Hepatoprotective Effect of Pergularia daemia on Cadmium Chloride Induced Toxicity in Rats

 

P. Vinoth Kumar1*, Ch. Sudheer Kumar2, V. Veera Thamarai Selvi1 and A. Amala Bricey3

1Department of Biotechnology, J.J college of Arts and Science, Pudukottai, India.

2School of Life Sciences, Department of Biotechnology, University College of Engineering, JNTU, Kakinada, Andhra Pradesh, India.

3Department of Biotechnology, Bharathidasan University, Trichy, India.

 

ABSTRACT:

The present study was undertaken to examine the inhibitory effect of the Pergularia daemia on cadmium chloride induced hepatoprotective activity in liver. In rats injected with cadmium chloride, the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), γ - glutamyl transferase (GGT) was significantly (p < 0.05) increased when compared to those values in control rats. The rats administrated with P.daemia extract and cadmium chloride showed a significantly (p < 0.05) decreased levels of serum AST, ALP, LDH and GGT. This study suggested that P.daemia may be implicated as a hepatoprotective agent for therapeutic purposes.

 

KEYWORDS: Cadmium chloride, Hepatoprotective activity, Pergularia daemia.

 

INTRODUCTION:

It is now well realized that environmental problems have increased exponentially in recent decades mainly because of rapid growth in human population and increased demand for several household materials. The toxic chemicals discharged into air, water and soil get into food chain from the environment. By entering into the biological system they disturb the biochemical processes leading to health abnormalities. Trace elements were known to have a variety of important biological functions and in many instances; they may have adverse effects on biological system1, 2, 3. Cadmium, a heavy metal well known to be highly toxic to both human and animals, is distributed widely in the environment due to its use in various industries. Some of the toxic effects of cadmium exposure are testicular atrophy, renal dysfunction, hepatic damage, hypertension, central nervous system injury and anemia4.

 

Pergularia daemia is distributed in the tropical and sub tropical area. In India it is very commonly found in hedges through cut most of cenfry to an altitude about 1000m in Himalayas and 900m in Southern India. Pergularia daemia known as “Veliparuthi” in Tamil, “Uttaravaruni” in Sanskrit. Phytochemically the plant has been investigated for cardenolides, alkaloid and saponins 5. The plant was found to contain various triterpenes and steroidal compounds 6. Dried leaf used as an emetic and bronchitis 7, antirheumatic 8, amenorrhea, dysmenorrheal 9, asthma 10. Other traditional uses of P. daemia include treating flatulence (gas), regulating body temperature and blood sugar, promoting digestion and improving mental processes.

 

 

Our study is mainly focused to explore the protective role of P. daemia extract on Cadmium chloride induced toxicity in rats by analyzing their biochemical alteration.

 

MATERIALS AND METHOD:

Experimental Animals:

Healthy male adult albino rats (Wistar strain) 6-7 weeks old, weighing 100±20 g was procured from ‘‘Sri Venkateswara Enterprises’’, Bangalore, India. They were housed in clean sterile polypropylene cages with proper aeration and lighting (12±1 hr day/ night rhythm) throughout the experimental period. During the course of the experiment, the temperature was maintained between 27°C. The animals were fed with commercially available pelleted rat feed (Gold-Mohur, M/S Hindustan Lever Ltd, Mumbai, India) during the acclimatization period and water ad libitum. The usage and handling of experimental rats followed the rules and regulation given by the Institutional Ethics Committee.

 

Plant material:

The plant sample of Pergularia daemia. Forssk were collected from Medicinal Garden of J.J College of Arts and Science, Pudukkottai district, Tamil Nadu, India. The collected plant sample were identified and confirmed with the help of Gamble flora of Tamilnadu. The plant sample was identified as Pergularia daemia. Forssk under the family of Asclepiadaceae. The large group of Asclepiadaceae consists of numerous members of medicinal plants. Among these Pergularia daemia. Forssk is a traditionally important medicinal plant.

 

Preparation of extracts:

The leaves of P. daemia were dried under shade, powdered with a mechanical grinder and passed through sieve no 40. The sieved powder was stored in airtight container and kept in room temperature until further study. The dried powdered material (500g) was extracted with 95% ethanol using soxhlet apparatus for about 48 hr. The aqueous extract was prepared by cold maceration (72 hr) and the solvents were removed from the extracts under reduced pressure using rotary vacuum evaporator.

 

Drugs and Chemicals:

Cadmium chloride was purchased from Sigma chemical Co. (St. Louis, MO, USA). The rest of the chemicals and biochemical were obtained from local firms (India) and were of analytical grade.

Treatment Schedule:

Male albino rats of Wistar strain selected for the study were acclimatized for 15 days. Then the rats were randomized into 4 groups, each consisting of six rats.

Group I: Normal rats (Fed with normal diet and water ad libitum).

Group II: Animals were orally treated with the leaf extract of P.daemia at a dose of 250 mg/kg body weight for a period of 15 days.

Group III:  Animals received subcutaneous administration of cadmium chloride at a dose of 1.25 mg/kg body weight throughout the experimental period for 15 days.

Group IV: Animals received subcutaneous administration of cadmium chloride (1.25mg/kg body weight) and orally treated with P.daemia leaf extract (250 mg/kg body weight) for a period of 15 days.

 

Estimations:

Blood was collected in unheparinised tubes and serum was separated for the estimation of marker enzymes such as AST, ALT, LDH and GGT. AST and ALT were estimated by the method of Reitmann and Frankel method 11, Lactate dehydrogenase and γ -glutamyl transferase by literature methods12, 13.

 

Statistical analysis:

All data were expressed as mean ± standard deviation of number of experiments. The Statistical significance was evaluated by one-way analysis of variance (ANOVA) using SPSS version 9.0 (SPSS, Cary, NC, USA) and Duncan’s multiple range test (DMRT) 14. A value of p < 0.05 was considered to indicate a significant difference between groups.

 

RESULTS:

The activities of liver AST, ALT, LDH and GGT specific markers in serum is shown in table-1. In cadmium chloride injected rats (Group-III), the activities of AST, ALT, LDH and GGT were significantly (p < 0.05) increased when compared to those values in control rats (Group-I). The rats administrated with cadmium chloride and P.daemia extract (Group-IV) showed a significantly (p < 0.05) decreased levels of serum AST, ALT, LDH and GGT. Administration of P.daemia extract significantly reversed the cadmium chloride induced changes in circulation towards near normal.

 

 

 

Table.1: Effects of P.daemia on the activities of hepatic serum markers in control and experimental rats

Sl. no

Groups

AST (IU/L)

ALT (IU/L)

LDH (IU/L)

GGT(IU/L)

1

Control

60.20±6.95

63.12±7.43

45.52±3.91

07.84±0.74

2

P.daemia extract

60.19±6.45

63.10±9.75

45.54±4.20

07.56±0.52

3

Cadmium chloride

120.13±11.46

145.30±11.50

102.16±06.90

12.02±01.94

4

Cadmium chloride + P.daemia extract

97.12±9.01

82.6±9.59

60.06±3.59

8.40±0.46

Values are mean± SD for six rats in each group.

 

 

DISCUSSION:

Cadmium is one of the most dangerous occupational and environmental toxins. It is found in drinking water, atmospheric air and even in food. Products of vegetable origin are the main carrier of cadmium compounds in food15. These characteristic features of Cadmium-induced liver toxicity were similar to those previously reported by other investigators16,17. Several mechanisms have been suggested for the induction of Cadmium-associated hepatotoxicity 16. The enzymes AST, ALT, LDH and GGT and are found in the liver and have been widely used for diagnostic purposes. AST is present in both the mitochondria and cytosol of liver cells, while (ALT) is found in the cytosol only. Liver cells damage releases these enzymes into the extracellular fluid and results in increased plasma levels of transaminase activity. Prolonged destruction of the hepatic cells results in more hepatic release to exacerbate hepatic dysfunction and causes an elevation in the serum levels of LDH which is in accordance with the results of our study18. Glutamyl transferase (GGT) activity is considered to be one of the best indicators of liver damage. Oral administration of cadmium chloride to rats resulted in a statistically highly significant increase in the levels of these enzymes: AST, ALT, LDH and GGT in the serum when compared with the control group received normal saline. On the other hand, Administration of P.daemia with the cadmium restored the levels of these enzymes in the serum of the rats as an indication of protective effect of P.daemia against liver damage induced by cadmium. An improvement of the parameters of the present investigation may be due to the decrease of cadmium accumulation in liver in the rats receiving P.daemia in addition to cadmium. Among the possible mechanisms, it might occur that P.daemia reduced the uptake of cadmium in liver by competition for a common transporter and demonstrates protective actions against the damages of hepatocytes function during cadmium intoxication in the rats.

 

CONCLUSION:

Our results indicate that P.daemia have a protective effect against cadmium induced toxicity in rats.

 

ACKNOWLEDGEMENT:

Authors are thankful to J.J College of arts and science for providing the facilities for me. I would thank to Miss. Jone Sylvia Dass, Miss. Jenifer, Miss. Jebi, for encouraging me to do my project. I would like to thank J. Arul Joshva, J. Crish i-bell for providing fund to me carrying out my project.

 

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Received on 16.11.2010

Accepted on 03.12.2010     

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Research J. Pharmacology and Pharmacodynamics. 3(1): Jan. –Feb. 2011, 19-21