Hepatoprotective Effect of Opuntia Ficus-Indica Aqueous Extract against Carbon Tetrachloride-Induced Toxicity in Rats

Corresponding Author: Zouhir Djerrou Department of Sciences of Nature and Life, Faculty of Sciences, University of August 2


Introduction
Liver is the vital organ of metabolism and excretion. About 20000 deaths found every year were due to liver disorders. In addition, hepatocellular carcinoma is one of the ten most common tumors in the world with over 250000 new cases each year (Gupta and Misra, 2006). In spite of tremendous strides in modern medicine, there are hardly any drugs that stimulate liver function, offering protection to the liver from damage and helping regeneration of hepatic cells (Goyal and Sharma, 2012). Consecutively, several scientific studies have focused medicinal plants used in the different traditional systems, in the hope to find more potent drugs with minimum adverse effects.
Opuntia ficus-indica L. is a tropical or subtropical plant belonging to the Cactaceae family and is mainly used for fruit production (De Cortázar and Nobel, 1992).
It can be used also as a vegetable forage resource for livestock feed in arid and semiarid lands during periods of drought and shortage of herbaceous plants (Felker et al., 2006). In the majority of scientific medicinal researches involves the leaves (cladodes) rather than the fruit (El-Kossori et al., 1998). The cladodes are utilized, in traditional medicine, for the management of ulcers, wounds, rheumatic pain and fatigue. Experimental studies showed that cactus pear could reduce glucose and cholesterol levels in human blood (Frati et al., 1990;Stintzing et al., 2001). Chemopreventive effect on oxidative stress and genotoxicity was also recently investigated (Brahmi et al., 2011a). The chemical composition of cactus cladodes has been investigated in various studies. It is characterized by a high value of water (85-92%), 4-6% of carbohydrates, 1% proteins, 0.2% fats, minerals (1%), vitamin C (12.7 mg/100 g) and ß-carotene (12.9 µg/100 g) (El-Kossori et al., 1998;Rodriguez-Felix and Cantwell, 1988).
The aim of the present study is to evaluate the possible hepatoprotective effect of aqueous extract from cactus cladodes against carbon tetrachloride.

Animals
The study was conducted on 24 male Wistar rats (180-230 g weight). The animals were randomly divided on 4 groups of 6 each and were kept in standard cages for an acclimation period of 15 days with laboratory conditions. Food and water were provided adlibitum and a temperature of 22±2°C with a 12/12 h light/dark cycle were maintained.

Tested Drugs
Cladodes of Opuntia ficus-indica were collected in the region of Tamalous (East of Algeria) and the aqueous extract was prepared each 4 days.
CCl4 and ether were obtained from Faculty of Natural Sciences and Life of Constantine University.

Experimental Protocol
Four groups were formed, one control and 3 experimental lots, as follows: • Group I: Normal control (CRL), received 2 mL of distilled water orally for 15 consecutive days • Group II or CCl4: These rats were gaved every 3 days by CCl4 at a dose of 1.5 mL/kg for 15 days • Group III or OFI1: These rats received daily 2 mL/kg of Opuntia ficus-indica aqueous extract and carbon tetrachloride (CCl4) every 3 days at a dose of 1.5 mL/kg for 15 days via oral route • Group IV or OFI2: These animals received daily oral dose of 5 mL/kg of Opuntia ficus-indica aqueous extract and CCl4 every 3 days at a dose of 1.5 mL/kg for 15 days The administration of CCl4 was performed 60 min after oral gavage of rats with the aqueous extract of Opuntia ficus-indica. All animals were given using a stomach tube. The experimental procedures involving the handling and treatment of animals were approved by the ethical committee of the institute of veterinary sciences, University of Constantine 1, Algeria.

Biochemical Analysis
All survived rats until 15th day were anesthetized on the following day with ether and blood samples were collected by cardiac puncture into heparinized tubes and centrifuged at 3000 rpm for 10 min to collect sera. The biochemical analysis was performed in Biochemistry Laboratory (Polyclinic of Ain Smara, Constantine) including: ALT, AST, creatinine, urea and uric acid.

Statistical Analysis
All results were expressed as mean with variance. Data were analyzed using one-way ANOVA. The differences between groups were considered significant at p<0.05.

Results
The results recorded in Table 1 and Figs. 1 and 2 showed a significant elevation of ALT (p<0.05) and AST (p<0.001) enzymes in Group II due to CCl4 application compared with normal animals with no alliteration of other measured parameters. The administration of Opuntia ficus-indica at a dose of 2 mL/kg has resulted in a significant reduction of AST (p<0.001) in OFI1 group when compared to CCl4 group with no significant impact on the other tested parameters Figs. 3-5. However, the application of the aqueous extract at a dose of 5 mL/kg has accentuated the elevation of AST and ALT but nonsignificantly in the OFI2 group compared to CCl4 group.

Discussion
Carbon tetrachloride is a well documented hepatotoxic drug used to induce hepatic cirrhosis model for scientific research (Gutiérrez and Solís, 2009;Karandikar et al., 1963;Recknagel, 1983). In the present study, we have applied this toxic product via oral route each 3 days for a period of 15 days to ovoid a high rate of mortality. A study of Jang et al. (2008) has compared some experimental protocols to induce liver cirrhosis in mice and rats via intra peritoneal and oral routes using 3 hepatotoxic products. They have shown that livers from animals administered CCl4 orally twice a week for 10 weeks was the most effective to achieve sufficient fibrosis and greatest reproducibility with acceptable animal survival.
In the present study, the dose of 2 mL/kg of cactus aqueous extract has shown a hepatoprotective effect in term of AST decrease. However, the dose of 5 mL/kg has accentuated the hepatic lesions which suggest that this dose is very high and may be toxic for tested animals. In a review of Feugang et al. (2006), were investigated the phytochemical composition and the medicinal uses of cactus. This vegetable has shown anti cancer, anti viral, anti diabetic (type 2 diabetes), anti hyperlipidemic and anti hypercholesterolemic effects, anti oxidant properties, anti-inflammatory activity and hepatoproptective effect against CCl4-induced hepato-toxicity in rats. In a study of Brahmi et al. (2011b) conducted on mice, cactus cladode extract has proven effective activity in the protection against benzo (α) pyrene which is a widespread environmental genotoxin classified as probably carcinogenic in human inducing liver injury. A double-lind, placebo controlled, crossover trial of Wiese et al. (2004), conducted on 64 healthy volunteers, has concluded that Opuntia ficus-indica extract has a moderate effect reducing hangover symptoms, apparently by inhibiting the production of inflammatory mediators. Finally, a study of Ncibi et al. (2008) has shown that cactus stem extract protects the liver and decrease the toxicity induced by chlorpyrifos (organophosphorous pesticide).

Conclusion
In view of the present results, it may be concluded that Opuntia ficus-indica aqueous extract at a dose of 2 mL/kg exerted a hepatoprotective effect against carbon tetrachloride-induced toxicity in rats at least by decreasing AST activity.

Funding Information
This study was supported by a CNEPRU research project (I00920130067).

Author's Contributions
Zouhir Djerrou: Designed the research plan and organized the study, participated in all experiments, coordinated the data-analysis and has drafted the article.
Zineb Maameri: Participated in all experiments and data analysis.
Sihem Halmi: Participated in all experiments and acquisition of data.
Loubna Benmaiza: Participated in data analysis. Youcef Hamdipacha: Participated in analysis and interpretation of data.