V. I. Hukkeri, Kusum S. Akki*, R. R. Sureban, B. Gopalakrishna, V. V. Byahatti and S. V. Rajendra

Department of Pharmacognosy and Phytochemistry, K. L. E. S’s College of Pharmacy, Vidyanagar, Hubli–580 031, India.

*Corresponding Author:
Kusum S. Akki
Department of Pharmacognosy and Phytochemistry, K. L. E. S’s College of Pharmacy, Vidyanagar, Hubli–580 031, India
E-mail: kusumakki@yahoomail.com
Date of Submission 09 June 2005
Date of Revision 21 October 2005
Date of Acceptance 14 August 2006
Indian J Pharm Sci,2006, 68 (4): 542-543  

DOI: 10.4103/0250-474X.27844

Abstract

The decoction of the leaves of Nyctanthes arbor-tristis Linn. of Oleaceae widely used in Ayurvedic system of medicine for the treatment of sciatica, arthritis, fevers, various painful conditions and diuretics, liver disorders and as laxative. The aim of the present study was to evaluate the alcoholic and aqueous extracts of the leaves of Nyctanthes arbor-tristis for hepatoprotective effect against carbontetrachloride-induced liver damage in rats.Administration of alcoholic and aqueous extracts of the leaves of Nyctanthes arbor-tristis protect the liver from toxic effects of carbontetrachloride by reducing the elevated levels of Serum glutamate pyruvate transaminase, Serum glutamate oxaloacetate transaminase and serum bilirubin (total and direct). Results revealed that both the alcoholic and aqueous extracts showed significant hepatoprotective activity by reducing the elevated levels of biochemical parameters at a dose of 500 mg/kg body weight. The results were supported by histopathological studies of liver samples which showed regeneration of hepatocytes by the extracts.

Liver plays a major role in detoxification and excretion of many endogenous and exogenous compounds, any injury to it or impairment to its functions may lead to many implications on one’s health. Management of liver disease is still a challenge to the modern medicine [1,2]. Modern medicine has little to offer alleviation of hepatic ailments; where as most important representatives are of phytoconstituents. Nyctanthes arbor-tristis Linn. is a large shrub which is widely cultivated throughout India as a garden plant. The bitter leaves are used in traditional system of medicine for the treatment of rheumatism, sciatica and intestinal worms. The powdered seeds are recommended for the treatment of scurvy [3-5]. In this study an attempt was made to provide scientific backing to the traditional claims.

The leaves of Nyctanthes arbor-tristis were collected from the local areas of Hubli and Belgaum in the month of April and the same was authenticated at Department. of Botany, Karnataka University, Dharwad. The dried powdered leaves were exhaustively extracted with 95% ethanol in a Soxhlet apparatus and also macerated with chloroform water for 7 days. The extracts were further concentrated in vacuum under pressure using rotary flash evaporator and dried in descicator.

Both the extracts were suspended in water using 1% Twen-80 and subjected for hepatoprotective activity in CCl4-induced hepatotoxicity. Acute toxicity studies were conducted by using albino mice of either sex weighing between 20 and 25 g and of 90 days age. The animals were fasted over night prior to the experimental procedure. The method of Up and Down or ‘Staircase’ was used to determine the dose [6], Tween-80 (1%) was used as a vehicle to suspend the extracts. Wistar rats (150-200 g) of either sex were used for hepatoprotective activity. The animals were grouped into five groups of six animals each and maintained on standard diet and water, ad libitum. All the animal experimental protocol has been approved by the Institutional Animal Ethics Committee.

Hepatoprotective activity [7] was carried out using Wistar rats (150-200 g) of either sex. The animals were divided in to five groups of six animals each and maintained on standard diet and water, ad libitum. Tween-80 (1%) was given to groups 1 and 2 as a vehicle for 10 days by oral route. Liv- 52 was administered to group 3 at the dose of 1 ml per kg body weight by oral route for 10 days. Ethanol and aqueous extracts were administered to groups 4 and 5, respectively at a dose of 500 mg/kg by oral route for 10 days. CCl4 at a dose of 0.7 ml per kg body weight was injected to animals of groups 2, 3, 4 and 5 on 3rd, 6th and 10th day by intraperitonial route. On 10th day, 1 h after the last dose of Carbon tetrachloride injection, animals were sacrificed by cervical dislocation and the blood was collected from the carotid artery, serum was separated and used for the estimation of various biochemical parameters.

Biochemical parameters [8] such as glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, alkaline phosphatase (ALP) and serum bilirubin (total and direct) were determined (Table 1). Liver was excised quickly fixed in 10% formalin and then fixed in bovine solution, they were processed for paraffin embedding following the standard micro technique. Sections of liver were stained with haematoxylin-eosin and were observed microscopically for any histopathological changes.

Group Dosemg/kg SGPT(IU/L) SGOT(IU/L) ALP(IU/L) Bilirubin mg/dl
Total Direct
Control - 158.2±3.499 227.4±7.054 12.32±1.930 0.74±0.04 0.22±0.02
CCl4 0.7 273.2±3.455* 297.4±16.567* 15.22±0.205 1.4±0.07* 0.22±0.02
Standard 1.0 160.8±2.131** 226.6±2.441** 12.72±0.159 0.78±0.02** 0.26±0.02
Alcoholicextract 500 161.4±3.076** 229.4±3.541** 12.94±0.139 0.84±0.02** 0.22±0.02
Aqueousextract 500 160.2±1.844** 236.2±3.736** 12.88±0.164 0.84±0.02** 0.22±0.02

Table 1: Effect of nyctanthesarbor-Tristislinn. On ccl4-Induced hepatotoxicity in rats

The mean value±SEM was calculated for each parameter, each parameter was analyzed separately using ANOVA followed by Dunnets ‘t’ test.It is revealed that the alcoholic and aqueous extract contains flavonoids, sterols, tannins, carbohydrates and glycosides (flavonoidal). Results revealed that both the alcoholic and aqueous extract of leaves of Nyctanthes arbor-tristis Linn exhibited an ability to counter act the CCl4-induced hepatotoxicity by decreasing the elevated enzyme levels in the blood compared to the CCl4 group (P<0.01).

Histopathology of liver from normal control group shows prominent central vein, normal arrangement of hepatic cells. Microscopical examination of carbon tetrachloride treated liver section shows various degrees of pathological changes starting from centrilobular necrosis of hepatic cells and central lobular fatty regeneration. Liver section of standard treated and from Nyctanthes arbor-tristis treated groups shows moderate protection in CCl4-induced liver damage.

Since the results of hepatoprotective activity showed a significant decrease in the elevated levels of serum enzymes and histopathological results showed a significant regeneration of hepatocytes. Thus, from the studies we may conclude that the ethanolic and aqueous extracts of the leaves of Nyctanthes arbor-tristis Linn. can be used as hepatoprotective. The results were also comparable with standard drug.

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