Hepatoprotective and Hypolipidemic Effects of Satureja Khuzestanica Essential Oil in Alloxan-induced Type 1 Diabetic Rats

Document Type: Research article

Authors

1 Department of Biochemistry, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran. Razi Herbal Researches Center, Lorestan University of Medical Sciences, Khorramabad, Iran.

2 Department of Anatomy, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.

3 Department of Anatomy, School of Medicine, Mazandaran University of Medical Sciences, Khazar Boulevard, Sari, Iran.

Abstract

In the present study, we examined the antioxidative activities of Satureja khuzestanica essential oil (SKE) and possible protective effect of SKE on lipid profile, atherogenic index and liver enzyme markers in Alloxan-induced Type 1 diabetic rats. Thirty male rats were randomly divided into three groups; group one as control, group two diabetic untreatment, and group three treatments with SKE by 500 ppm in drinking water, respectively. Diabetes was induced in the second and third groups by alloxan injection subcutaneously. After 8 weeks, the levels of fasting blood glucose (FBG), triglyceride (TG), cholesterol (C), low density lipoprotein (LDL), very low density lipoprotein (VLDL), high density lipoprotein (HDL), atherogenic index and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) of all groups were analyzed. Data were analyzed through non-parametric Man Whitney test (using SPSS 13 software) and p < 0.05 was considered significant. SKE inhibited significantly the activities of ALT and ALP and decrease FBG, TG, C, LDL and VLDL. HDL level was significantly increased when treated with the extract. The activities of AST stayed unaltered. Moreover, total antioxidant capacity of SKE was 3.20 ± 0.40 nmol of ascorbic acid equivalents/g SKE. This study showed that SKE is a source of potent antioxidants. The findings of the present study also suggest that SKE exert beneficial effects on the lipid profile, atherogenic index and liver enzymes activity in Alloxan-induced Type 1 diabetic rats.

Keywords


 

 Introduction

   Diabetes mellitus, one of the leading metabolic syndromes, accounts for highest morbidity and mortality worldwide (1). Diabetes mellitus is characterized by abnormalities in carbohydrate, lipid and protein metabolism due to complete or relative insufficiency of insulin secretion from pancreatic β-cells and/or defect in insulin action (2). Oxidative stress is a state of imbalance between oxidant and antioxidant systems (3).

   Recently, much attention has been focused on the central and key role of oxidative stress in the pathogenesis of different diabetic complications (4). Several studies have shown that antioxidant treatment reduces diabetic complications (5).

   Due to increasing demand of patients for the use of natural products and other herbal drugs with anti-diabetic activity, the general trend now is to use the natural products for medicinal application in their natural available form (6). Polyphenols, well-known antioxidants, have also proved to function as antidiabetic by reducing blood glucose levels (7-9). Researchers have become recently interested in investigation and research into extraction of natural antioxidants from medical herbs to replace synthetic antioxidants (10-13). Therefore, the research into the determination of the natural antioxidant source is very important to promote public health (12).

   Satureja-khuzestanica, an endemic plant of Iran, decreases glucose and malondealdehyde in serum diabetic patients (14-16). The components of this extract were analyzed with gas chromatography/mass spectrometry (GC/MS) in Research Center of Lorestan University as reported in our previous paper (16). The main component of this extract is carvacrol as a good antioxidant (14). Since the hypolipidemic, antiatherogenic and liver protective effects of SKE have not previously been reported; the objectives of the present study were to investigate hepatoprotective, hypolipdemic and antiatherogenic effects of Satureja khuzestanica essential oil in Alloxan-induced Type 1 diabetic rats.

Experimental

Isolation of the essential oil from Satureja khuzestanica

   Satureja khuzestanica essential oil was prepared from cultivated satureja khuzestanica in Khorramabad (Lorestan province, western Iran). The aerial parts of the plants were collected during flowering stage and were air-dried at ambient temperature in the shade. The aerial parts were hydro-distilled using a Clevenger apparatus for 4 h, giving yellow oil in 0.9% yield. The oil was dried over anhydrous sodium sulfate and stored at 4ºC. The plant was previously identified by the Department of Botany of the Research Institute of Forests and Rangelands (TARI) in Tehran, Iran. A voucher specimen (No. 58416) has been deposited at the TARI Herbarium (14, 16).

   The components of Satureja khozestanica essential oil were analyzed with gas chromatography/ mass spectrometry (GC/MS) in Research Center of Lorestan University (Table 1). The complete details of this GC/MS will be published in future.

 

Table 1. The components of Satureja khuzestanica essential oil that analyzed by GC/MS.
 

No.

Compound name

Area (%)

No

Compound name

Area (%)

1

3-Methyl butanol

0.14

19

β-Phellandrene

0.34

2

Eugenol

1.33

20

α -Thujene

1.26

3

1,8-Cineole

0.24

21

β -Caryophyllene

0.7

4

α -Pinene

0.99

22

γ-Terpinene

2.77

5

Geranyl acetone

0.5

23

Camphene

0.14

6

cis-Sabinene hydrate

0.68

24

α-Franesene

0.7

7

iso-Amylpropionate

0.23

25

Terpinolene

0.22

8

β-Bisabolene

3.77

26

β-Pinene

0.32

9

Linalool

3.32

27

α-Bisabolene

0.51

10

Myrcene

2.43

28

Nonanal

0.24

11

Caryophyllene oxide

1.53

29

trans-2-Carene-4-ol

0.73

12

4-Terpineol

4.1

30

β-Udesmol

0.32

13

iso-Butyl-2-methyl butyrate

0.19

31

α-Terpineol

0.42

14

Heptadecane

0.19

32

3-Carene

0.36

15

Thymyl methyl ether

1.21

33

α-Bisabolol

0.27

16

α-Terpinene

0.73

34

trans-Dihydrocarvone

0.26

17

Musk ambrette

0.08

35

para-Cymene

5.61

 

Animals

   Thirty male mature Sprague-Dawley rats (180-200 g) were obtained from Pasteur Institute of Tehran and were allowed to adapt themselves with the new location for one week. This study was approved by the Animal Ethics Committee of the Medical University of Lorestan with accordance to the national health and medical research council guidelines. The rats were divided to tree groups (10 per each). The studied groups were as follows: group 1 as control, group 2 as diabetic without treatment and the 3rd group as diabetic treatment with .SKE

Diabetes induction

   Diabetes was induced after overnight fasting in the second and third groups by injection of alloxan monohydrate (120 mg/Kg) subcutaneously (17). Beta-cell degradation by alloxan leads to release of more insulin. Owing to acute hypoglycemia, the rats received 10% sucrose solution for 48 h instead of drinking water. Five days after induction of diabetes, blood samples were gathered from the end part of tails. Blood glucose was measured by glucometer and the rats with blood glucose level of ≥ 300 mg/dL (16.7 mmol/L) were considered as diabetic (18). During the first five days after diabetes induction, 1-3 rats per group died because of alloxan toxicity. The rats were kept at 12/12 dark-light period in 21 ± 3°C temperature. All animals were allowed free access to food and water ad libitum during the experiment.

   The third group was treated with SKE by 500 ppm in drinking water, respectively for eight weeks (16). The treatment was begun at the first day of diabetes induction. After 8 weeks of treatment, animals were anesthetized (Nesdonal 50 mg/Kg, IP), blood samples were obtained from hearts and allowed to clot for 20 min in laboratory temperature and then centrifuged at 2000 rpm for 10 min for serum separation (14).

Biochemical study

   The serum levels of fasting blood glucose (FBG), triglyceride (TG), cholesterol (C), low density lipoprotein (LDL), very low density lipoprotein (VLDL), high density lipoprotein (HDL), atherogenic index and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) of all groups were analyzed.

   FBG, Cholesterol and triglyceride concentrations and ALT, AST and ALP activity were measured via biochemical analyzer using commercial kits (Olympus AU-600, Tokyo, Japan). HDL was measured in the supernatant after the precipitation of the Apo-B containing lipoproteins (LDL and VLDL) using polyanions in the presence of a divalent cation (18). LDL and VLDL were determined by calculation using the Freidewald equation (20, 21).

Total antioxidant activity

   Total antioxidant activity of the test samples was determinated according to the method of prrieto et al. In brief, 0.3 mL of the sample was mixed with 3.0 mL of reagent solution (0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate). Reaction mixture was incubated at 95ºC for 90 min under water bath. Absorbance of all the sample mixtures was measured at 695 nm. The total antioxidant activity was expressed as the number of equivalents of ascorbic acid acid (μmol g−1) (22).

Statistical analysis

    All values are expressed as mean ± SEM. The data were compared between groups by Mann-whitney U-test. Statistical analyses were performed using the SPSS 13 for windows software. A p-value < 0.05 was considered statistically significant.

Results and Discussion

Effect of SKE on serum lipid profile and atherogenic index

   Diabetes significantly increased serum FBG, TG, Cholesterol, VLDL and LDL concentrations in comparison with the control group. Treatment of diabetic animals with SKE significantly inhibited an increase inserum FBG, TG, Cholesterol, VLDL and LDL concentrations and atherogenic index in comparison with the untreated diabetic animals. The treatment of diabetic animals with SKE also significantly inhibited decrease of serum HDL concentrations in comparison with the untreated diabetic animals (p < 0.05) (Table 1). There are reports that several medicinal plant such as Cassia auriculata flowers (23), ginger rhizome (Zingiber officinale) (24) and fruits of Musa AAA (Chenkadali) (25) have hypolipidemic effects. What’s more, there are reports that natural antioxidant such as lycopene and Natural phenolic compounds have hypolipidemic effects (26-28). Therefore medicinal plant and natural antioxidant with hypolipidemic effects could prevent or be helpful in reducing the complications of lipid profile seen in diabetic patients. The mechanism of hypolipedemic and antiatherogenic action of medicinal plant may be due to the inhibition of dietary lipid absorption in the intestine or its production by liver or stimulation of the biliary secretion of cholesterol and cholesterol excretion in the faeces (29, 30). The mechanism of hypolipedemic and antiatherogenic action of medicinal plant may also be due to the inhibition of glycation lipoproteins, enzymes and proteins that involve lipid and lipoprotein metabolism (31).

Effect of SKE on serum ALT, AST and ALP activity

   Serum ALT, AST and ALP activity as markers of liver function significantly (p < 0.05) were increased in the untreated diabetic animals in comparison with the control group. Treatment of the diabetic animals with SKE could significantly inhibit an increase of serum ALT and ALP activity in comparison with the untreated diabetic animals. Treatment by SKE could maintain serum ALT and ALP activity of the treated animal at the same level as that of the control group (Figures 1 and 2). AST activity remained unaltered (Figure 3). ALT, AST and ALP are considered to be biochemical markers for assessing liver function (32, 33). Hepatotoxicity is evidenced by an elevation of the serum marker enzymes (33). There are reports indicating that several medicinal plants such asAgyanom mixture, Bolex bitters and Remedia mixture can reduce these liver enzymes markers (34, 35). There are also reports that natural antioxidant such as coenzyme Q10 and melatonin reduces these liver enzymes markers. Therefore medicinal plant and natural antioxidant with hepatoprotective action could prevent or be helpful in reducing the complications of hepatic damage seen in diabetes patients (36, 37).

  

Figure 1. The effects of 500 ppm Satureja khuzestanica essential oil on serum ALP in alloxan induced diabetic rats.
*Significant change in comparison with diabetic without treatment at p < 0.05.

 

 

Figure 2. The effects of 500 ppm Satureja khuzestanica essential oil on seruml ALT in alloxan induced diabetic rats. *Significant change in comparison with diabetic without treatment at p < 0.05.

 

 

 

Figure 3. The effects of 500 ppm Satureja khuzestanica essential oil on serum AST in alloxan induced diabetic rats.

 

Total antioxidant activity of SKE

   The phosphomolybdenum method has been widely used in the assessment of total antioxidant activity of plant extracts, natural compounds and foods. Figure 4 shows the total antioxidant activity of Ascorbic acid as standard. The total antioxidant activity of SKE was 3.20 ± 0.40 nmol of ascorbic acid equivalents/g SKE. The difference in the amount of antioxidant of extracts may be attributed to the differences in the amount and kind of existing antioxidant compounds in them such as carotenoids, phenol and ascorbic acid (38). The antioxidant activity has been shown by the SKE may be due to the presence of carvacrol, tannins, triterpenoids, steroids and flavoniods (16, 41). Carvacrol is agood antioxidant-scavengers of peroxyl radicals (42) and anti-inflammatory property (43).

 

 

Figure 4. Total antioxidant activity Free radical of Ascorbic acid as standard measured in phosphomolybdenum method. Each point represents the mean of five experiments.

 

 

Table 2. The effects of 500 ppm Satureja khuzestanica essential oil on TC, TG, LDL-C, HDL-C, VLDL-C and atherogenic index in diabetic rats.

Parameter

Control

Diabetic

Diabetic + SKE

FBG (mg/dL)

81 ± 28*

365 ± 64

287 ± 47*#

TG (mg/dL)

67.00 ± 16.66*

102.00 ± 25.01

72.00 ± 22.78*

TC (mg/dL)

71.00 ± 16.10*

118.00 ± 25.14

87.83 ± 24.14*#

HDL (mg/dL)

34.66 ± 8.90*

29.75 ± 10.90

33.54 ± 9.64*

LDL (mg/dL)

19.60 ± 3.87*

70.85 ± 12.24

38.43 ± 9.94*#

VLDL (mg/dL)

13.40 ± 3.33*

20.40 ± 5.00

14.40 ± 4.56*

Atherogenic index (units)

TC/HDL-C

2.05 ± 0.54*

3.97 ± 0.75

2.62 ± 0.62*

LDL/HDL-C

0.57 ± 0.08*

2.38 ± 0.67

1.15 ± 0.27*#

 Values are represented as mean ± SEM SKE: Satureja Khuzestanica essential oil.
*Significant change in comparison with diabetic without treatment at p < 0.05.
#Significant change in comparison with control at p < 0.05.

  

    Our recent results indicated that SKE is found to possess a good antioxidant activity. Researches reported the role of oxidative stress as a central factor in the onset and progression of diabetic complications such as hyperlipemia and hepatic damage (4, 39, 40). Numerous reports together with our results have proved the efficacy of antioxidative supplements administration in the prevention of diabetic complications. Antioxidant therapy is used as one of the most important treatment strategies for diabetic patients for the prevention and slowing of diabetic complications progression such as hyperlipemia, hepatic damage. Moreover; beneficial effects of SKE as antioxidant, antidiabetic, anti-inflammatory and anti-hyperlipidemia, toxicity and terato-genicity tests were also performed and confirmed plant’s safety (19); moreover, this extract can be produced in large amount and low cost. For the safety of SKE, a polyphenolic compound with antioxidant and anti-inflammatory properties, consumption of SKE is introduced to diabetic patients.

Conclusion

   This study showed that SKE is found to posses a good antioxidant activity and has beneficial effects in reducing the elevated serum lipid profile, atherogenic index and liver enzyme markers of STZ-induced-diabetic rats. Hence, attenuation of lipid profile, atherogenic index and liver enzyme markers can decrease the risk of cardiovascular death and hepatic damage in diabetic patients.

 

(1)    Najm W and Lie D. Herbals used for diabetes, obesity, and metabolic syndrome. Prim. Care (2010) 37: 237-54.

(2)    Lin Y and Sun Z. Current views on type 2 diabetes. J. Endocrinol. (2010) 204: 1-11.

(3)    Koksal M, Eren MA, Turan MN and Sabuncu T. The effects of atorvastatin and rosuvastatin on oxidative stress in diabetic patients. Eur. J. Intern. Med. (2011) 22: 249-53.

(4)    Giacco F and Brownlee M. Oxidative stress anddiabetic complications. Circ. Res. (2010) 107: 1058-70.

(5)    Golbidi S, Ebadi SA and Laher I. Antioxidants in the treatment of diabetes. Curr. Diabetes Rev. (2011) 7: 106-25.

(6)    Resmi CR, Venukumar MR and Latha MS. Antioxidant activity of Albizzia lebbeck (Linn.) Benth. in alloxan diabetic rats. Indian J. Physiol. Pharmacol. (2006) 50: 297-302.

(7)    Hamden K, Allouche N, Damak M and Elfeki A. Hypoglycemic and antioxidant effects of phenolic extracts and purified hydroxytyrosol from olive mill waste in-vitro and in rats. Chem. Biol. Interact. (2009) 180: 421-32.

(8)    Aslan M, Orhan DD, Orhan N, Sezik E and Yeşilada E. A study of antidiabetic and antioxidant effects of Helichrysum graveolens capitulums in streptozotocin-induced diabetic rats. J. Med. Food. (2007) 10: 396-400.

(9)    Kamalakkannan N and Prince PS. Antihyperglycaemic and antioxidant effect of rutin, a polyphenolic flavonoid, in streptozotocin-induced diabetic wistar rats. Basic Clin. Pharmacol. Toxicol. (2006) 98: 97-103.

(10)     Wojcik M, Burzynska-Pedziwiatr I and Wozniak LA. A review of natural and synthetic antioxidants important for health and longevity. Curr. Med. Chem. (2010) 17: 3262-88.

(11)       Zhang J, Yuan K, Zhou WL, Zhou J and Yang P. Studies on the active components and antioxidant activities of the extracts of Mimosa pudica Linn. from southern China. Pharmacogn. Mag. (2011) 7: 35-9.

(12)       Vanzani P, Rossetto M, De Marco V, Sacchetti LE, Paoletti MG and Rigo A. Wild mediterranean plants as traditional food: a valuable source of antioxidants. J. Food Sci. (2011) 76: C46-51.

(13)       Bonilla J, Atarés L, Chiralt A and Vargas M. Recent patents on the use of antioxidant agents in food. Recent. Pat. Food Nutr. Agric. (2011) 3: 123-32.

(14)       Abdollahi M, Salehnia A, Mortazavi SH, Ebrahimi M, Shafiee A, Fouladian F, Keshavarz K and Kazemi A. Antioxidant, antidiabetic, antihyperlipidemic, reproduction stimulatory properties and safety of essential oil of Satureja khuzestanica in rat in-vivo: a oxicopharmacological study. Med. Sci. Monit. (2003) 9: 331-335.

(15)       Gohari AR, Saeidnia S, Kiuchi F, Honda G. Trypanocidal activity of some endemic species of Satureja in Iran. Iranian J. Pharm. Res. (2004) Supple 2: 72-73.

(16)      Tavafi M, Ahmadvand H, Tamjidipoor A, Delfan B and Khalatbari AR. Satureja khuzestanica essential oil ameliorates progression of diabetic nephropathy in uninephrectomized diabetic rats. Tissue Cell (2011) 43: 45-51.

(17)       Fernandes NP, Lagishetty CV, Panda VS and Naik SR. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract. BMC Complement. Altern.Med. (2007) 24: 29.

(18)      Haidara MA, Mikhailidis DP, Rateb MA, Ahmed ZA, Yassin HZ, Ibrahim IM and Rashed LA. Evaluation of the effect of oxidative stress and vitamin E supplementation on renal function in rat with streptozotocin-induced type 1 diabetes. J. Diabet Complications (2009) 23: 130-6.

(19)      Warnick GR, Cheung MC and Albers JJ. Comparison of current methods for HDL quantification. Clin. Chem. (1979) 25: 596-601.

(20)      Ahmadvand H, Ani M and Moshtaghie AA. Changes in biochemical parameters related to lipid metabolism following titanium treatment in rat. Iranian J. Pharm. Therap. (2010) 9: 69-71.

(21)        Friedewald WT, Levy RI and Fredrickson DS. Estimation of the concentration of LDL-C in plasma without use of the preparative ultracentrifuge. Clin. Chem. (1972) 18: 499-502.

(22)        Prieto P, Pineda M, and Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal. Biochem. (1999) 269: 337-41.

(23)     Vijayaraj PS, Muthukumar K, Sabarirajan J and Nachiappan V. Evaluation of antihyperlipidemic activity of ethanolic extract of Cassia auriculata flowers. Indian J. Biochem. Biophys. (2011) 48: 54-8.

(24)      ElRokh el-SM, Yassin NA, El-Shenawy SM and Ibrahim BM. Antihypercholesterolaemic effect of ginger rhizome (Zingiber officinale) in rats. Inflammopharmacology (2010) 18: 309-15.

(25)     Kaimal S, Sujatha KS and George S. Hypolipidaemic and antioxidant effects of fruits of Musa (Chenkadali) in alloxan induced diabetic rats. Indian J. Exp. Biol. (2010) 48: 165-73.

(26)      Jiang F and Dusting GJ. Natural phenolic compounds as cardiovascular therapeutics: potential role of their antiinflammatory effects. Curr. Vasc. Pharmacol. (2003) 1: 135-56.

(27)       Bose KS and Agrawal BK. Effect of lycopene from tomatoes (cooked) on plasma antioxidant enzymes, lipid peroxidation rate and lipid profile in grade-I hypertension. Ann. Nutr. Metab. (2007) 51: 477-81.

(28)      Kaliora AC and Dedoussis GV. Natural antioxidant compounds in risk factors for CVD. Pharmacol. Res. (2007) 56: 99-109.

(29)       Garjani A, Fathiazad F, Zakheri A, Akbari NA, Azarmie Y, Fakhrjoo A, Andalib S and Maleki-Dizaji N. The effect of total extract of Securigera securidaca L. seeds on serum lipid profiles, antioxidant status, and vascular function in hypercholesterolemic rats. J. Ethnopharmacol. (2009) 126: 525-32.

(30)      Jayasooriya AP, Sakono M, Yukizaki C, Kawano M, Yamamoto K and Fukuda N. Effects of Momordica charantia powder on serum glucose levels and various lipid parameters in rats fed with cholesterol-free and cholesterol-enriched diets. J. Ethnopharmacol. (2000) 72: 331-6.

(31)      Harris CS, Beaulieu LP, Fraser MH, McIntyre KL, Owen PL, Martineau LC, Cuerrier A, Johns T, Haddad PS, Bennett SA and Arnason JT. Inhibition of advanced glycation end product formation by medicinal plant extracts correlates with phenolic metabolites and antioxidant activity. Planta Med. (2011) 77: 196-204.

(32)       Thattakudian Sheik Uduman MS, Sundarapandian R, Muthumanikkam A, Kalimuthu G, Parameswari SA, Vasanthi Srinivas TR and Karunakaran G. Protective effect of methanolic extract of Annona squamosa Linn in isoniazid-rifampicin induced hepatotoxicity in rats. Pak. J. Pharm. Sci. (2011) 2: 129-34.

(33)        Murat Bilgin H, Atmaca M, Deniz Obay B, Ozekinci S, Taşdemir E and Ketani A. Protective effects of coumarin and coumarin derivatives against carbon tetrachloride-induced acute hepatotoxicity in rats. Exp. Toxicol. Pathol. (2011) 63: 325-30.

(34)       Akande IS, Ebuehi OA, Samuel TA, Onubogu IC and Esin H. Effects of herbal remedies (Agyanom mixture, Bolex bitters and Remedia mixture) on hepatic and renal functions in male rats. Nig. Q. J. Hosp. Med. (2010) 20: 70-6.

(35)      Girish C, Koner BC, Jayanthi S, Rao KR, Rajesh B and Pradhan SC. Hepatoprotective activity of six polyherbal formulations in paracetamol induced liver toxicity in mice. Indian J. Med. Res. (2009) 129: 569-78.

(36)       Song HS, Kim HR, Park TW, Cho BJ, Choi MY, Kim CJ, Sohn UD and Sim SS. Antioxidant effect of CoQ(10) on n-nitrosodiethylamine-induced oxidative stress in mice. Korean J. Physiol. Pharmacol. (2009) 13: 321-6.

(37)      Gonciarz M, Gonciarz Z, Bielanski W, Mularczyk A, Konturek PC, Brzozowski T and Konturek SJ. The pilot study of 3-month course of melatonin treatment of patients with non-alcoholic steatohepatitis: effect onplasma levels of liver enzymes, lipids and melatonin. J. Physiol. Pharmacol. (2010) 61: 705-10.

(38)       Gorinstein S, Cvikrova M, Machackova I, Haruenkit R, Zachwieja Z and Katrich E. Characterization of antioxidant compounds in Jaffa sweeties and white grapefruits. Food Chem. (2004) 84: 503-10.

(39)       Cuerda C, Luengo LM, Valero MA, Vidal A, Burgos R, Calvo FL and Martínez C. Antioxidants and diabetes mellitus: review of the evidence. Nutr. Hosp. (2011) 26: 68-78.

(40)      Omar EA, Kam A, Alqahtani A and Li KM, Razmovski- Naumovski V, Nammi S, Chan K, Roufogalis BD and Li GQ. Herbal medicines and nutraceuticals for diabetic vascular complications: mechanisms of action and bioactive phytochemicals. Curr. Pharm. Des. (2010) 16: 3776-807.

(41)      Moghaddam FM, Farimani MM, Salahvarzi S and Amin G. Chemical constituents of dichloromethane extract of cultivated Satureja khuzistanica. Evid. Based Complement. Altern. Med. (2007) 4: 95-8. Amanlou M, Dadkhah F, Salehnia A and Farsam H. An anti-nociceptive effects of hydroalcoholic extract of Satureja khuzistanica Jamzad extract. J. Pharm. Pharmaceut. Sci. (2005) 8: 102-6.

(42)     Amanlou M, Dadkhah F, Salehnia A and Farsam H. An anti-nociceptive effects of hydroalcoholic extract of Satureja khuzistanica Jamzad extract. J. Pharm. Pharmaceut. Sci. (2005) 8: 102-6.

(43)       Aeschbach R, Löliger J, Scott BC, Murcia A, Butler J, Halliwell B and Aruoma OI. Antioxidant actions of thymol, carvacrol, 6-gingerol, zingerone and hydroxytyrosol. Food Chem. Toxicol. (1994) 32: 31-6.