Effect of Allium sativum in Immoblilization Stress Induced Albino Rats.

 

Roshan S.2, Abdullah Khan2, Sadath Ali1* and Tazneem B.1

1Luqman College of Pharmacy, Gulbarga, Karnataka.

2Department of Pharmacy, Singhania University, Rajasthan.

ABSTRACT:

The present study was carried out to evaluate the effect of ethanolic extract of Allium sativum (Alliceae) on immobilization stress induced albino rats. The effect was assessed by estimation of various  biochemical parameters like glucose, cholesterol, triglycerides and blood urea nitrogen (BUN), and by determining the  weight of organ such as, liver, spleen, testes, adrenal gland, blood cell count (WBC)  and also the differential count at a dose of 200mg/kg body weight per oral. The extract showed significant (p<0.001) decrease in blood glucose, cholesterol, triglyceride and BUN and also decreased the weight of organs. It also showed a significant (p<0.05) decrease in weight of adrenal gland.  A significant (p<0.01) decrease in WBC count, polymorphs and monocytes and decrease in lymphocytes (p<0.05) and eosinophils was observed, compared to control group. Thus the obtained results revealed that the Allium sativum has got a significant anti stress activity.

 

KEYWORDS: Immobilization stress, Allium sativum, Cortisol, WBC, BUN.

 

INTRODUCTION

Stress is the body’s physical, mental or chemical reaction observed usually during excitation, confusion or in unsafe condition. A large proportion of all illness is believed to occur due to stress closely associated with modernization of life. Fortunately Mother Nature has an answer to this challenge - a unique class of herbal products called "adaptogens". Adaptogens have the broad-spectrum healing properties of any herbal medicines, but their unique value is that they specifically relieve stress. According to modern science adaptogens are natural plant products that increase the body's ability to cope with internal and external stress factors, and normalize the functions of the organism1. Stress is particular response of the body to external event or stress full situation. Body responds by releasing certain hormones like adrenaline and cortisol1. This leads to an increase in heart rate, blood pressure and metabolic rate, and also causes diabetes, depression, and anxiety2.

 

The bulb Allium sativum belongs to family Alliaceae, perineal bulbous scapigerous herb. It is cultivated throughout India. It has great medicinal value, bulb used has anti diabetic, anti-cancer, anti hypertensive, and lipidemic activity. The juice is effective in rheumatism, whooping cough, tuberculosis and has anti bacterial and rejuvenative actions. The fresh bulb is used as rejuvenative. The plant contains proteins, amino acids, flavonoids, hydrocarbons, allin, sulphur amino acid, anthocyanins, kaempferal and carbohydrates3. The plant reported to have anti diabetic4, anti-oxidant5, hepataprotective6, anti stress7 and anti lipidemic8 effects. In the present study ethanolic extract of Allium sativum was evaluated for its effects on adaptogenic activity.

MATERIALS AND METHOD:

The plant Allium Sativum was collected from local area of Gulbarga, and Authentificaton was done by Dr. Seetharam. G,   Department of Botany, Gulbarga University, Gulbarga. A voucher specimen number LCP- 216. The same has been deposited in research laboratory. Fresh bulb crushed and pulverized in laboratory, extracted with 95% ethanol in percolator. The extract was concentrated by using rotary flash evaporator. The residue was dried in desicator over sodium sulphate.

 

Experimental animals:

Adult Swiss albino mice (20- 25g) and Wistar rats (150 -200g) of either sex were used for the study. The mice and rats were fed with standard pellet (Parnava Agro industries Ltd. Sangali, India) and water ad libitum. The animals were maintained under standard 12-hr light / dark cycle throughout the study. The study protocol was approved by IAEC. (No.CPCSEA/IAEC/PC-01/346)

 

Acute toxicity study:9

The study was performed according to the acute toxic classic method (as per CPCSEA/OECD guidelines). Swiss albino mice were used for acute toxicity study. The animals were kept Fasting for overnight providing only water, after which the test drug extract dissolved in Water was administered orally at the dose of 800 mg/kg and observed for 14 days. Animals were observed individually at least once during the first 30 min after dosing, periodically during the first 24 h (with special attention during the first 4 h) and daily thereafter for a period of 14 days. Once daily cage side observations included changes in skin and fur, eyes and mucous membrane (nasal) and also respiratory rate, circulatory(heart rate and blood pressure), autonomic (salivation, lacrimation, perspiration, piloerection urinary incontinence, and defecation) and central nervous system (ptosis, drowsiness, gait, tremors and convulsion). The toxicity study carried out as per the guidelines of AOT- 421 using albino mice. The extracts were found to be safe till 400mg/kg. Hence we selected 200mg/kg dose for pharmacological screening.

 

Immobilization stress:

Albino rats (210-230gm) of either sex were divided into 4 groups of 6 animals each. Group -1 served as control, Group-2 served as immobilization stress control , Group-3 served as Allium sativum (200mg/kg) treatment group and Group-4 served as panax ginsang(50mg/kg) treatment. Immobilization stress was induced in 2nd , 3rd and 4th  group animals by immobilizing the animals with head down in supine position, by fixing the animals to a board inclined position at an angle of 60°  daily for 2hrs for the duration of 10 days10. On 10th day, the animals were sacrificed and blood was collected for estimation of biochemical parameter like glucose11, cholesterol12,, triglyceride  , BUN 13, blood cell count14, and weight of organs like liver , spleen, testes  and adrenal gland15.

 

Statistical analysis:

Analysis of data was performed using SPSS V 11.0 statistical software. Descriptive statistics were adopted to display data in Mean ± SEM.  One way ANOVA was used to compare the mean value obtained between the different groups (Specify the test performed for post t test).  Differences were considered significant (p<0.05).

 

RESULTS:

The ethanol extract of Allium sativum has significantly (p<0.001) reduced the elevated levels of biochemical parameters  glucose, cholesterol, BUN and triglyceride with compared to stress control group results are shown in Table 1.  The extract has also reduced all the blood cell count WBC’s significantly (p<0.01), except lymphocytes and Eosinophils (p<0.05) compared to stress control group Table 2. Determination of weight of organs showed that the extract has significantly (p<0.01) reduced the weight of liver, spleen and testes, however it showed no effect on weight of adrenal gland as shown in Table 3.

 

DISCUSSION:

Under stressful conditions cortisol and corticosterone will be secreted by adrenal cortex18 Hyper secretion of cortisol helps in the maintenance of internal homeostasis through the process of gluconeogenesis and lipogenesis19. Treatment Allium sativum extracts significantly reduced the hyperglycemia by reducing the hyper activity of adrenal cortex and also by maintenance homeostatic mechanism. This was further in agreement with plasma corticosterone levels. The mechanism by which stress raises serum cholesterol is likely to be related to the enhanced activity of hypothalamo-hypophyseal axis resulting in increased liberation of catecholamines and corticosteroids20. In the present study a significant increase in plasma corticosterone was observed in cold stress. After treatment with Allium sativum it was reduced in cholesterol level might be increased due to the mobility of fat which could not be suppressed by extracts. The effect of stress on serum triglycerides has been shown to be variable probably catecholamines mobilize lipids from adipose tissues. Cold stress models showed an increase in triglyceride levels. Allium sativum extracts were not able to suppress the stress induced increase in triglycerides levels. BUN levels were increased in cold stress as these are the end products of protein metabolism. In excess adrenocortical activity21, due to increased metabolism of protein increases urea excretion, in the present study a similar effect was observed. Extracts of Allium sativum decreased the BUN levels as compare to stress control, indicating a diminished catabolism of protein under stressful conditions. Adrenal glands and liver weights were significantly increased in and cold stress than control group. Stress induces adrenomedullary response in man to release adrenaline which in turn stimulates β2 receptors on the pituitary gland. It leads to greater release of ACTH that can stimulate the adrenal medulla as well as cortex19.

 

 

Table 1  Effect of Allium sativum on Biochemical Parameter in immobilization stress induced albino rats.

Parameter

Normal

Immobilization  stress

Allium Sativum extract.

Panax ginseng extract.

Glucose

80.24  ±  0.812

118.2 ± 2.41

98.25 ± 5.21***

87.15 ± 4.91

Cholesterol

40.21 ± 1.882

62.4 ± 1.98

49.69 ± 2.03**

44.79 ±  1.93

Triglyceride

71.24 ± 0.712

110.24 ± 2.94

88.94 ± 1.76**

83.84 ± 1.46

BUN

30.14 ± 0.512

51.41 ± 2.61

43.98 ± 1.22**

43.78 ± 1.42

 

Table 2  Effect of Allium sativum on weight of organ in immobilization stress induced albino rats.

Parameter

Normal

Immobilization stress

Allium sativum extract.

Panax ginseng extract.

Spleen

3.57  ± 0.11

2.891 ± 0.12

3.08 ± 0.29**

3.11 ± 0..21

Testes

1.681 ± 0.06

1.421 ± 0.02

1.341 ± 0.29**

1.421 ± 0..03

Liver

5.142 ± 62.5

6.818 ± 38.2

5.921 ± 13.8*

5.461 ± 19.76

Adrenal gland

0.250 ± 0.01

0.423  ±0.44

0.358 ± 0.01*

0.398 ± 0.22

 

Table 3 Effect of Allium sativum on blood cell count in immobilization stress induced albino rats.

Parameter

Normal

Immobilization stress

Allium Sativum extract.

Panax ginseng extract.

WBC’s count

4824 ± 41.02

7540 ± 166.7

6124 ± 44.2***

5817 ± 48.21

Lymphocytes

48 ± 1.72

71 ± 1.29

64 ± 0.72**

59 ± 1.03

Monocytes

1.00 ± 0.0

3.25 ± 0.19

1.75 ± 0.50**

1.5 ± 0.76

Neutrophills

24 ± 0.40

35 ± 0.84

28 ± 0.50*

31 ± 0.22

Eosinophils

1.25 ± 0.4

4.50 ± 0.25

2.25 ± 0.25*

--

 

 

Resulting in further release of adrenaline and increase in weight of adrenal gland to greater extent. Cortisol increases m-RNA levels in liver cells, increase in liver weight and it was well in correlation with the observed .increase in plasma corticosterone levels. Spleen contracts during stress and releases more amount of blood (RBC) into circulation hence its weight decreases. Stress induced increase in weight of liver and adrenal glands and a decrease in spleen weight indicating their protective effect against stress15. Stress causes alteration in hematological parameters like increase in WBC and DLC counts, neutrophils were increased more significantly in cold stress stress. Extract reduced the WBC, lymphocytes, eosinophils and monocyte counts in cold stress20.

 

The adaptogenic effect of Allium sativum so observed is attributed to the presence of Flavonids, phenolic glycosides, hydrocarbons, proteins, and vitamins. The results of study suggest that the bulbs of Allium sativum have got significant adaptogenic activity. we are studying on other adaptogenic models.

 

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Received on 13.05.2010

Accepted on 31.05.2010     

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Research J. Pharmacology and Pharmacodynamics. 2(4): July-August 2010, 293-295