INTRODUCTION:

Liver is one of the largest organs in human body and the chief site for intense metabolism and excretion. So it has a surprising role in the maintenance, performance and regulating homeostasis of the body. It is involved with almost all the biochemical pathways to growth, fight against disease, nutrient supply, energy provision and reproduction (Ward and Daly, 1999). The major functions of the liver are carbohydrate, protein and fat metabolism, detoxification, secretion of bile and storage of vitamin. Thus, to maintain a healthy liver is a crucial factor for overall health and well being.

Liver diseases are mainly caused by toxic chemicals, excess consumption of alcohol, infections and autoimmune disorders. Most of the hepatotoxic chemicals damage liver cell mainly by inducing lipid peroxidation and other oxidative damages (Recknagel, 1983; Wendel et al., 1987; Dianzani et al., 1991).

Drug induced hepatotoxicity has become an important public health care problem contributing to more than 50% of acute liver failure cases. Adverse hepatic events caused by drug can be considered to be either predictable or unpredictable. They may associate with serious morbidity and mortality (Zimmermann, 1999).

Henna is a tall shrub or small tree, 2.6m high. It is glabrous; multi branched with spine tipped branch lets.

Leaves are opposite, entire, glabrous, sub-sessile, elliptical, and broadly lanceolate, acuminate, having depressed veins on the dorsal surface. Henna flowers have four sepals and a 2mm calyx tube with 3mm spread lobes.They is acerbic and used as prophylactic against skin disease. The stewing of leaves is used as mouth wash in sore throat.

The paste of leaves is mostly used for headache and burning sensation in feet. The bark and the seeds of this plant are reported to be used in ayurvedic and unani medicine. The oil obtained from this shrub is used in perfumery purpose. In the present study has been undertaken with the aim to determine the heaptoprotective activity of Lawsonia inermis.

MATERIALS AND METHODS:

Plant Materials:

Dry seeds of Lawsonia inermis were collected from Thanjavur, Tamil Nadu, India. The plant was previously identified and authorized by Dr. P. Prabakaran, Asst.Prof. PRIST University, Thanjavur. The seeds of Lawsonia inermis were dried in shade for 4-6 days. After 6 days, the seeds were made into fine powder using mixer grinder.

Preparation of extract:

The powdered form of Lawsonia inermis was obtained after grinding (100g) was defatted with distilled water using Soxhlet apparatus till exhaustion for about 32 hours (Bose et al., 2007).

Screening of phytochemical compounds:

Chemical tests were carried out on the aqueous extract and on the powdered specimens using standard procedures to identify the constituents as described by Sofowara (1993), Trease and Evans (1978) and Harborne (1973).

Animals:

Albino rats of male sex, approximately 8-10 weeks of age weighing 100-150g were obtained from Animal house, Tamil University, Thanjavur, Tamil Nadu, India. The animals were housed in standard cages, at room temperature (25 ± 3ºC), with 12hr light cycles, all the animals were fed with standard pellet diet. They were given 2 weeks time to get acclimatized with laboratory conditions.

Experimental Design:

The animals were divided into four groups, containing 6 rats in each group. Initial body weight was recorded.

Group I: Animals received standard diet act as control.

Group II: Animal received Paracetamol induction along with normal diet for 7 days

Group III: Animal received aqueous extract (10mg/1ml) of Lawsonia inermis

Group IV: Animal received standard drug Silymarin (10mg/Kg).

Collection of blood:

The blood was collected by Sino-orbital puncture and allowed to clot for few minutes. The clotted blood was transformed to centrifuge tube. The blood was centrifuged at 3000 rpm for 5 minutes. The serum was used for estimation of biochemical parameters such as ALT (Reitman and Frankel, 1957) , AST (Reitman and Frankel, 1957), ALP (Kind,1954), ACP (King et al., 1965) Protein (Reinhold et al., 1953) and Bilirubin (Aniya et al., 2005).

Statistical Analysis:

The results obtained in the present investigation were subject to statistical analysis like Mean and Standard Deviation by Zar (1984).

RESULTS AND DISCUSSION:

Paracetamol is one of the drugs that produce hepatic necrosis and liver damage in over dose has been reported earlier. Alteration in the biochemical parameters reflected marked changes in various enzymatic parameters are Paracetamol treated rats which indicate the most serious toxic effect of Paracetamol.

In this study aqueous extract of Lawsonia inermis were given orally in Paracetamol intoxicated rats by the administration of Paracetamol for 7 days. Biochemical parameters were assessed the protective effect of Lawsonia inermis. The observations were compared with normal rats.

Phytochemical screening:

The preliminary phytochemical investigation of the aqueous fraction showed presence of tannins, phlobatannins, saponins, steroids, flavonoids and terpenoids. According to these results, it may be responsible for the heatoprotective activity. The results were presented in Table.1.

Table 1: Phytochemical screening of Lawsonia inermis

S.No	Test	Result
1	Tannins	+ve
2	Phlobatannins	+ve
3	Saponins	+ve
4	Flavonoids	+ve
5	Steriods	+ve
6	Terpenoids	+ve
7	Cardiac glycosides	-ve

+ indicates Positive; - indicates negative

Levels of liver marker enzymes:

Table-2 indicated the level of marker enzymes in liver. The levels of ALT, AST, ALP and ACP were markedly elevated in Paracetamol induced animals indicating liver damage. Administration of Lawsonia inermis remarkably prevented Paracetamol induced hepatotoxicity and regulate the level of marker enzymes.

Normally, AST and ALP are present in high concentration in liver. Due to hepatocyte necrosis or abnormal membrane permeability, these enzymes are released from the cells and their levels in the blood increases. ALT is a sensitive indicator of acute liver damage and elevation of this enzyme in non hepatic diseases in unusual.ALT is more selectively a liver parenchymal enzyme than AST (Shah,2002).

Assessment of liver function can be made by estimating the activities of serum ALT, AST, ALP and ACP which are enzymes originally present higher concentration in cytoplasm. When there is hepatopathy, these enzymes leak into the blood stream in conformity with the extent of liver damage (Nkosi et al.,2005). The elevated level of these entire marker enzymes observed in the group II, Paracetamol treated rats in this present study corresponded to the extensive liver damage induced by toxin. The reduced concentrations of ALT,AST and ALP as a result of plant extract administration observed during the present study might probably be due in part to the presence of flavonoids.

ACP is frequently employed as a marker enzyme to assess the lysosomal changes invivo because it is localized almost exclusively in the particle and its release parallels that of lysosomal hydrolyses (Tanaka et al., 1968).

Changes in level of Protein and Total Bilirubin:

Table-3 showed the level of Protein and total bilirubin. Increased activity of protein and bilirubin were observed in Paracetamol treated rats. The enhanced activities of Protein and Bilirubin observed in paracetamol fed rats were found to be normalized after administration of Lawsonia inermis.This tendency of these enzymes to return towards near normal level in drug treated rats.

Table 3: The level of Protein and Total bilirubin in normal, induced and treated rats

S.No	Experimental model	Protein (mg/dl)	Total Bilirubin (mg/dl)
1	Control	5.75± 0.02	0.43± 0.01
2	Paracetamol	7.18± 0.04	2.21± 0.05
3	Paracetamol + Lawsonia inermis	6.18± 0.07	0.84± 0.03
4	Paracetamol + Standard drug	5.18± 0.06	0.62 ±0.02

Values are expressed as mean ± standard deviation for 6 animals in each group

Besides, an obvious sign of hepatic injury is the leaking of cellular enzymes into the plasma due to the disturbance caused in the transport functions of hepatocytes (Zimmerman and Seeff, 1970). When liver cell plasma is damaged, a variety of enzymes located normally in cytosol is released into the blood, thereby causing of the cellular proteins in the liver as well hepatic necrosis (Thabrew and Rajatissa, 1987).

CONCLUSION:

From the overall results it can be concluded that aqueous extract of Lawsonia inermis is proved to be one of the herbal remedies for liver ailment.

REFERENCES:

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Received on 24.2.2012 Accepted on 28.04.2012

Asian J. Pharm. Res. 2(2): April-June 2012; Page 75-77

S.No	Experimental Model	ALT (mg/dl)	AST (mg/dl)	ALP (mg/dl)	ACP (mg/dl)
1	Control	51.21± 0.3	60.32± 0.12	30.09± 0.14	4.02± 0.01
2	Paracetamol	200.21± 0.4	180.21±0.16	90.62 ±0.51	9.32 ±0.21
3	Paracetamol + Lawsonia inermis	57.02± 0.6	71.31± 0.18	39.21± 0.42	4.81± 0.32
4	Paracetamol + Standard drug	50.21 ±0.7	63.17± 0.12	32.33± 0.54	4.56± 0.25