Effect of thyme (Thymus Vulgaris l.) oil on some biochemical parameters of diabetic female rats

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World Journal of Pharmaceutical Sciences ISSN (Print): 2321-3310; ISSN (Online): 2321-3086 Published by Atom and Cell Publishers All Rights Reserved Available online at: http://www.wjpsonline.

1 World Journal of Pharmaceutical Sciences ISSN (Print): ; ISSN (Online): Published by Atom and Cell Publishers All Rights Reserved Available online at: Original Article Effect of thyme (Thymus Vulgaris l.) oil on some biochemical parameters of diabetic female rats Rasha J. Tuama Department of Chemistry, College of Science, University of Thi Qar, Iraq ABSTRACT Received: / Revised: / Accepted: / Published: The present study aimed to evaluate the influence of thyme oil on some biochemical parameters of diabetic female rats. Twenty four female rats divided into four groups each group contain six animals as following: the first group I (control group) was treated with (0.4mL/kg) of normal saline for two weeks, the second group II (diabetic group) was treated with alloxan (125 mg/kg), the third group III (thyme oil) was treated with (0.4mL/kg) of thyme oil for two weeks and the fourth group IV (diabetic-thyme oil group) was treated with alloxan (125 mg/kg) and then treated with (0.4 ml/kg) of thyme oil for two weeks. The results indicated a significant increase in the level of serum glucose, total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), very low density lipoprotein (VLDL), creatinin and uric acid in group (II) comparison with group (I). While, the level of high density lipoprotein (HDL) decreased significantly in group (II) comparison with group (I). There was a significant decrease (P<0.05) in the level of serum glucose, TC, TG, LDL, VLDL, creatinin and uric acid in group (IV) comparison with group (II). Also, the level of HDL increased significantly in group (IV) comparison with group (II). Key words: Thyme oil, Diabetes, Alloxan, Rats. INTRODUCTION Diabetes mellitus is a non-communicable metabolic disorder characterized by hyperglycaemia due to overproduction and underutilization of glucose [1,2]. Diabetes tends to damage cell membranes which results in elevated production of reactive oxygen species (ROS). The generation of ROS appears to play a critical role in the pathogenesis of diabetes mellitus [3]. Hyperglycemias associated with diabetes also increase the production of ROS and affects antioxidant enzymes and reaction [4,5]. International Diabetes Federation (IDF) estimated that were 34.6 million people with diabetes mellitus in the Middle East and North Africa, a number that will almost double to 67.9 million by 2035 if concerted action is not taken to tackle the risk factors fuelling the epidemic of diabetes throughout the region [6]. Diabetes mellitus is known to impair many physiological functions. Some reports claim that plant oils can reduce these alteration caused by diabetes mellitus. Thyme (Thymus Vulgaris L.) is an important medicinal plant which belongs to the Lamiaceae family; it has been used for centuries as spice, home remedy, drug, perfume and insecticide. [7,8]. In medicine, it is used as antibacterial, anti-fungal, antioxidative, expectorant, antiseptic, antithelminthic and antitusive as reported by other authors [9]. Reports indicate that the volatile oil of thyme are among the main essential oils used in the food industry and in cosmetics as preservatives and antioxidants [10]. Thymus Vulgaris essential oil is a mixture of monoterpenes. The main compounds of this oil are the nature terpenoid thymol and its phenol isomer carvacrol, which have antioxidative, antimicrobial, antitussive, expectorant, antispasmodic, and antibacterial effects [11]. Terpenoids, flavonoid glcones, flavonoids glycosides and phenolic acid were also found in Thymus Vulgaris L [12]. The present study aimed to investigate the effect of thyme oil on some biochemical indices of diabetic female rats. MATERIALS AND METHODS Thyme oil: It was obtained from the local markets in Al-Nassiriyah city, Thi-Qar province, Iraq. Oil has been keeping in a dark containers isolated from *Corresponding Author Address: Rasha J. Tuama, Department of Chemistry, College of Science, University of Thi Qar, Iraq; ; rashajasim17@yahoo.com

2 the air to protect it from moisture, heat and polymerization. Uric acid and Creatinine were measured according to [19, 20]. Induction of diabetes: After fasting of 18 hours, the rats were intraperitoneally injected with alloxan (BDH, England) at a single dose of 125 (mg/kg) of body weight dissolved in 1mL of normal saline [13]. After injection, the rats had free access to food and water. Diabetes was allowed to develop and stabilize in these alloxan-treated rats over a period of seven days. Diabetes mellitus was defined in these rats using determination of fasting blood glucose levels. The rats showed the fasting blood glucose more than 200 mg/dl were considered diabetic and selected for the experimentation. Experimental design: Twenty four mature female albino rats (Rattus norvegicus), weighting ( ) gm were used in the present study. The experimental animals were obtained from Department of Biology, Collage of Science, Thi- Qar University, Iraq. The animals were housed in standard plastic cages and maintained under standard laboratory conditions (12h light/ 12 h dark cycles, 22±2 C ) were fed on standard pellet and tap water ad libitum. The animals divided into four equal groups, each group into consist of (6) rats: 1. The first group (control group) treated orally with (0.4mL/kg) of normal saline for two weeks. 2. The second group (diabetic group) treated with a single dose (125 mg/kg) of alloxan. 3. The third group (thyme group) treated orally with (0.4mL/kg) of thyme oil for two weeks. 4. The fourth group (diabetic-thyme oil group) diabetic female rats treated with (0.4mL/kg) of thyme oil for two weeks. Blood samples collection: At the end of two weeks, the rats were fasted for 12 hours, euthanize under diethyl ether and sacrificed. Whole blood was collected by cardiac puncture with sterile needles, placed into sterile tubes without EDTA, clotted blood was centrifuged at 3000 rpm for 15 min, and then serum was separated and stored in freezer until the time of essay. Measurement of Biochemical Parameters: The used reagents were supplied by Biolabo (France). Serum glucose was measured according to [14], Serum total cholesterol (TC) was measured according to [15], Serum triglyceride (TG) was measured according to [16], serum high density lipoprotein (HDL) was measured according to [17] and measurement of low density lipoprotein (LDL) and very low density lipoprotein (VLDL) according to [18]. LDL and VLDL were measured as follows: [LDL = total cholesterol (HDL+VLDL)] and [VLDL = TG/5]. Statistical analysis: Statistical analysis was done using the software SPSS version 15, the result were expressed as mean ± standard deviation (means ± SD). One way ANOVA test was used to compare parameters in different studied groups. P-Values (P<0.05) were considered statistically significant. RESULTS AND DISCUSSION Effect of thyme oil on blood glucose of diabetic rats: Table (1) explained the effect of thyme oil on some biochemical parameters of diabetic female rats. The results indicated a significant increase (P<0.05) in the level of serum glucose of group (II) compared with group (I). While, there was nonsignificant differences (P<0.05) in the level of serum glucose of (III and IV) groups compared with group (I). On the other hand, there was a significant reduction (P<0.05) in the level of serum glucose of (III and IV) groups compared with group (II), figure (1). The results from the present study showed that injection of alloxan leads to significant increase of glucose level in rats. This increase due to destruction of β cells of pancreas by alloxan which lead to stop production of insulin [21]. Also causes diminishes of inters of glucose to cells leads to increase its level in blood [22]. Alloxan reacts with two SH-groups in the sugar binding side of glucokinase resulting in the formation of the disulfide bond and inactivation of the enzyme [23]. Thyme oil is rich in flavonoids and phenolic compounds, especially thymol and carvacrole [24]. This may be due the reduction in blood glucose by thyme oil in groups (III and IV) compared with group (II) to the action of carvacrole or thymol which is an insulin mimetic and possibly the ability of the plants oil to alter the inhibitory activity of alloxan on glucokinase which is the glucose sensor of the β cells [25,26]. Effect of thyme oil on lipids profile of diabetic rats: The results indicated a significant increase (P<0.05) in TC, TG, LDL and VLDL levels of group (II) compared with group (I). While, there was a significant decrease (P<0.05) in HDL level of group (II) compared with group (I). The results showed a significant decrease (P<0.05) in TC, TG, and VLDL levels of group (III) compared group (I). Also, there was non-significant differences (P<0.05) in TC level of group (IV) compared with group (I) and there was a significant increase (P<0.05) in TG and VLDL levels of group (IV) compared with group (I). While, there was nonsignificant differences (P<0.05) in HDL and LDL levels of groups (III and IV) compared with group (I). On the other hand, a significant reduced 321

3 (P<0.05) in TC, TG, LDL and VLDL levels was observed in groups (III and IV) compared with group (II). While, the HDL level increased significantly (P<0.05) in groups (III and IV) compared with group (II), table (1), figures (2, 3, 4, 5 and 6). Thyme oil was associated with lower serum levels of TC, TG, LDL and VLDL but higher serum levels of HDL [27]. The decline in the level of cholesterol perhaps due to the thyme oil contain flavone and characterized by possessing antioxidant qualities and depressor of blood lipids [26]. Many studies investigated that TG accumulation in tissues causes insulin resistance and induces impairment of pancreatic beta-cell function. A reduction in TG led to reduction in body weight and intrabdominal fat weight, decreases in plasma TG, insulin and glucose levels and decrease in the TG secretion, decreases in the TG content in the liver, pancreas and muscles, improvement of the glucose infusion rate and improvement of pancreatic beta-cell function [28]. The decline in the level TG has been attributed to the role of thyme oil antioxidant where diabetes is one of the factors that cause oxidative stress events [29]. The increase in the level of VLDL may be due to the low level of insulin as it reduced enzyme activity to lipoprotein lipase responsible for removing triglycerides [30], and as a result of the high level of triglycerides in the blood increases the level of VLDL for being contain a large proportion of triglycerides. As well as the diabetes that leads to a shortage in receptor VLDL in the cells which entry in to the cell and thus higher concentration in the blood [31]. The decline in the level of VLDL this fact reduces the level triglyceide in the blood level drops so for being VLDL contain a low percentage of triglycerides [32]. The rise in the level of LDL may be due to change in the level of apolipoprotien which plays an important role in maintaining concentrations of natural LDL and HDL in the blood, and could be due to the low level in insulin [33]. The decline in the level LDL has been attributed to the thyme oil prevents process lipid peroxidation through possession of the effectiveness of antioxidants and thus prevent the decomposition of fat and decrease in the LDL level [34]. Effect of thyme oil on kidney functions of diabetic rats: The results showed a significant increase (P<0.05) in the levels of creatinine and uric acid in group (II, III and IV) compared with group (I). While, there was a signification decrease (P<0.05) in the levels of creatinine and uric acid in groups (III and IV) compared with group II. On the other hand, there was non-signification differences (P<0.05) in creatinin between group III and IV, figures (7 and 8). The diabetic hyperglycemia induces the elevation of plasma creatinine and uric acid in diabetic rats, which are considered a significant marker of renal dysfunction [35]. On the other hand, treatment of the diabetic rats with thyme oil caused reduction in the serum creatinine and uric acid levels in group (IV) compared to the mean values of diabetic group. This may be due to the presence of polyphenols and flavonoids in thyme might be responsible for the antioxidant activities and the reduction of serum creatinine and uric acid levels [36]. Conclusion Thyme (Thymus Vulgaris L.) has antihyperglycemic and antilipidemic effect, and kidney functions improvement, which may be help in treatment of diabetes and diabetes complications. Table 1: Effect of thyme oil on some biochemical parameters in diabetic rats Animal groups I II III IV Glucose ± 8.44 b ± a ± 4.51 b ± 9.90 b TC 94.34± 1.64 b ± 3.86 a 87.65± 4.17 c 97.44± 3.23 b TG ± c ± 9.56 a ± 4.49 d ± 3.70 b HDL 41.93± 2.70 a 36.47± 3.41 b 44.34± 3.16 a 44.85± 2.82 a LDL 17.57± 5.14 b 31.64± 4.40 a 13.52± 6.35 b 14.68± 4.15 b VLDL 34.83± 2.89 c 40.14± 1.91 a 29.75± 0.89 d 37.90± 0.74 b Creatinin (μmol/dl) 85.33± 8.84 c ± 2.59 a 91.66± 4.19 b 94.78± 3.19 b Uric acid (μmol/dl) ± 19.1 d ± 6.85 a 165.6± c ± b * Each value represents (mean ± S.D) values with non identical superscript (a, b or c... etc.) were considered significantly different (P < 0.05). 322

4 Glucose TC Figure 1 Changes in the concentration of serum Glucose of animal groups Figure 2 Changes in the concentration of serum TC of animal groups TG HDL Figure 3 Changes in the concentration of serum TG of animal groups Figure 4 Changes in the concentration of serum HDL of animal groups 323

Uric acid (μmol/dl) Figure 7 Changes in the concentration of serum Creatinin of

5 LDL VLDL Figure 5 Changes in the concentration of serum LDL of animal groups Figure 6 Changes in the concentration of serum VLDL of animal groups Creatinin (μmol/dl) Uric acid (μmol/dl) Figure 7 Changes in the concentration of serum Creatinin of animal groups Figure 8 Changes in the concentration of serum Uric acid of animal groups 324

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