Influence of Pistacia Lentiscus Oil on Serum Biochemical

European Journal of Scientific Research ISSN 1450-216X Vol.24 No.4 (2008), pp.591-600 © EuroJournals Publishing, Inc. 2008 http://www.eurojournals.com/ejsr.htm Influence of Pistacia Lentiscus Oil on Serum Biochemical Parameters of Domestic Rabbit Oryctolagus Cuniculus in Mercury Induced Toxicity Maarouf Tounes Laboratory of Animal Ecophysiology, Department of Biology, Faculty of Sciences University of Annaba, Annaba 23000, Algeria E-mail: albeled63@yahoo.fr Cherif Abdennour Laboratory of Animal Ecophysiology, Department of Biology, Faculty of Sciences University of Annaba, Annaba 23000, Algeria E-mail: cherifabdennour@yahoo.fr Nadjet Houaine Laboratory of the Analyses, Health Center, Sidi Ammar, Annaba 23000, Algeria Abstract The impact of Pistacia lentiscus oil, on mercury induced toxicity in domestic rabbit Oryctolagus cuniculus was investigated. Twenty four males were divided into three groups. The control was fed a basic diet, whereas the other two groups were treated either by Hg alone (1g HgCl2/Kg food) or Hg-oil (1g HgCl2/Kg food + 5% Pistacia oil), respectively, for 37 consecutive days. The following serum parameters were estimated; alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, uric acid, triglicerides, total cholesterol, HDL-cholesterol and LDLcholesterol. Compared to the control, results have revealed a significant decrease in serum ALP activity in mercury treated group, accompanied by a significant increase in serum AST. Furthermore, serum urea concentration was significantly higher in the mercury group compared to the control. However, the concentration of urea and the activity of ALP and AST of the Hg+oil group were not statistically different from the control. Moreover, no significant variations were recorded concerning AST, creatinine and uric acid. The lipid profile; triglycerides, total cholesterol, HDL-cholesterol and LDL-cholesterol were not significantly varied between the three groups, despite the observed elevated concentration of triglycerides in Hg+oil group. In conclusion, P. lentiscus oil may partially help in the protection against mercury intoxication as in the case of ALP, AST and urea, and it could also be considered a safe nutritional source, at least by maintaining total cholesterol and LDL-cholesterol in their normal ranges. Keywords: Hg, lipid profile, oil, Pistacia lentiscus, rabbit, serum biochemical parameters Influence of Pistacia Lentiscus Oil on Serum Biochemical Parameters of Domestic Rabbit Oryctolagus Cuniculus in Mercury Induced Toxicity 592 Introduction Food contamination by toxic trace elements is nowadays an evident health problem worldwide for general population (Endo et al., 2005). Mercury is the most dangerous trace element, which is originated from different anthropogenic sources. Due to its physico-chemical properties, it is found in the air, water, soil and food, polluting, however, the different ecosystems. Mercury is found to alter the physiological and the biochemical functions of living organisms (Tsalev, 1985), and cause a wide range of clinical symptoms in occupationally exposed workers (Abdennour et al., (2002). The toxic effects of Hg on human and animal health have been reported extensively (WHO, 1989). When binding to cell components, mercury provokes the oxidative stress, leading to the formation of a number of toxic substances. Thus, lipids are amongst the target molecules to be oxidised by mercury (Mahboob et al. 2001; Hussain et al., 1999). Consequently, mercury can alter membrane lipid structure and functions (Ganser & Kirschner, 1985) and even inhibits lipid synthesis in the nervous system (Cloez et al., 1987). It appears that the impact of some plant product used in the traditional medicine are needed to be studied, in order to know the physiopathologic states of human exposed to mercury. Such product is represented by the oil of Pistacia lentiscus, which is one of the commonly used tools in folk medicine in Algeria to cure varieties of diseases, especially some respiratory, digestive and skin disorders. However, this plant is widely distributed in north Algeria, and its oil is obtained and stocked in the majority of families. Oils are generally known to be one of the powerful antioxidants. The protective effect of plant and fish oil supplementation against oxidative stress in rat has been reported (Ando et al., 2000; Kikugawa et al., 2003). Furthermore, dietary oils rich on polyunsaturated fatty acids are found to be important factors in the risk assessment of mercury exposure (Jin et al., 2007; Jin et al., 2008; McVey et al., 2008). In the Mediterranean region, many studies on the chemical composition of P. lentiscus have been carried out so far (Wagne, 1999; Duru et al., 2003; Zrira et al., 2003; Vidrich et al., 2004; Benyoussef et al., 2005), but reports on its effects on experimental animals and human biochemical parameters are few in the literature (Al-Said et al., 1986; Janakat and AL-Merie, 2002; Triantafyllou, et al., 2007). The objective of the present work is therefore, to evaluate the beneficial roles of P. lentiscus oil, traditionally extracted, on serum biochemical parameters in mercury contaminated diet of domestic Oryctolagus cuniculus rabbit. Such parameters are generally representing the health status of the individuals. Materials and Methods 24 male rabbits Oryctolagus cuniculus aged 7 months old, with a mean weigh of ……….g were divided into 3 groups. The ration was composed of fresh forage and hay (50%), wheat bran (18%), corn bran (16%), salts and vitamins (AOAC, 1980). Animals were put in the animal house (Department of Biology, university of Annaba) under standard conditions of temperature, light and humidity and food was given ad libitum. The control was fed a basic diet, while the other two groups were treated either by Hg (1g HgCl2/Kg food) or Hg-oil (1g HgCl2/Kg food + 50g/Kg food of Pistacia oil). However, Pistacia lentiscus oil was collected from Taref region (north-east Algeria) and prepared by traditional method, where seeds were homogenised, mixed with warm water, filtered, and then centrifuged. After five weeks continuous treatments, animals were decapitated and blood was received in dry test tubes, and then centrifuged at 5000 rpm/15 minutes. The automate apparatus METROLAB 2300 (Random Access Clinical Analyzer) was used to measure serum alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, uric acid, triglycerides, total cholesterol, HDL-cholesterol and LDL-cholesterol. 593 Maarouf Tounes, Cherif Abdennour and Nadjet Houaine Statistical analysis has been carried out by student t-test to compare between paired groups, whereas the one way analysis of variance (ANOVA) was used to compare between groups. Results are expressed as mean ±SD and the statistical test was considered significant at p<0.05 level. Results Results of the serum enzymes ALP, AST, ALT, urea, creatinine and uric acid representing liver and kidney functions are shown in figures 1-6. The activity of ALP was significantly reduced in rabbits exposed to mercury alone compared to the control. Contrary, the serum AST activity was increased significantly in the mercury treated group compared to the control. Accordingly, serum urea concentration was also significantly increased in mercury group. When comparing between the three groups, ANOVA has revealed a significant variations in ALP, AST and urea. However, the AST, creatinine and uric acid were not significantly changed in all cases. Meanwhile, in the Hg-oil group, the ALP, ALT, AST, urea, creatinine and uric acid have not been varied significantly compared to the other two groups. Figure 1: Comparison of ALP activity (X±SD) in the serum of rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. Superscript letters differ significantly at p<0.05. a: G1 vs G2; d: G1 vs G2 vs G3 ALP 80 70 ACTIVITY (IU/L 60 50 40 30 20 10 0 G1 GROUPS G2 G3 a d Figure 2: Comparison of AST activity (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. Superscript letters differ significantly at p<0.05. a: G1 vs G2c: G2 vs G3; d: G1 vs G2 vs G3. AST 50 45 40 35 30 25 20 15 10 5 0 c d a ACTIVITY (IU/L) G1 GROUPS G2 G3 Influence of Pistacia Lentiscus Oil on Serum Biochemical Parameters of Domestic Rabbit Oryctolagus Cuniculus in Mercury Induced Toxicity 594 Figure 3: Comparison of ALT activity (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. a: G1 vs G2; b: G1 vs G3; d: G1 vs G2 vs G3; p<0.05. ALT NS 45 40 35 30 25 20 15 10 5 0 G1 GROUPS G2 G3 Figure 4: Comparison of urea concentration (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. Superscript letters differ significantly at p<0.05. a: G1 vs G2; d: G1 vs G2 vs G3. UREA 0,6 CONCENTRATION (g/L) 0,5 0,4 0,3 0,2 0,1 0 G1 GROUPS G2 G3 a d ACTIVITY (IU/L) 595 Maarouf Tounes, Cherif Abdennour and Nadjet Houaine Figure 5: Comparison of creatinine concentration (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. CREATININE NS CONCENTRATION (mg/L) 14 12 10 8 6 4 2 0 G1 GROUPS G2 G3 Figure 6: Comparison of uric acid concentration (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. URIC ACID NS 16 14 12 10 8 6 4 2 0 G1 GROUPS G2 G3 CONCENTRATION (mg/L) The results of the lipid profile; triglycerides, total cholesterol, HDL-cholesterol and LDLcholesterol are presented in figure 7-10. The most noticeable result was the triglyceride concentrations, which have been increased in the Hg-oil group, but it was not statistically significant compared either to the control, or to the mercury group. Figure 7: Comparison of triglycerides concentration (X±SD) in the serum of male rabbit in the control (G1), in Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. TRIGLYCERIDES NS CONCENTRATION (g/L) 2,5 2 1,5 1 0,5 0 G1 GROUPS G2 G3 Influence of Pistacia Lentiscus Oil on Serum Biochemical Parameters of Domestic Rabbit Oryctolagus Cuniculus in Mercury Induced Toxicity Figure 8: Comparison of total cholesterol concentration (X±SD) in the serum of male rabbit in the control (G1), Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. TOTAL CHOLESTEROL NS 0,8 0,7 0,6 0,5 0,4 0,3 0,2 0,1 0 G1 GROUPS G2 G3 CONCENTRATION (g/L) 596 Figure 9: Comparison of total cholesterol concentration (X±SD) in the serum of male rabbit in the control (G1), Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. HDL-CHOLESTEROL NS 0,4 0,35 0,3 0,25 0,2 0,15 0,1 0,05 0 G1 GROUPS G2 G3 CONCENTRATION (g/L) Figure 10: Comparison of total cholesterol concentration (X±SD) in the serum of male rabbit in the control (G1), Hg (G2) and in Hg+Pistacia lentiscus oil (G3) after 5 consecutive weeks. LDL-CHOLESTEROL CONCENTRATION (g/L) 0,3 0,25 0,2 0,15 0,1 0,05 0 G1 GROUPS G2 G3 NS 597 Maarouf Tounes, Cherif Abdennour and Nadjet Houaine Discussion Mercury contamination generates a prooxidative environment, leading to oxidative damage, which in turn could disturb some cell functions. However, in the mercury exposed rabbits, the activity of alkaline phosphatase was decreased, whereas that of AST was increased, with no change in ALT level. In this case, mercury may affect intestinal functions by inhibiting the enzyme activity during absorption processes; especially serum alkaline phosphatase is mainly originated from intestines, liver and bones. Accordingly, Bapu et al., (2003) have observed a decrease in the alkaline phosphatase in brain and spinal chord of mice exposed to organic mercury. On the other hand, the liver cell lyses, is probably responsible on the observed increase in AST activity, since liver is among the principal target organ for mercury intoxication. Mercury is one of the agents which disturb cell lipid membranes (Ganser & Kirschner, 1985), leading to the release of hydrolases (Bapu et al., 2003). It seems that P. lentiscus oil has a bouncing effect on alkaline phosphatase activity. Accordingly, boiled aqueous extract of P. lentiscus showed marked antihepatotoxic activity against carbon tetrachloride intoxicated rats by returning both serum AST and ALT to normal levels (Janakat and AL-Merie, 2002). A contradictory result was reported by Endo et al., (2005), where the activity of serum ALT of rat was increased, accompanied with no change in both AST and alkaline phosphatase in mercury contaminate whale red meat. In this study, the exposure to mercury has increased serum urea concentration after 37 days, confirming, however, the presence of renal functional disturbances, as a result of mercury accumulation, particularly mercury can cause anuria (WHO, 1991). It appears that the period of exposure to mercury could affect kidney functions differently. Thus after a short period of seven days, serum urea was reduced in rat given mercury contaminated whale red meat (Endo et al., 2005), while it increased in mice received a single dose of mercury during five consecutive days (Brandao et al., 2006). Urea augmentation in this study could also come from protein catabolism acceleration because of oxidative stress provoked by mercury. However, the supplementation of oil to rabbit diet, in this study, has kept urea level within its normal range. Accordingly, some P. lentiscus oil components are reported to be effective in protecting against chemically induced liver injury in laboratory animals (Liu, 1995; Rios et al., 2000). Serum triglycerides in the actual study have not been changed significantly either in the presence of mercury alone, or combined with of P. lentiscus oil. It is known that triglycerides are essential source of energy for cellular functions. Importantly, total cholesterol and LDL-cholesterol levels have not increased after P. lentiscus oil supplementation. To know that blood cholesterol and LDL levels are well-established risk factors for cardiovascular disease, while HDL is involved in reverse cholesterol transport, which reduces tissue cholesterol levels and may provide a protective effect (Gross, 2008). Accordingly, P. lentiscus oil was found to have antihyperlipidemic properties (Liu, 1995; Rios et al., 2000). Besides, P. lentiscus oil is characterised by its good nutritive quality where it contains 73% (Yousfi et al., 2002) and 69 % (Wagne, 1999) of essential polyunsaturated fatty acids (oleic + linoleic). Thus, linoleic acid, the most common form of polyunsaturated fatty acid is effective in reducing blood cholesterol concentrations, HDL-cholesterol and VLDL-cholesterol and lesser reduction of LDL-cholesterol (Gross, 2008). Generally, monounsaturated fats as olive oil have a neutral effect on cholesterol, whereas polyunsaturated fats as P. lentiscus decrease cholesterol levels (Katan et al., 1994). Moreover, triglyceride levels were slightly lower in the serum of rat fed for one month on a diet containing 10% P. lentiscus oil, collected from the same region (north-east Algeria), compared to 10% olive oil, but the concentrations of serum cholesterol were almost identical (Mezghache, 2002). Contrary, triglycerides, HDL, and total cholesterol serum levels increased with diet containing coconut oil, known by its high proportion of saturated fatty acids (Feoli et al., 2003). The brain is particularly sensitive to oxidative attack because of its high level of unsaturated lipids and high rate of oxidative metabolism (Goering et al., 2002), and mercury is one of the agent which inhibits lipid synthesis in the nervous system (Cloez et al., 1987), causing a reduction in tissue cholesterol levels (Sood et al. 1997). However, P. lentiscus oil could be used as a reserve to replace the damaged unsaturated fatty acids in cell membranes. Moreover, Jin et al., (2007) have confirmed the Influence of Pistacia Lentiscus Oil on Serum Biochemical Parameters of Domestic Rabbit Oryctolagus Cuniculus in Mercury Induced Toxicity 598 key role played by dietary fats when assessing organic mercury exposure. Nevertheless, strains and nutrition are one of the main factors which influence mercury toxicity (WHO, 1991). Besides, the significance of diet in mercury detoxification has been mentioned (Abdennour et al., (2002). 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