmacology, Oriental College of Pharmacy, Navi Mumbai.

 

AUTHORS’ CONTRIBUTIONS:

Dr. (Mrs.) Vanita Kanase guided with designing the study, making of protocol, and managed the work done. Priya Gupta performed the literature searches, performed the acute toxicity, models, phytochemical screening, and completed the manuscript writing.

 

CONFLICTS OF INTEREST:

We announce we do not have conflicting interests.

 

AUTHORS’ FUNDING:

We thank Oriental College of Pharmacy for funding the project.

 

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Received on 18.05.2020           Modified on 27.07.2020

Accepted on 07.09.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(8):4403-4407.

DOI: 10.52711/0974-360X.2021.00764