Tylophora asthmatica L. Prevents Lipid Peroxidation in Acetaminophen Induced Hepato Toxicity in Rats

 

R. Malathi1 S. Ahamed John2 and A.Cholarajan3

1Department of Biotechnology, Bharathidasan University College (w), Orathanadu, Thanjavur, Tamilnadu, India.

2Department of Botany, Jamal Mohamed College, Trichy, Tamilnadu, India.

3Department of Microbiology, Meenakshi Chandrasekaran College of Arts and Science, Pattukkottai – 614 626, Thanjavur, Tamilnadu, India.

*Corresponding Author E-mail: prashardeepak99@yahoo.in

 

ABSTRACT:

Methanolic extract of Tylophora asthmatica leaves were prepared and tested for its antilipidperoxidation effect against acetaminophen induced hepato toxicity in rats. Alteration in the levels of lipid peroxide, iron, ferritin, total cholesterol, total lipid and phospholipids were tested in both experiment and control group of rats. Acetaminophen [1gm/kg] has showed a significant increase [p<0.05] in experimental rats except the level of phospholipids, which showed a significant decrease [p<0.05] compared to control rats. Treatment with methanolic extract of Tylophora asthmatica leaves [200mg/kg] has brought back the altered levels of this parameter to the near normal levels in the dose dependent manner.

 

KEYWORDS: Acetaminophen, Hepatoprotective, Tylophora asthmatica.

 

INTRODUCTION:

The liver is a vital organ of paramount importance involved in the maintenance of metabolic functions and detoxification from the exogenous and endogenous chronic alcoholism. If during all such exposures to the above mentioned challenges the natural protective mechanisms of the liver are overpowered, the result is hepatic injury.

 

Acetaminophen (or) paracetamol overdose is a specific hepatotoxic agent, its effect being confined to the liver previous reports shown that excess level of the ROS formation by NAPQF leads to enhancement of lipid peroxidation and reduction of antioxidant enzyme, attack biomolecules such as DNA and Phospholipids (Hinson et al., 2004).

 

In spite of phenomenal growth of modern medicine, there are no synthetic drugs available for the treatment of hepatic disorder.

However there are several herbs/herbal formulation claimed have possess beneficial activity in treating hepatic disorders. Natural products and their active principles as sources for new drug discovery and treatment of diseases have attracted of attention in recent years. Tylophora asthmatica a wild indigenous plant, belongs to the family Asclepidaceae and is commonly called as Indian ipecac. It is an endangered plant species, endemic to the state of Tamilnadu in India. The powdered leaves, stems and root contains 0.2-0.3% alkaloids, of these, tylophorine, tylophorinine are important alkaloids, various studies have confirmed the antiinflamatory stimulate of adrenal cortex (Udupa et al., 1991) anti asthmatic (Shivpuri et al., 1972) and the treatment of bronchitis, rheumatism and dermatitis (Nadkarri, 1976)

The aim of the present study was to evaluate the prevention role of

Tylophora asthamatica against acetaminophen induced lipid peroxidation  in hepatic damage.

 

MATERIAL AND METHODS:

Collection and processing of plant materials and preparation of the plant extract.

Collection and processing of plant material:

The fresh leaves of Tylophora asthmatica were collected during the month of February , 2008 in the Banks of Cauvery River, Tiruchirappalli, South India. It was botanically identified and authenticated. A voucher specimen (TAL-12) has been kept in our laboratory for future references. The leaves were shade dried, powdered, sieved through 410 mesh and stored in a tightly closed container for future use.

 

Preparation on of plant extract:

The powdered plant material (500) was extracted with petroleum ether (60-80oC) using soxhelt apparatus to remove lipids. It was filtered and the filtrate was discarded. The residue was extracted with methanol by soxhlet apparatus. The extract was completely dried in vauco, stored in refrigerator at 4oC and protected from sunlight until the time for extract administration. The yield of methanolic dried extract was 8.63% (w/w).

 

Animals used:

Adult healthy male wistar strain rats weighing 180 to 200g were procured from Fredrick Institute of Plant Protection and Toxicology, Padappai, Chennai, India. They were maintained under uniform laboratory conditions in standard steel cages and provided with food (Hindustan Lever Ltd., Bangalore) and water ad libitum. The animals were maintained under laboratory conditions for 2 weeks to acclimatize before performing the experiment.

 

Experimental protocol:

Paracetamol (Acetaminophen; Sigma Chemical Company, USA) was suspended in saline solution in boiling water and was administered intraperitoneally (i.p) after cooling (37oC) at a dose for 1g kg-1 body weight. This dose is known to cause liver damage in rats (Akintonwas et al., 1990). The experimental animals were divided into four groups, each group comprising six animals.

Group 1:         Control rats fed with standard diet.

Group 2:         Intoxication with 2ml of paracetamol suspension (1g kg-1 body weight, i.p.,)

Group 3:         Treatment with META extracts (200mg kg-1       body weight for 30days).

Group 4:         Co-treatment with META extracts (200mg kg-1 body weight  for 30 days; orally) prior to induction of liver damage with 2 ml of paracetamol suspension (1g kg-1 body weight, i.p.,).

 

Biochemical analysis:

The collected serum and liver tissue samples were used for the analysis of lipid peroxides [Ohkawa et al., 1979] ferritin [Drysdale and Munro, 1965], Iron [Ramsay, 1958], Total cholesterol [Parekh and Jung, 1970] and Phospholipids (Barttelle, 1959), total lipid (Folch et al.,1970).

 

Statistical analysis:

In the present investigation, the analysis tests whether changes among the groups differ significantly. Single way of analysis of variance was used to examine if significant differences existed in the different groups, all various are expressed as mean + SD for six rats in each group. The entire analyses were computed with the help of SPSS package (16 versions).

 

RESULT AND DISCUSSION:

Table (1) shows the level of lipid peroxides in both serum and liver tissue and level of iron and ferritin,total lipid, total cholesterol, phospholipid in the control and experimental groups of animals. Results indicate that the levels of lipid peroxides in both serum and liver and the levels of hepatic iron, were significantly increased (P<0.05) where as the levels of liver ferritin were significantly decreased (P<0.05) in the rats given with paracetamol (group II) compared to control rats, thereby showing the severity of oxidative stress induced as a result of the administration of the paracetamol, lipid peroxides are the prescriptive markers for free radical generation and development of oxidative stress (Sree priya et al., 1998).

 

The decreased level of lipid peroxide in group IV rats leads to the inference that META contracts the abnormal increase in lipid peroxides induced by acetamninophen, this might due to the antioxidative nature of the plant. The level of total lipid, total cholesterol in the liver significantly increase (P<0.05) in the group II rats except the level of phospholipids, which showed a significant decrease (P<0.05) compared to control rats. Rats treated with plant extract to the administration of overdose of paracetamol showed a restoration of the altered lipid levels induced by paracetamol toward near normal to thereby showing the modulating effect of the extract. Earlier studies have shown that excess level of the ROS formation by NAPQI leads to enhancement of lipid peroxidation and attack of biological molecules such as DNA and Phospholipids (Hinson et al., 2004).Thus Tylophora asthmatica to prevent lipid peroxidation  in acetaminophen induced hepatotoxicity.This needs further confirmation with the enzyme activities and histopathological studies.s

 

 

 

Table: 1 Level of serum lipid peroxides and Iron, ferritin, lipid peroxides in liver of control and experimental groups of rats

Parameters

Group I

Group II

Group III

Group IV

Serum lipid peroxide (n moles of MDA/mg protein)

1.29±0.02

2.41±0.02a*

1.31±0.03

1.84±0.04b*

Lipid peroxide n moles of MDA/mg protein

1.68±0.03

2.87±0.02 a*

1.71±0.03

2.07±0.02 b*

Iron (μg/g)

150.46±0.63

284.99±7.15 a*

152.13±0.33

164.24±0.37 b*

Ferritin (μg/g)

30.37±0.61

22.40±0.43 a*

31.01±0.54

28.94±0.57 b*

Total lipid (μg/g)

58.62±0.32

72.14±0.32 a*

58.91±1.26

60.02±0.81 b*

Total cholesterol (mg/g)

11.26±0.43

17.63±0.63 a*

10.97±0.67

12.07±0.61 b*

Phospholipid (mg/g)

22.34±0.15

18.84±0.13 a*

20.81±0.21

21.04±0.85 b*

Values are expressed mean ± SD for six animals. a As compound with group I. b As compared with group II. I each group. P<0.05.

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Received on 05.02.2011          Accepted on 02.03.2011        

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Asian J. Res. Pharm. Sci. 1(3): July-Sept. 2011; Page 71-73