INTRODUCTION:

The liver disorders are one of the world problems. Despite its frequent occurrence, high morbidity and high mortality, its medical management is currently inadequate. So far not yet any therapy has successfully prevented the progression of hepatic disease, even though newly developed drugs have been used to treat chronic liver disorders, these drugs have often side effects^[1].

Liver inj-ury due to chemicals and infectious agents may lead to progressive liver fibrosis and ultimately cirrhosis and liver failure^[2].

Carbon tetrachloride is widely used for experimental induction of liver damage. The principle causes of carbon tetrachloride is induced hepatic damage in lipid peroxidation and decreased activities of antioxidant enzymes and generation of free radicals. Various medicinal plants have been used to treat for various diseases all over the world. Nowadays, Indian medicinal plants are belonging to about 40 families were investigated as liver protective drugs^[6,11].

MATERIALS AND METHODS:

Plant material:

Whole plant of Leucas cephalotes were collected from kailsa, U.P. and authenticated from botany department, IFTM University Moradabad.

Animals:

Wistar Albino rats of either sex weighing between 150-200 gm aged 12 weeks were employed for this investigation. They were kept under temperature of 23°C, humidity of 50 % and 10 h : 14 h of light and dark cycles, respectively. They were housed individually in polypropylene cages comtaining sterile paddy husk. Before and during the experiment, rats were fed wiyh standard diet. Animals were fasting overnight and weighed before the experiment.

Drugs and chemical reagents:

Paracetamol

Silymarin

Study Design and Experimental Protocol:

The rats were randomly divided into seven groups of six rats. Rats in Group I (Control) received 2mL of 2% gum acacia p.o. for 8 days. Group II were treated by 2% gum acacia + PCM (2 g/kg) p.o., Group III by silymarin (25 mg/kg) i.p, while Group IV, V and VI received ethanolic plant extract p.o. at 100, 200 and 400mg/kg respectively. Plant extract, Paracetamol were administered orally by suspending in 2mL of 2% gum acacia. The standard hepatoprotective drugs, silymarin were given orally and intraperitoneally, respectively. All reagents were administered daily for 7 consecutive days. And on the 8th day single dose of Paracetamol 2g/kg was given to all group rats except rats in group I.

On the 9th day, after 24 hours of Paracetamol administration blood samples were collected by direct cardiac puncture under light ether anesthesia. Serum was separated by centrifuging at 2500 rpm for 15 min and used for analysis of various biochemical parameters. All rats were sacrificed by cervical dislocation, and livers were removed for histopathological examination^[9].

Statistical analysis:

All procedure were carried out in triplicate and the results expressed as standard error of mean (SEM). The data were analysed by one way ANOVA using GraphPad Prism software (Version 5.01)

Table 1- Biochemical Parameters Evaluation

Treatment	Dose	SGOT	SGPT	Total Protein	Serum Albumin
Normal control	2ml of 2% gum acacia p.o.	55.265±0.467	72.280±0.528	6.290±0.072	4.160±0.022
PCM Group	2% gum acacia + 2g/kg PCM p.o.	169.345±3.522	215.410±1.319	2.540±0.174	1.405±0.007
Silymarin	25mg/kg i.p.	80.125±0.686****	77.440±0.438****	6.410±0.112****	3.365±0.069****
Ethanolic Extract	100mg/kg p.o.	102.925±0.208****	193.725±3.459****	2.735±0.078	1.350±0.018
Ethanolic Extract	200mg/kg p.o.	111.195±0.566****	174.425±1.438****	3.060±0.022	2.780±0.040****
Ethanolic Extract	400mg/kg p.o.	83.930±1.172****	85.350±1.655****	5.795±0.078****	3.535±0.168****

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Table 2- Biochemical Parameters Evaluation

Treatment	Dose	Globulin	Total Bilirubin	ALP
Normal control	2ml of 2% gum acacia p.o.	2.495±0.029	0.485±.011	95.970±0.018
PCM Group	2% gum acacia + 2g/kg PCM p.o.	1.430±0.098	1.990±0.009	184.695±1.447
Silymarin	25mg/kg i.p.	2.170±0.009****	0.940±0.013****	106.510±1.319****
Ethanolic Extract	100mg/kg p.o.	1.525±0.020	1.693±0.148	173.445±0.901****
Ethanolic Extract	200mg/kg p.o.	1.685±0.123	1.647±0.159	160.335±2.202****
Ethanolic Extract	400mg/kg p.o.	2.000±0.004****	0.850±0.018****	109.195±0.119****

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001

Histopathological examination:


A: Normal histology and central vein were appeared and not seen in slide	B: Inflammation in the liver cell was seen	C: Less cytoplasmic degeneration was occurred


D: Dialated sinusoids	E: Degeneration was occurred	F: Less cytoplasmic degeneration was occurred

Fig. 1- Histopathology of liver of wistar rat treated with Ethanolic extract.

Fig. 2 SGOT value in treatment group Fig. 3- SGPT value in treatment group

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM,n=6, P**<0.01, ***P<0.001, P****<0.0001.

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Fig. 4- Total protein value in treatment group Fig. 5: Serum albumin value in treatment group

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Fig. 6 – Globulin value in treatment group Fig. 7- Total Bilirubin value in treatment group

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as

Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001.

Fig . 8 – ALP value in treatment group

Data was analysed using one way ANOVA followed by Dunnett Test. Value are expressed as Mean±SEM, n=6, P**<0.01, ***P<0.001, P****<0.0001

RESULTS AND DISCUSSION:

Results:

Paracetamol treated rats showed significant increase in the SGOT, SGPT, serum albumin, bilirubin, ALP and decrease in total protein, globulin. While ethanolic extract of Leucas cephalotes showed significant hepatoprotective activity. Paracetamol causes necrosis and haemorrhages in hepatic tissues. The mean weight of the liver was decreased in Paracetamol treated group while doses of Ethanolic Extract increased mean weight of the rats. These increases were comparable to standard Silymarin.

DISCUSSION:

Paracetamol is a known antipyretic and analgesic which produces hepatic necrosis in high doses. Results of our study showed that Paracetamol causes increased levels of SGOT, SGPT, serum albumin, bilirubin and ALP and decrease in total protein, globulin. After administration of the plant extract, Silymarin, the levels of the enzymes were restored. The drugs like Silymarin and the plant extract decreased increase enzyme levels in tested groups, indicating the protection of liver cell membrane or regeneration of damaged liver cells. The Ethanolic Extract of Leucas cephalotes as shown the ability to maintain the normal functioning of the liver. From the above study, we conclude that the Ethanolic Extract of Leucas cephalotes is proved to be one of the herbal remedies for liver disease especially for Paracetamol induced hepatotoxicity.

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Received on 18.07.2019 Modified on 15.09.2019

Research J. Pharm. and Tech 2020; 13(3):1183-1186.

DOI: 10.5958/0974-360X.2020.00218.8