ANTISPASMODIC AND ANTHELMINTIC ACTIVITY OF NYCTANTHES ARBORTRISTIS LINN.HTML Full Text
ANTISPASMODIC AND ANTHELMINTIC ACTIVITY OF NYCTANTHES ARBORTRISTIS LINN.
SANJITA DAS*1, DINAKAR SASMAL2, SAUMYA PRIYA BASU1
Department of Pharmaceutical Technology, Noida Institute of Engineering and Technology*1, Greater Noida, (U.P.), India
Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi, (Jharkhand), India
Antispasmodic activity of the ethanolic extracts of different parts of Nyctanthes arbortristis Linn. was estimated using guinea pig ileum preparation against acetylcholine. Anthelmintic activity was tested following the method described by Kailashraj and Kurup, using earthworm (Pheretima posthuma). The extracts exhibited antispasmodic activity, which was less than that of piperazine citrate. The ethanolic extracts were found to have concentration-dependent paralytic activity, whereas its seeds and flowers showed lethal effect on the worms. It was also observed that the paralytic and lethal effects of respective ethanolic extracts were potentiated by the presence of atropine. It reveals that the anthelmintic activity of the extracts is due to the inhibition of motility by relaxing and depressing responsiveness to contractile action of acetylcholine.
Nyctanthes arbortristis Linn.
Nyctanthes arbortristis Linn. (Fam. Oleaceae), commonly known as Harsingar or Night Jasmine, is a common wild hardy large shrub or small tree. It is a native of India, distributed wild in sub-Himalayan regions and southwards to Godavari. It is also found in Indian gardens for ornamental purposes1-2. Its different parts are known to possess different pharmacological activities in Indian systems of medicines. Several phytochemical and pharmacological investigations have also been done on this plant4-12. The antispasmodic4 and anthelmintic1-3 activities of the leaves of Nyctanthes arbortristis L have been reported. The present study is carried out with the ethanolic extracts of its different parts like flowers, barks, seeds and leaves to study and confirm its anthelmintic activity.
MATERIALS AND METHODS:
Plant material: The flowers, barks, seeds and leaves of Nyctanthes arbortristis L. were collected from the gardens and forests of Orissa. The herbarium of the plant (CNH/I-I(20)/2005-Tech-II/254) was authenticated as Nyctanthes arbortristis (Fam.Oleaceae) from Botanical Survey of India, Kolkata. The dried leaves, barks and seeds were powdered coarsely and than were extracted successively with petroleum ether, chloroform and ethanol (90%) in soxhlet apparatus10-12. Its fresh flowers were extracted with ethanol (50%)10-11. The ethanolic extracts of leaves, barks, seeds and flowers were evaporated to dryness to get dark gummy masses, having yield value 14%, 12.5%, 26.5% and 13% respectively. The water soluble-portions of the extracts were subjected to further pharmacological screening.
Animals: Guinea pig weighing 400-600 g was obtained from the animal house of B.I.T., Mesra, Ranchi. It was kept under controlled environmental conditions allowing free access to food and water and acclimatized for at least a week before the commencement of the experiment. The Institutional Animals Ethics Committee (Registration No. 62/02/ac/CPCSEA) approved the experiments.
Antispasmodic activity: The antispasmodic activity was estimated using guinea pig ileum preparation against acetylcholine, used as the spasmodic agent13-14. Effect of the isolated compound on the concentration response curve of acetylcholine was observed. Then keeping the concentration of acetylcholine constant in the bath solution, responses were observed with the increasing concentrations of arbortristoside-A to estimate the amount, required to block the contraction that was produced by acetylcholine.
Anthelmintic activity: The anthelmintic activity was observed according to the method described by Kailashraj and Kurup (1962)15. Six earthworms, Pheretima posthuma of nearly equal size (8±1cm) were placed in each petridishes containing 15ml of normal saline and different dilutions (0.1%, 0.2%, and 0.5%) of the extracts and the standard piperazine citrate with normal saline. The time taken by the worms to become motionless, considered as to paralysis was recorded.
The lethal time was recorded by observing the time taken to become motionless on repeated application of external stimuli by pricking a pin. In the similar manner the experiment was repeated with all the extracts at different dilutions along with 0.1 ml atropine and the time taken by the worms for paralysis and the lethal time were recorded.
Statistical analysis: The results were subjected to statistical analysis, using ANOVA to determine the significance of the present study, where p<0.01 and p<0.001 were considered to be significant.
Antispasmodic activity: The ethanolic extracts of different plant parts of Nyctanthes arbortristis Linn. were found to reduce the concentration dependent responses produced by acetylcholine. The contractile response of 0.0002 mg of acetylcholine was inhibited by 90, 132, 120 and 72 mg of ethanolic extracts of the seeds, leaves, barks and flowers of Nyctanthes arbortristis Linn. Respectively, whereas piperazine citrate at 16 mg (Table 1).
Anthelmintic activity: The results are summarized in Table 2. The ethanolic extracts exhibited a concentration dependent paralytic effect. The seeds and
flowers showed to cause lethal effect at the concentration of 2 and 5 mg/ml, whereas the leaves and barks showed lethal effect on the worms at 5mg/ml only. Anthelmintic activity of Nyctanthes arbortristis L is less than that of the standard; Piperazine citrate. It was also observed that the paralytic and lethal effects of the ethanolic extracts of the seeds, barks and flowers were observed to be potentiated by the presence of atropine and so of the standard drug piperazine citrate. But the ethanolic extract of the leaves showed to cause lethality at higher concentration of 5mg/ml.
Table 2: Effect of ethanolic extracts of different plant parts of Nyctanthes arbortristis Linn. on the response obtained by 0.0002 mg of Ach on isolated guinea pig ileum preparation
Isolated guinea pig ileum preparation:
|2 ml of Ach + Drug treatment||Amount (mg)required to block the contraction produced by 0.0002 mg of acetylcholine|
|Atropine (1 m g/ml)||0.0003|
|Piperazine citrate (10mg/ml)||16|
|Flowers extract (60 mg/ml)
Barks extract (60mg/ml)
|Seeds extract (60 mg/ml)||90|
|Leaves extract (60mg/ml)||132|
Table 1: Anthelmintic activity of different parts of Nyctanthes arbortristis Linn
|Time in min.|
|For paralysis||For death|
|Without Atropine||With Atropine||Without Atropine||With Atropine|
|73.0 ± 0.408
65.25 ± 0.947
55.0 ± 0.817
55.0 ± 0.814
|18.5 ± 1.259
9.75 ± 0.354
7.75 ± 0.25
|166.25 ± 1.75
144.5 ± 1.051
111.25 ± 1.493
|15.25 ± 0.479
10.25 ± 0.629
3.5 ± 0.289
252.75 ± 1.53
236.25 ± 1.75
|22.25 ± 0.629
20.0 ± 0.408
16.0 ± 0.823
|94.75 ± 1.25
81.0 ± 1.732
|29.0 ± 1.354
23.75 ± 0.75
13.0 ± 0.812
262.0 ± 2.102
|37.25 ± 1.023
27.25 ± 0.25
20.25 ± 0.344
|143.25 ± 2.136
116.5 ± 1.553
88.5 ± 0.646
|6. 25 ± 0.479
4.25 ± 0.335
3.75 ± 0.479
242.75 ± 2.287
213.75 ± 1.75
|16.75 ± 0.479
9.0 ± 0.408
6.0 ± 0.408
|142.5 ± 0.5
115.5 ± 0.646
90.5 ± 2.102
|125.0 ± 3.536
120.0 ± 3.536
43.0 ± 2.0
266.25 ± 1.75
101.5 ± 3.67
Values are expressed as Mean ± SEM NAF, NAB, NAS and NAL: Ethanolic extracts of the flowers, barks, seeds and leaves of Nyctanthes arbortristis L respectively
The present study was undertaken on the antispasmodic and anthelmintic activity of the ethanolic extracts of different plant parts of Nyctanthes arbortristis Linn; on the basis of inhibition of contractile effect of acetyl choline by various dilutions of the ethanolic extracts, it initiated the study that the potentiation of the activity of the atropine on the motility of anthelmintics. Out of which flowers and seeds of this plant possess good antispasmodic activity. The ethanolic extracts of the seeds and flowers of Nyctanthes arbortristis L have more potent anthelmintic activity than that of the barks and leaves, but less than that of the standard drug piperazine citrate.
It has been observed that the ethanolic extracts of seeds, barks and flowers of Nyctanthes arbortristis L inhibited the contractile response of acetyl choline. Further, potentiation of the anthelmintic activity of atropine by the ethanolic extracts of different plant parts of the native plant might be due to inhibition of motility by relaxing and depressing responsiveness to contractile action of acetyl choline.
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SANJITA DAS*, DINAKAR SASMAL, SAUMYA PRIYA BASU
Department of Pharmaceutical Technology, Noida Institute of Engineering and Technology*, Greater Noida, (U.P.), India
04 January, 2010
17 January, 2010
24 January, 2010
01 February, 2010