Studying the Effect of the Ziziphora tenuior L. Plant on Some Biochemical Factors of Serum in Rats

The Ziziphora tenuior L. is an herbaceous, plant rich in essential oil. Ziziphora tenuior L. has antibacterial and disinfection effect and its vapour is used to treat respiratory tract problems and removing symptoms of the common cold. This plant is carminative, promotes expelling of phlegm and sputum and is a tonic for the stomach. To perform this study some chemical factors (triglyceride, cholesterol, HDL, LDL, ALT, AST and total protein) of the rat's serum were measured in the clinical laboratory. In this study, number of 24 white male wistar rats with the weight range of 215 ± 15 g and at 10 weeks of age were prepared from the Laboratory Animals Breeding Center of the University. Rats were divided into three classes of 8 and 200 mg/kg and 400 mg/kg of the Ziziphora tenuior L. plant essence was used in groups 1 and 2 respectively and no plant compound was used in the third group as the control group. The plant was prescribed for 21 days in groups. Amount of the cholesterol, ALT and triglyceride factors showed the significant difference compared to the control group.


Introduction
The Ziziphora tenuior L. (named as Kakoti in Persian) is a herbaceous, seasonal, annual, flocculent, slender and erect plant with a height of 5-15 cm (In vitro propagation of the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant activity).This plant has limited growth and is not able to resist dry condition of the summer.The flowering season of this plant is April-Jun (Verdian-Rivi, 2008;Al-Rawashdeh, 2011;Ghassemi et al., 2013).
There is abundant oil essence in the Ziziphora tenuior L. Researchers have concluded in studying compounds of the Ziziphora tenuior L. plant essence that three compounds of Pulegone (82.6%),Limonene (6.8%) and Cineol (1.9%) have had the highest percent and totally have constituted 91.36% of the essence (Al-Rawashdeh, 2011;Ghassemi et al., 2013;Karimi et al., 2013).Leaves and blooming branches of the plant (fresh or dried) are consumed as infused or sodden with water or tea (Amanlou et al., 2005).
Ziziphora tenuior L. is known as a medicinal herb, because its extract shows antifungal and antibacterial effects.Even a lot research studies have been performed on Ziziphora tenuiro, the research assay descripted in this manuscript is quite novel.Measuring the concentrations of chemical factors in serum is easy and relatively cheap.It might be helpful to find new usage of Ziziphora tenuior.Ziziphora tenuior L. has antibacterial and disinfection effect and its vapour is used to treat respiratory tract problems and removing symptoms of the common cold.Also, it is used as antispasmoic in gastrointestinal disorders.This plant is carminative, promotes expelling of phlegm and sputum and is a tonic for the stomach.Mixture of its used and powder is used in some areas in remedy of diarrhea.Its seed in used to treat fever in India.It is also useful to increase the sexual force (Naeini et al., 2005;Talebi et al., 2012;Ghassemi et al., 2013).
Infusion of the Ziziphora tenuior L. is useful in treatment of the rheumatoid arthritis and rickets.Also it relieves the chronic tonsillitis.This plant is a common antiepileptic agent and increases secretion of bile therefore, it is useful for liver.Infusing some grams of this plant in the hot water is useful to treat Stomachache, Bronchitis, Pertussis, bloodshed, etc. (Sharafzadeh & Alizadeh, 2012;Talebi et al., 2012;Ghassemi et al., 2013;Karimi et al., 2013;Tabatabaei et al., 2013).Habitats of this species have high potential to apiculture the due to the suitable nectar (Al-Rawashdeh, 2011;Darbandi et al., 2013;Ghassemi et al., 2013).To perform this study some chemical factors of the rat's serum were measured in the clinical laboratory.

Material and Method
This study is experimental and is conducted in 2014 in the Islamic Azad University of Shahrekord Branch.To perform this study some chemical factors (triglyceride, cholesterol, HDL, LDL, ALT, AST and total protein) of the rat's serum were measured in the Al-Mahdi clinical laboratory of Shahrekord.
Samples were collected from leaves of the Ziziphora tenuior L. plant and essence making was performed in the Islamic Azad University of Shahrekord Branch laboratory and they were accurately identified and confirmed by means of adapting the herbarium specimens.Then they were dried at 25-35 °C temperature for 3 hours.After drying they were crushed and extracted using BP (British Pharmacopoeia, 1988).It should be noted that 1 cc essence were obtained per 100 gr plant (British Pharmacopoeia, 1988).
In this study, number of 24 white male wistar rats with the weight range of 215 ± 15 gr and at 10 weeks of age were prepared from the Laboratory Animals Breeding Center of the Islamic Azad University of Shahrekord Branch and then were maintained in standard cages and ready access to water and food was provided for them.Rats were divided into three classes of 8 and 200 mg/kg and 400 mg/kg of the Ziziphora tenuior L. plant essence was used in groups 1 and 2 respectively and no plant compound was used in the third group as the control group.
The plant was prescribed for 21 days in groups.After 21 days animals were anesthetized using the chloroform in special containers provided for this purpose and blood sample was collected using the cardiac puncture technique (Barreto et al., 2008) and its serum was isolated using centrifuge apparatus at speed of 2000 cycle per minute and then AST, ALT, HDL, total protein, cholesterol and triglyceride factors were measured.Data were statistically analyzed using the SPSS software and significant levels (p < 0.05).

Results
Results were compared with each other using the Dunnet test.Amount of serum triglyceride was 45.3 ± 20.9 mg/dl in the group 1 that did not show significant difference compared to the control group (111.7 ± 21 mg/dl).Also amount of triglyceride in the group 2 (41.2 ± 18.2 mg/dl) was that did not show significant difference compared to the group control (111.7 ± 21 mg/dl) (Table 1).
Amount of total protein in the group 1 was (7.6-0.40 mg/dl) that has not significant difference compared to the control group (7.5 ± 1.7 mg/dl).Amount of total protein in the group 2 was (6.7 ± 1.3 mg/dl) that has not significant difference compared to the control group (7.5 ± 1.7 mg/dl) (Table 1).
Amount of the cholesterol in the group 1 was (65.4 ± 9.5 mg/dl) that showed the significant difference compared to the control group (142.2 ± 24.13 mg/dl).Also amount of the cholesterol in the group 2 was (60.6 ± 8.5 mg/dl) that showed the significant difference compared to the control group (142.2 ± 24.13 mg/dl) (Table 1).
Amount of the AST in the group 1 was (166.2±74.6 mg/dl) that did not show significant difference compared to the group control (173.7 ± 55.8 mg/dl).Amount of the AST in the group 2 was (124.2 ± 15 mg/dl) that did not show significant difference compared to the group control (173.7 ± 55.8 mg/dl) but in two groups show a relative decrease compared to the control group (Table 2).Amount of the LDL in the group 1 was (5.6 ± 3.4 mg/dl) that did not show significant difference compared to the group control (4.3 ± 1 mg/dl).Amount of the LDL in the group 2 was (3.3 ± 0.17 mg/dl) that did not show significant difference compared to the group control (4.3 ± 1 mg/dl) (Table 1).Amount of the HDL in the group 1 was (9.7 ± 2.01 mg/dl) that did not show significant difference compared to the group control (12 ± 1.4 mg/dl).Amount of the HDL in the group 2 was (10.2 ± 1.7 mg/dl) that did not show significant difference compared to the group control (12 ± 1.4 mg/dl) (Table 1).
Amount of the ALT in the group 1 was (36.3 ± 17.1 mg/dl) that showed the significant difference compared to the control group (80 ± 10.1 mg/dl).Also amount of the ALT in the group 2 was (33.6 ± 18.3 mg/dl) that showed the significant difference compared to the control group (80 ± 10.1 mg/dl) (Table 2).Amount of the AST in the group 1 was (166.3 ± 74.6 mg/dl) that did not show significant difference compared to the group control (173.7 ± 55.8 mg/dl).Amount of the AST in the group 2 was (124.2 ± 15 mg/dl) that did not show significant difference compared to the group control (173.7 ± 55.8 mg/dl) (Table 2).There is no significant difference in numbers with similar letters (p < 0.05).There is no significant difference in numbers with similar letters (p < 0.05).

Discussion
With regard to obtained results, the triglyceride and cholesterol levels in treatment groups showed significant decrease than the control group (p < 0.05) that this decrease was more significant in the group that received more amount of the essence.Therefore it can be concluded that the effect of the Ziziphora tenuior L. essence on level of the triglyceride and cholesterol factors depends on the dosage.The Ziziphora tenuior L. plant contains the essential oil that its ingredients are thymol and Carvacrol.Carvacrol decreases the plasma triglyceride concentration (Lee et al., 2003).Case et al. (1995) showed that Thymol and Carvacrol at doses of 0.15 (V/V) decreased the serum cholesterol in Leghorn chickens (Case et al., 1995).Elson and Qureshi (1995) showed that hypocholesterolemic effects of the Thymol and Carvacrol is related to the inhibition of the enzyme HMG_COA reductase (Elson & Qureshi, 1955).Tsherich (2000) reported that using the Carvacrol stimulates the lactobacillus reproduction and growth.Lactobacillus has the important role in improvement of blood factors and decreasing serum lipids (Tschirch, 2000).Also perkival reported that lactobacilli can metabolize and absorb the cholesterol in the Small intestine and decrease its absorption through the blood (Percival, 2001).Cerig (1999) also reported role of medicinal plants and effective oils in decreasing cholesterol and protecting against the cancer (Craig, 1999).Also the serum cholesterol concentration was shown by adding the effective oils in the livery of broilers (Case et al., 1995) that obtained results in conducted research are consistent with the present study.Youshika et al. (2000) also reported that plant essential oils can decrease the abdominal fat deposition.This decrease can be caused by reduction of serum lipids (Yoshioka et al., 2000).Phenolic compounds available in medicinal plants control activity of the (HMG_COA) enzyme and in result the cholesterol synthesis is controlled.This act increases LDL receptors in surface of liver cells and consequently the LDL catabolism is accelerated.Inhibitors of HMG_COA reductase decrease the LDL and the plasma triglyceride concentration to a lower concentration (Barreto et al., 2008).In the present study a relative decrease was observed in the LDL amount in the group receiving the 400 mg/kg doses of the Ziziphora tenuior L. essence.In this study enzymes serum ALT and AST were used to investigate the liver performance biochemistry.These enzymes naturally are existed in liver cells and these enzymes are released into the blood due to disorder in the plasma membrane or cells deposition when liver cells are damaged and so serum levels of these enzymes are increased.Therefore increased concentration of these two enzymes is considered as a suitable criterion for evaluating the level of the damage to liver cells (Chidambarama & Carani, 2010).In the conducted study the ALT level in both treatment groups showed the significant decrease than the control group that this decrease was more in the second group that had received 400 mg/kg of the plant essence.So it can be said that the plant essence depends on the dose in causing the decrease in ALT and has no toxic effects on liver cells in doses of 200 and 400 mg/kg but improve the liver activity.Also the AST level in both treatment groups showed the relative decrease than the control group.

Table 1 .
Studying effect of the Ziziphora tenuior L. plant on biochemical factors of the serum (LDL, HDL, total protein, cholesterol, total protein and triglyceride) in rat for 21 days

Table 2 .
Studying effect of the Ziziphora tenuior L. plant on liver enzymes (ALT, AST) in rat for 21 days