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Journal of Medicinal Chemistry and Drug Discovery

ISSN: 2347-9027 www.jmcdd.com Page 14

Available online athttp://www.jmcdd.com

October - November, 2014, Vol. 4, pp 14-25

ISSN: 2347-9027

Research Article

ANTI-DIARRHOEAL AND LEARNING AND MEMORY ENHANCING ACTIVITY OF

AERIAL PARTS OF BASELLA ALBA

Komati Pavani

Department of Pharmacology, St.Marys College of Pharmacy, Secunderabad, Telangana

Email id: [email protected]

ABSTRACT

The plant Basella Alba (Basellaceae) is a small ever green and possesses a number of medicinal properties. The purpose of the present study was

to evaluate the anti diarrhoeal activity of the ethanolic and aqueous extracts of the aerial parts of Basella Alba, by using castor oil induced

diarrhea model. The ethanolic and aqueous extracts of aerial parts of this plant at graded doses was investigated for anti diarrhoeal activity in

terms of reduction in the rate of defaecation and consistency of faeces in Castor oil induced diarrhea. Effect was further evaluated on

gastrointestinal motility test with charcoal meal and Learning and Memory enhancing activity also evaluated by using piracetam drug as a

standard. The aerial parts extract showed significant inhibitory activity against castor oil induced diarrhea and also showed nootropic activity.

There was significant reduction in gastro intestinal motility by the charcoal meal test in rats. The results obtained by this study substantiate the

anti diarrhoeal and nootropic effects of the ethanolic or aqueous extract.

KEY WORDS: Diarrhoeal activity, AEBA, EEBA, Castor oil, gastrointestinal motility.

INTRODUCTION

Diarrhoea is the production of soft or watery stools by increased frequency of bowel movements. It may be defined

as the passage of more than 300ml of liquid faeces in 24 hours. This results in fluid and electrolyte loss that may

lead ultimately to death, particularly in young children. Pain, urgency, perianal discomfort and incontinence often

accompany it1. Diarrhoea is caused by either various infectious agents or chemical agents.Nootropics(memory

enhancers) are drugs or neutraceuticals, or functional foods that are purported to improve mental functions such as

cognition, memory, motivation, attention, intelligence and concentration2-4.
http://www.jmcdd.com/http://www.jmcdd.com/http://www.jmcdd.com/http://www.jmcdd.com/


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Basella Alba commonly known as Indian spinach, belonging to the family Basellaceae. The aerial part(leaves, stem)

of the plant serve as edible plant(vegetable) in many parts of the world5. The leaf juice is a demulcent, used in casesof dysentery. Stem and leaves are used as mild laxative, diuretic and antipyretic. Also used in constipation, poultice

for sores, urticaria and gonorrhea. It is also used in poultice local swellings, intestinal complaints etc6. Present study

was conducted to investigate the antidiarrhoeal effect of Basella alba.

MATERIALS AND METHODS

Collection and identification of Plant material

Collection and drying of plant material: The aerial parts of Basella alba were collected from village areas of Siddipet

and authentified by botanist Dr. Rasingham BSI, Hyderabad and the voucher samples are kept in the BSI herbarium

for reference (BSI/DRC/12-13/Tech. /436).

Preparation of different extracts (petroleum ether, chloroform, ethanolic and aqueous extracts)7:

The stems of Basella alba were dried in shade at room temperature then made into a fine powder . Then the powder

was extracted with petroleum ether, chloroform, ethanol and water successively by soxhlet apparatus, maintained at

50oC to get extracts. These extracts were stored in airtight containers in a refrigerator below 10oC. The extracts

were examined for their colour and consistency. Their percentage yield was calculated with reference to air-dried

powder sample used for the extraction.

Animal selection:

For Antidiarrhoeal activity Wistar albino rats of either sex weighing between 150-200g and adult Swiss albino mice

(20-25g) of either sex, obtained from Srivenkateshwara enterprises, Bangalore were selected. The animals were

acclimatized to standard laboratory conditions (temperature 252oC) and maintained on 12 hours light; 12 hours

dark cycle. They were fed with ad libitum. Before performing the experiment the ethical clearance was obtained

from institutional animal ethics committee (IEAC). IAEC No.769/2010/CPCSEA.

Acute toxicity study:

The acute toxicity of Basella alba extract was determined by using albino mice (20-25g) those maintained under

standard husbandry conditions. The animals were fasted 12h prior to the experiment and acute oral toxicity as per

OECD guideline no. 420 was determined.


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Animals were administered with single dose of 2000 mg/kg extract and observed individually with special attention

given during the first 3 hours and for a total of 14 days. During this period the mortality and/or the moribund statusof the animals were noted8.

Anti-diarrhoeal activity:

Castor oil-induced diarrhoea9:

In the present study albino mice of either sex weighing 20-25 gm were used. They are divided in to six groups of 6

animals each. The various groups were treated as follows:

Group -1: Control (Castor oil 0.5 ml)

GroupII: Standard (Loperamide + castor oil 0.5 ml p.o.)

GroupIII: Ethanolic extract (200 mg/kg p.o. + castor oil 0.5 ml p.o.)

GroupIV: Ethanolic extract (400 mg/kg p.o. + castor oil 0.5 ml p.o.)

GroupV: Aqueous extract (200 mg/kg p.o. + castor oil 0.5 ml p.o.)

GroupVI: Aqueous extract (400 mg/kg p.o. + castor oil 0.5 ml p.o.)

Subsequently, the vehicle, loperamide and extracts were administered, orally to groups of 6 mice 60 min before theadministration of the castor oil. The following parameter were determined, the time elapsed between the

administration of the cathartic agents and the excretion of the first diarrhoeic faeces (wet faeces that leaves a hallo

on the filter paper); the total number faeces as well as the number of diarrhoeic faeces excreted by the animals in 4

hrs; and the total weight of diarrhoeal stool in the period of time.

Gastrointestinal motility test10:

In the present study albino mice of either sex weighing 20-25 gm were used. They are divided in to 6 groups of 6

animals each. The various groups were treated as follows:

GroupI: Control (Charcoal meal 0.5 ml p.o.)

GroupII: Standard (Atropine sulphate i.p. + Charcoal meal 0.5 ml p.o.)

GroupIII: Ethanolic extract (200 mg/kg p.o. + Charcoal meal 0.5 ml p o.)

GroupIV: Ethanolic extract (400 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)


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GroupV: Aqueous extract (200 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)

GroupVI: Aqueous extract (400 mg/kg p.o. + Charcoal meal 0.5 ml p.o.)

Subsequently, after 30 min each individual mouse was administered with 0.5 ml of a 5% charcoal suspension in a

10% aqueous suspension of tragacanth mucilagel was administered p.o. to each animal.

After 30 min of this treat, each rat was sacrificed and intestinal distance moved by the charcoal meal from pylorus to

cecum. The percentages of distance travelled by the charcoal meal in ratio to the intestinal length and percentage of

inhibition was calculated by using following formula.

Distance travelled by the charcoal meal % travelled = x 100

Total length of small intestine

(Total length of the small intestineDistance travelled by the charcoal meal) % of inhibition = x 100

Total length of small intestine

Learning and Memory enhancing activity11-21:

Experimental Procedure: Laboratory model for testing learning and memory:

Elevated plus maze test: The elevated plus maze was described as tool for testing memory by the investigator

working in the field of psychopharmacology. Elevated plus maze served as extroceptive behavioral model to

evaluate learning and memory in rats. The elevated plus maze consists of two open arms and two closed arms

(50cmx10cmx40cm) with an open roof arranged so that the two arms are opposite to each other. The maze was

elevated to a height of 50 cm.

Model A: Effect of extracts on Transfer ratio in Diazepam induced amnesia:

Mice were divided into 7 groups consisting of 6 animals per group. Group-1 treated with Normal control (Distilled

water 10ml/kg, p.o) only once daily for 14 days, Group-2 treated with Diazepam (5mg/kg, i.p) alone on first day

only and after 45 mins, TL was recorded on EPM and retention (memory) of learned task was observed 24 hrs later.

The TL was calculated as described above. Group-3 treated with standard drug piracetam (200 mg/kg, p.o). Groups


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3, 4 and 5, 6 were treated orally with different doses EEBA (200 & 400 mg/kg) and AEBA (200 & 400 mg/kg)

respectively only once daily for 14 days.

On the 14th day, 90 minutes of treatment of last dose, TL is recorded. Twenty four hours later i.e., on 15th day

transfer latency was again recorded.

Statistical analysis:

All results will be expressed as mean SEM from 6 animals. Statistical difference in mean will be analyzed using

one-way ANOVA (analysis of variance) followed by Post hoc test (Dennettst test). P< 0.05*, 0.01** and

0.001*** will be considered as statistically significant.

RESULTS & DISCUSSION

The % yields of Aqueous and Ethanolic etracts are 10.0 and 8.0 respectively. The preliminary phytochemical

analysis of petroleum ether, chloroform, ethanolic, and aqueous extracts of aerial parts of Basella alba revealed the

presence of various phytoconstituents (table-1)

Acute toxicity test

The mice treated with EEBA and AEBA at a dose of 2000 mg/kg, p.o. exhibited normal behaviour, without any

signs of passivity, stereotypy and vocalization. Their motor activity and secretory signs were also normal and no

sign of depression. EEBA and AEBA even up to the dose level of 2000 mg/kg body weight did not produce any

behavioural symptoms or mortality. So 1/10th and 1/5th doses of (maximum dose tested for each extract) were

selected as medium and high doses and were used in the present study to explore anti diarrhoeal and learning and

memory enhancing activities.

Anti-diarrhoeal activity:

Castor oil-induced diarrhoea:

Effect of ethanolic and aqueous extracts of aerial parts of Basella alba on castor oil induced diarrhoea:

All the parameters of castor oil induced diarrhoea (total no. of faeces, on set time of diarrhea, no. of wet feces and

total weight of wet feces (fig.1-3)) were reduced by treatment of ethanolic extracts and aqueous extracts of aerial

parts of Basella alba where as the time required for diarrhoeal episode have been significantly reduced by the above

mention extracts as well as the standard drugs (loperamide). Between the two extracts ethanolic extract was found to

be more potent (table-2).


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Table1:Phytochemical evaluation of different extracts of Basell a alba

S.No. Tests Petroleum ether Chloroform Ethanol Water1 Alkaloids -ve -ve +ve +ve

2 Carbohydrates -ve -ve +ve +ve

3 Flavonoids -ve -ve +ve +ve

4 Saponins -ve -ve +ve +ve

5 Sterols +ve +ve +ve +ve

6 Tannins -ve -ve +ve +ve

7 Glycosides -ve -ve +ve +ve


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Table No.2: Effect of EEBA & AEBA on castor oil induced diarrhoea

Treatment Dose (mg/kg) 0nset time of

diarrhoea

(min)

Total

number of

faeces

Number of

wet faeces

Total weight

of wet faeces

(mg)

Control Distilled water 103.51.8 9.161.13 6.330.80 35353.89

Standard Lopera-mide(3mg/kg)

203.50.4** 4.230.73** 3.450.61** 169.166.83**

EEBA 200 mg/kg 140.51.8* 5.500.56* 4.150.49* 198.535.44*

EEBA 400 mg/kg 195.51.9** 5.021.07** 3.631.01** 178.2314.04**

AEBA 200 mg/kg 132.50.8 6.030.77* 5.031.05 223.1636.91

AEBA 400 mg/kg 135.56.6* 5.830.54* 4.531.05* 221.6614.87*

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P < 0.01, When compared tocontrol group. Standard drug: Loperamide (3mg/kg).

Fig.No.1: 0nset time of diarrhoea in castor oil induced diarrhea

0

50

100

150

200

250

Timeinmin

Groups

0nset time ofdiarrhoea (min)


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Fig.No.2: Total number of faeces in castor oil induced diarrhea

Fig.No.3: Total weight of wet faeces in castor oil induced diarrhoea

Gastrointestinal motility:

Effect of ethanolic and aqueous extracts of aerial parts of Basell a albaon Gastrointestinal motility:

Pre-treatment with ethanolic and aqueous have

reduced the GIT motility in a dose dependent manner by the reduction in the movement charcoal meal.

Between the two extracts ethanolic extract was found (table-3) to be most potent .

0

2

4

6

8

10

12

Numberoffaeces

Groups

Total

faeces

wet faeces

0

200

400

600

Totalweightof

wetfaeces

Groups

Total weight of wetfaeces


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ISSN: www.jmcdd.com Page 22

Table No.3: Effect of EEBA & AEBA on Gastrointestinal motility

S.NO Treatment Dose(mg/kg)Distance traveled by charcoal meal

with SEM (%)

1Control Distilled water 91.254.76

2 StandardAtropine sulphate(0.1 mg/kg)

56.102.30**

3 EEBA 200 mg/kg 64.963.56*

4 EEBA 400 mg/kg 61.682.38**

5 AEBA 200 mg/kg 77.054.58

6 AEBA 400 mg/kg 72.834.0*

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P < 0.01, When compared tocontrol group.Standard Drug: Atropine sulphate (i.p)

Fig.No.4: Distance travelled by charcoal meal in Gastrointestinal motility

0

20

40

60

80

100120

Distancetraveledby

charcoalmealwithSEM

(%)

Groups

Distance travelled by

charcoal meal


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Learning and memory enhancing property:

The ethanolic and aqueous extracts of aerial parts of Basella alba were studied for the learning andmemory enhancing properties. The results explain the transfer latencies (TL) of the various drug treated

groups i.e., control, toxic control, standard, ethanolic and aqueous extracts of the extracts of aerial parts ofBasella alba(table-4).

Table No 4: Nootropic effect of aerial parts of Basell a albain mice with EPM model.

Group

No.Treatment Dose (per Kg)

Transfer Latency

14 day 15 day

I. Normal control 10 (ml,p.o) 35.693.25 33.51.82

II.

Toxic control 5 mg/kg 51.782.60 583.266

III.

Piracetam(200mg/kg) 200 (mg, p.o) 20.602.72 10.452.02**

IV. EEBA 200 (mg, p.o) 22.332.02 13.831.887**

V. EEBA 400 (mg, p.o) 41.431.78 26.531.86*

VI. AEBA 200 (mg, p.o) 25.321.97 15.762.39**

VII. AEBA 400 (mg, p.o) 43.333.65 33.822.24*

Values are expressed as mean S.E.M., n=6, significant at *P < 0.05 and **P


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of learning and memory.Finally concluded that, Both the ethanolic and aqueous extracts of the plant possess anti-

diarrhoeal property as indicated by the reduction in the total weight of wet feces, in the experimental models.

Between two extracts ethanolic extract produced most potent anti-diarrhoeal activity. Saponins contribute to the

learning and memory enhancing property. The presence of saponins in ethanolic and aqueous extracts, after

administration of these extracts there was decreased TL on first day as well as second days, indicating significant

improvement of learning and memory. The further research on the plant will be definitely a contributory for

pharmacological establishment.

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