Antiurolithiatic activity of aqueous extract of bark of moringa oleifera (lam.) in rats

doi:10.4236/health.2010.24053

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Vol.2, No.4, 352-355 (2010) Health

doi:10.4236/health.2010.24053

Antiurolithiatic activity of aqueous extract of bark of

moringa oleifera (lam.) in rats

Jameel Fahad1, Vijayalakshmi1, M. C. Satish Kumar1, Sanjeeva1, G. Prabhath Kodancha1,

Benegal Adarsh1, A. L. Udupa2*, U. P. Rathnakar3

1Department of pharmacology, Kasturba Medical College, Manipal, India

2Departmen t of Pharmac ology, Faculty of Medical Sc ience s, Univer sity of the West Indie s, Cave Hill, Ba rba dos;

aludupa2002@yahoo.com

3Department of pharmacology, Kasturba Medical College, Mangalore, Karnataka, India

Received 8 January 2010; revised 20 February 2010; accepted 22 February 2010.

ABSTRACT

In the present study, aqueous extract of bark of

Moringa oleifera administered orally, was eva-

luated for its antiurolithiatic potential in albino

rats of Wistar strains. The stones were pro-

duced in this study by zinc disc foreign body

insertion in the bladder supplemented with 1%

ethylene glycol in drinking water. The reduction

in weight of the stones was used as criteria for

assessing the preventive or curative antiuro-

lithiatic effect of the bark of this plant. Two

doses of extract for prophylactic and curative

groups were used. In both groups the oral ad-

ministration of the extract of bark of Moringa

oleifera has resulted in significant reduction in

the weight of bladder stones compared to the

control group.

Keywords: Moringa oleifera bark; Antiurolithiatic

activity; Introduction

1. INTRODUCTION

Moringa oleifera is an important medicinal herb, be-

longing to the family Moringaceae. It is considered as

miracle tree as all the parts of the plant are useful for

human health. It is also known as drum stick tree,

horseradish tree, clarifier tree and mother’s best friend

in different parts of the world, based on their appear-

ance and unique uses. It is mostly cultivated all over

the plains of India, in the old world tropics and it is

used medicinally in Guinea, Madagascar, and Burma

[1-3].

The various parts of the plant used include flower,

pod, leaves, bark and roots. The flowers, after cooking,

are eaten either mixed with other foods or fried in

butter and have been shown to be rich in po tassium and

calcium [4]. The leaves are eaten as greens, in salads,

in vegetable curries, as pickles and for seasoning. The

Bark is regarded as an antiscorbic, and it exudes a

reddish gum sometimes used for the treatment of

diarrhoea. The ro ot s are bitter, act as a tonic to the body

and lungs, and have an expectorant activity. They are a

mildly diuretic and are used as a stimulant in paralytic

afflictions, epilepsy and hysteria. The pods are the most

valued and widely used of all the tree parts. The pods

are extremely nutritious, containing all the essential

amino acids along with many vitamins and other nu-

trients. The immature pod can be eaten raw or prepared

like green peas or green beans, while the mature pods

are usually fried and possess a peanut- like flavour. The

pods also yield 38-40% of non-drying, edible oil

known as Ben Oil which is clear, sweet and odourless,

and never becomes rancid. Overall, its nutritional value

most closely resembles olive oil [4].

In recent decades, many scientific studies using the

extracts of leaves, pods and roots of Moringa oleifera

are being carried out to confirm many potential uses

including wound healing [5,6], anti-tumour [7], antihepa-

totoxic [8], antifertility [9], hypotensive [10], antiuroli-

thiatic [11] acute anti-inflammatory [6,12,13], and anal-

gesic activity [14].

Even though the root extracts of Moringa oleifera

have been studied for diuretic activity [11], there is no

scientific study showing antiurolithiatic activity of Mor-

inga oleifera bark extract. Therefore this study was

conducted in order to evaluate its antiurolithiatic activ ity

in rats.

2. MATERIALS AND METHODS

2.1. Approval for the Project

Approval for the experiment was obtained from the

Institutional animal ethics committee (IAEC), Kasturba

Medical College, Manipal, vide letter No. IAEC/KMC

07/2008-2009.

F. Jameel et al. / Health 2 (2010) 352-355

353

2.2. Plant Material

The bark of Moringa oleifera Lam. available locally

were collected be tween February and May in and aroun d

Udupi. The botanical identity has been confirmed by

department of botany, Sri Poornapagna College, Udupi.

The voucher specimen is preserved in our laboratory for

future reference.

2.3. Method of Extraction

The fresh bark collected were cut into small pieces, par-

tially crushed and soaked in water overnight which was

later subjected to boiling for 6 hours. The resultant ex-

tract was then drained and concentrated in a water bath

to get the concentrated extract. The yield of the extract

was 10.5%. The extract was stored in desiccators and

used for further experiments after dissolving it in dis-

tilled water.

2.4. Animals

Inbred albino rats of Wistar strain, of either sex, aged

around 2 to 3 month s and w eigh ing 15 0-2 00 g w er e used.

They were housed in standard conditions of temperature

(25 ± 2°C), relative humidity of 45-55%, and maintained

on 12–hour light: 12– hour dark cycle in animal house of

Kasturba Medical College, Manipal. They were fed

standard pellet diet (Hindustan Lever rat pellets) and

water ad libitum.

3. ACUTE TOXICITY STUDY

The aqueous extract of Moringa oleifera bark was ad-

ministered orally in the escalating dosages, up to 8 g/kg

to different groups of rats (n = 6, in each). The animals

were observed for behavioural and physiological varia-

tions initially continuously for 4 hours, followed by 4th

hourly for 12 hours and there after once daily for 14

days.

3.1. Antiurolithiatic Study: Method of

Induction of Urolithiasis by Insertion of

Zinc Disc [15-19]

Rats were anaesthetized with intraperitoneal ketamine

(50 mg/kg). A suprapubic incision was made and the

abdomen was opened. The urinary bladder was then

carefully exposed and the urine in the bladder was as-

pirated with a sterile syringe. A small nick was made at

the apex end of urin ary bladder and the ster ile zinc disc

(previously weighed) was carefully inserted into the

bladder. Then the bladder was closed in a single stitch

using chromic catgut (4-0). The abdomen was then

closed in layers with chromic catgut and skin was

closed with silk thread. The rats were allowed to re-

cover from anaesthesia. Food and 1% ethylene glycol

in water was given ad libitum. The stone was allowed

to form and grow inside the bladder during the study

period. After the study period the rats were sacrificed

and zinc disc implants/stones were removed from the

bladder and dried. Stones taken out were weighed. The

difference between initial and final weights indicated

the amount of stone formed.

3.2. Grouping of Animals for Different

Treatments and Procedure of the Study

Adult albino rats of Wistar strain, weighing between

150-200 g, were selected for the study. In this study us-

ing aqueous extract of bark of Moringa oleifera, the ra ts

were divided into 6 grou ps with 6 animals in each group

receiving different treatments. Group I–Prophylactic

control (1% ethylene glycol for 4 weeks), Group II–Pro-

phylactic treatment (1% ethylene glycol+aqueous extract

of bark of Moringa oleifera 400 mg/kg orally for 4

weeks), Group III–Prophylactic treatment (1% ethylene

glycol+aqueous extract of bark of Moringa oleifera 8 00 mg/

kg orally for 4 weeks), Group IV–Curative control (1%

ethylene glycol for 4 weeks followed by water for 4

weeks), Group V–Curative treatment (1% ethylene gly-

col for 4 weeks followed by aqueous extract of bark of

Moringa oleifera 400 mg/kg for 4 weeks), Group VI–

Curative treatment (1% ethylene glycol for 4 weeks

followed by aqueous extract bark of Moringa oleifera

800 mg/kg for 4 weeks).

Prophylactic activity against urolithiasis was tested

using Groups I to III in this study and after 4 weeks,

animals were sacrificed and vesical calculi were col-

lected, weighed and statistically evaluated.

Curative property was tested in using Groups IV to VI

in the stud y and, at the end of eig ht weeks , animals were

sacrificed and vesical calculi were collected, weighed

and statistically evaluated.

3.3. Weight of Stones

The difference between the weight of the implanted zinc

discs at the time of implantation and final weight of the

dried calculi taken out from the bladder at the end of the

4th and 8th week period indicated the weight of deposited

stone.

4. STATISTICAL ANALYSIS

The data obtained from the study was statistically evalu-

ated using a parametric test ANOVA (Analysis of Vari-

ance) and Tukey as post hoc test. This was done with

SPSS (Statistical package for social science) software.

5. RESULTS

The results of different groups studied using aqueous

F. Jameel et al. / Health 2 (2010) 352-355

354

Table 1. Weight of stone material deposit on zinc discs in con-

trol and after using aqueous extract of bark of Moringa oleifera

400 mg/kg & 800 mg/kg treated rats.

Groups No.of rats

Weight of stone (mg)

(Mean ± SEM)

Group I–prophylactic control 6 278.16 ± 7.35

Group II–prophylactic treat-

ment (400 mg/kg) 6 139.5 ± 8.89*

Group III–prophylactic treat-

ment (800 mg/kg) 6 105.6 ± 5.61*

Group IV–curative control 6 269 ± 10.45

Group V–curative treatment

(400 mg/kg) 6 170.6 ± 12.98*

Group VI–curative treatment

(800 mg/kg) 6 114.6 ± 4.43*

* p < 0.001, df = 2, SEM = Standard error of mean, n = 6

extract of bark of Moringa oleifera 400 mg/kg & 800 mg/

kg are shown in Table 1. In this study, the extracts from

bark of the plant showed significant decrease in the

weight of stones compared to control after the study pe-

riod.

6. DISCUSSIONS

In the present study, aqueous extract of bark of Mor-

inga oleifera 400 mg/kg & 800 mg/ were evaluated

for the antiurolithiatic potential in albino rats of Wis-

tar strains. The method used for induction of stones in

this study was zinc disc foreign body insertion tech-

nique supplemented with 1% ethylene glycol, which

is commonly used in these type of studies.

In this study, the weight of the stones was used as

criteria for assessing the preventive or curative effect

of the bark of Moringa oleifera. Two doses of extract

for prophylactic and curative groups were used. In

both groups the administration of the extract have

resulted in significant (p < 0.01) reduction in the weight

of stones compared to the control group.

The decrease in the bladder stone formation was in-

consistent with the increase in the dose of the extract

which could be due to variability in the response due to

physiological variation. The stone formation in the con-

trol group itself was variable to a certain extent ranging

from 240 mg to 300 mg in prophylactic control group

and 248 mg to 310 mg in curative control group, which

is indicative of normal physiological variation. Among

the different strains of rats used for preclinical studies,

Wistar rats are much less susceptible to persistent blad-

der infection and struvite stone formation in comparison

to other strains. In a study where female Fischer 344

(F344), Lewis (LEW), Sprague-Dawley (SD), and Wistar

(WIS) rats were inoculated with a host-adapted strain of

Ureaplasma parvum, and necropsied at 2 weeks post

inoculation; 100% of F344, 42% of SD, 10% of LEW,

and 10% of WIS rats remained infected. Severe bladder

lesions and struvite calculi were seen in 64% of F344

rats; in other rat strains, bladder lesions were mild or

absent [20].

7. CONCLUSIONS

In conclusion we can confiden tly confirm the possibility

of antiurolithiatic activ ity of bark of Moringa oleifera as

there was reduction in size of the stones. Further studies

are needed to prove the stone dissolving property of

aqueous extract of bark of Moringa oleifera (400 mg/kg

& 800 mg/kg) in other animal models

8. ACKNOWLEDGEMENT

We the authors are thankful to the Dean of Kasturba Medical College,

Manipal and to the K.M.C. Trust Manipal for the financial support

rendered for this study.

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