SciELO - Scientific Electronic Library Online

 
vol.19 issue2Isolation and Probiotic Potential of Lactobacillus Salivarius and Pediococcus Pentosaceus in Specific Pathogen Free ChickensBreed and Sex Differences in the Gross Anatomy, Digesta pH and Histomorphology of the Gastrointestinal Tract of Gallus Gallus Domesticus author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

Related links

Share


Brazilian Journal of Poultry Science

Print version ISSN 1516-635XOn-line version ISSN 1806-9061

Rev. Bras. Cienc. Avic. vol.19 no.2 Campinas Apr./June 2017

http://dx.doi.org/10.1590/1806-9061-2016-0452 

Articles

The Review of Certain In Vivo Antioxidant Effects on Essential Oils of Origanum Minutiflorum O Schwarz-Ph Davis, Juniperus Excelsa Bieb.subsp. Excelsa and Histopathologic Changes

I GözeI 

ÖF GözeII 

I YelkovanIII 

ŞA ÇetinusIV 

H SayginV 

N ErcanVI 

IGöze Pharmacy, Çarşıbaşı Street, No 7, 5800

IICumhuriyet University Faculty of Medicine Department of Pathology

IIIDepartment of Biology

IVFaculty of Science Department of Biochemistry

VSivas Numune Hospital Department of Urology

VICumhuriyet University Faculty of Veterinary Medicine Department of Biochemistry Sivas

ABSTRACT

Essential oil of plants called Juniperus excelsa Bieb. (JE), Origanum minutiflorum O. Schwarz and P.H. Davis (OM) were used in this study. In order to determine experimental doses, LD50 values of essential oils were determined on mice. Taking into consideration the LD30 range, the experimental toxic doses were calculated for each rat (rat/kg). The toxic dosages thus determined were adapted to rats for active substances (rat/kg). Using commercially available pure virgin olive oil (VOO) as the solvent and diluting agent, OM oil (n=10), JE fruitoil (n=10), carvacrol (CRV) (n=10), VOO (n=10) and normal saline SF (n=8) were administered on the basis of 12 days into intraperitoneal (IP). Enzyme activities of Glucose-6-Phosphate dehydrogenase (G6PDH), malate dehydrogenase (MDH), Superoxide Dismutase (SOD), Glutathione-S-transferase (GST), Adenosine-deaminase (ADA) and Catalase were studied in isolates of kidney, brain and liver tissues. The data was statistically analyzed through Kruskal Wallis variance analysis. Elevated levels of GST and catalase have been found statistically important, as have both essential oil activities of OM and JE in the kidney tissue (p<0.005). All of the enzymes except the levels of ADA and SOD led to a statistically significant change in the brain and liver. There was sinusoidal hyperemia and capsular adhesion in the liver as histopathological were found to be statistically significant (p<0.005). It did not observe any important changes in the other organs. Findings were scored and analyzed by using x2(chi-square) test and Fisher’s definite variance analysis.

Keywords: Brain; histopathology; antioxidant enzymes; Juniperus excelsa; kidney,liver; Origanum minutiflorum

INTRODUCTION

There are about 70 kinds of Juniperus L. (Cupressaceae) in the world. The type of Juniperus is represented by 10 taxons with seven sub types in Turkey. Juniperus excelsa Bieb (gray tall juniper), (Cupressaceae) which is an evergreen and coniferous tree, covers 82% of juniper forests in Turkey. It has disseminated to all countries within Iranian-Turan climate zone. The fruits of this tree are registered as juniper berries in Turkish codex (Demirhan, 2011); and the researches conveyed show that it involves alpha-pinene as active ingredient in its compound (Adams et al., 2013).

The fruits of Juniperus excelsa Bieb (J.E.) are being used as diuretic, stimulant, antiseptic and for treatment of injuries by Anatolians since the ancient ages. It is known that, its extracts bear the effects of being anti-inflammatory, anti-microbial, insecticide, anti-termite, hypotensive, diuretic; and also used as antiseptic for urinal system (Adams 1990a; Adams 1990b; Adams 2011; Demirhan 2001; Adams et al., 2013). Also it is used as flavor or fragrance material in some foods and beverages, by distilling the wood and leafs of some kinds of juniper, the juniper essence which is produced is used in perfumery and medicine industry (Adams et al., 2013; Yesenofski 1996). In various studies activities of antioxidant, antibacterial, antispasmodic and chemical composition of JE were found (Dadalıoğlu & Evrendilek, 2004; Sokovic et al., 2004; Topçu et al., 2005; Asilli et al., 2008; Moein et al., 2010; Emami et al., 2011; Orhan et al., 2011; Taviano et al., 2011; Ataş et al., 2012; Ehsani et al., 2012; Khan et al., 2012; Orhan et al., 2012; Karapandzova et al., 2014; Moein, 2014). The type of Origanum is represented by 23 kinds 32 taxons in Turkey and 41 kinds 52 taxons in the world (Baser et al., 1993; Baser, 2002).

Thyme Origanum minutiflorum O. Schwarz and P.H. Davis (O.M.), which is known as ‘Dairy thyme’ and ‘Tote thyme’ in the thyme world market, is an endemic kind that has disseminated only at Dairy (Sütçüler) region in Isparta city and contains 80 % of carvacrol and 4.0 & p-cimen (Baser et al., 1993; Baser, 2002; Dadalıoğlu & Evrendilek, 2004; Goze et al,2010).

Some kinds of Origanum are also used as spice in food, and in medicine among people against indigestion, inappetency and cough. It bears the features such as being antiseptic, sedative, expectorant and cramp healing. It is used in cosmetics, alcoholic or alcohol free beverages. Due to its anti-bacterial effect on bacteria which causes food spoil and food poisoning, it is a most wanted medicinal plant. It is generally known for its rich essential oils, antioxidant, antibacterial, antiviral and antifungal activities (Baser et al., 1993; Davis et al., 1998; Baytop 1999; Baser, 2002; Dadalıoğlu & Evrendilek, 2004; Baydar, 2005; Baser, 2008; Goze et al., 2010; Oke 2010; Kılıçgün & Korkmaz 2014). Owing to this fact, for the natural active ingredients to be defined in plants, which are used for medication or food, it is important to convey researches that examine the biological activity, toxicological and pharmacological dose-response relation.

This study aims to determine the in vivo antioxidant activity of O.M. and J.E. on kidneys which are used orally and pursuit possible tissue changes histopathologically. Although there are many studies about in vitro antioxidant and antibacterial activities of active ingredients of O.M. and J.E., there are not many studies of in vivo. Therefore, the research findings are the first one in this field.

MATERIAL AND METHOD

JE fruit and OM were harvested from Isparta, Sütçüler flora. The samples of the plant were registered under JE (CUFH Voucher No. ED11003) and OM (CUFH Voucher No. ED 1101) numbers at the Herbarium of Cumhuriyet University Faculty of Science and Literature Department of Biology.

Plant samples which were dried in shade and pulverized, were held in Clevenger apparatus for approximately 3 hours being exposed to hydro distillation. The oil yield of 500gr dried plant was determined as 2.1 ml for JE and 10 ml for OM.

In the study, 40 healthy 180gr and approximately 4 weeks old adult albino Wistar rats (Rattus Norvegicusvar. Albino) were used; and in order to determine the LD50 dose, again, 40 healthily 25-30gr, 4 weeks old rats (MusMusculusvar. Albino) were used. The experimental animals were provided from Experimentary Animals Laboratory at Cumhuriyet University. Experimental Animals Ethical Committee’s approval was gained (approval date and no: 1/9/2005-116). The animals were preserved under standard laboratory circumstances, with free feed and water at room temperature (21 ± 3 ºC).

During literature searches no studies about OM and JE, LD50 doses were found. Therefore, LD50 dose of oils was determined by using rats. The toxic dose was calculated as rat/kg for each and found as LD30 (for JE -LD30:160 ul). The animals which were tried on with Pure OM and pure carvacrol died at all the levels of doses given. Thus, OM and carvacrol (CRV) were mixed into olive oil and a depot/store solution was prepared (1 OM+3 Olive oil); every rat was given 100u (LD30) i.p. from this solution and this dose was determined as LD30 for OM. In the same way, by applying 100ul to each rat from the store solution which was prepared by using carvacrol (1 carvacrol +6 olive oil), LD30 value was found for the carvacrol.

This experiment was made on rats. The rats were divided into 5 groups. To the 1st group of rats JE (160ul), to the 2nd group only olive oil (40 ul), to the 3rd group OM (100ul), to the 4th group carvacrol (100ul) were injected for 12 days with i.p. and 10 rats were used as control group (SF). At the end of 12 days the rats were killed with dislocation and their organs were taken into cooled boxes.

Preparation of Homogenates: Tissues were taken and washed with 0.15 KCl. These pieces of tissue were cut on a glass; kidney, brain and liver were homogenized by using rotating 1000 rpm/min via glass homogenizer (B. Braun) after added 0.15 M KCl ratio of 1:3(w/u) per 1 gram. The homogenates were centrifuged (Beckman Model J2) for 15 min at 4800 rpm. Glucose 6 phosphate dehydrogenase (G6PDH), (Beutler 1971); malate dehydrogenase (MDH), (Warburg & Christian 1931); Glutathione S-transferase (GST), (Habig et al., 1974); Superoxide dismutase (SOD), (Sun et al., 1988); Adenosine deaminase (ADA), (Giuisti, 1974); and catalase enzymes activities were studied in tissue as spectrophotometrically (Spectro UV-VIS Double Beam PC Scanning), (Aebi,1983). The data was statistically analyzed through Kruskal Wallis variance analysis by SPSS (Ver: 13.0) program. Standard error level is taken as 0.05 (Akgül, 2005).

The tissues samples which were sent to a pathology laboratory for routine follow up in automatic tracking device via handon Path Centre (Thermo Electron Corporation, UK). It observed adhesions among the intestine kidney, brain and liver of rats which were given JE in the macroscopic examination. After scored of findings analyzed with X2 test and precise analysis of variance Fisher (Akgül, 2005).

RESULTS

Results of kidney homogenates

The activities of GST and catalase were found important as a statistics for OM and JE (p<0.005). The difference between groups was not significant as G6PDH, MDH, ADA and SOD enzymes values compared with in pairs (p>0.05) Apart from these, there were significant difference in values (p<0.05). Between groups of OM and CRV, JE and CRV, JE and olive found significant differences to pairwise comparisons are made in terms of value in GST (p<0.005).

When catalase activity compared in pairs of groups OM and CRV, OM and VOO, JE and CRV, JE and olive oil values difference was found statistically significant as shown in Table1 and Table 2.

Table 1 Analyzes of enzymes activities in kidney homogenates 

G6PDH GST MDH ADA SOD CATALASE
OM 0.000165±0.0001 0.00150±0.0012 0.00023±0.0003 0.00047±0.0003 97.80±0.57 97.58±32.02
JE 0.00043±0.0003 0.00027±0.0002 0.00021±0.0001 0.00037±0.0000 97.96±0.84 117.95±21.80
CRV 0.00064±0.0004 0.00308±0.0019 0.00023±0.0001 0.00035±0.0001 98.13±0.45 117.75±34.76
VOO 0.00040±0.000 0.00230±0.0004 0.00022±0.0001 0.0027±0.0010 97.77±0.26 171.51±17.44
SF 0.00125±0.00012 0.00136±0.0001 0.00024±0.0001 0.00027±0.0001 98±0.60 129.48±32.87
Kw 10.19 22.69 4.35 4.44 3.34 18.20
p Value 0.037 0.000 0.003 0.366 0.502 0.001

GP6DH U/g pro; GST U/mg pro; MDH U/mg pro; ADA U/mg pro; SOD U/g pro; CATALASE U/mg pro

Table 2 Analyzes of enzymes activities in brain homogenates 

G6PDH GST MDH ADA SOD CATALASE
OM 0.00124±0.0012 0.00167±0.0015 0.0001±0.0001 0.00038±0.0007 97.45±0.56 2.07±0.75
JE 0.00032±0.0002 0.00096±0.0004 0.00025±0.0001 0.00017±0.0001 97.52±0.58 2.79±0.81
CRV 0.00088±0.0002 0.00470±0.0050 0.00013±0.0001 0.00005±0.0001 97.13±0.15 2.62±0.75
VOO 0.0019±0.0004 0.0027±0.0002 0.00014±0.0001 0.00006±0.0000 97.67±0.67 9.76±0.60
SF 0.00151±0.0003 0.00276±0.0003 0.00016±0.000 0.00026±0.0002 96.10±2.60 10.38±0.44
Kw 19.21 18.270 17.01 11.73 5.81 14.89
p Value 0.001 0.001 0.002 0.009 0.213 0.005

GP6DH U/g pro; GST U/mg pro; MDH U/mg pro; ADA U/mg pro; SOD U/g pro; CATALASE U/mg pro

Results of brain homogenates

The level of G6PDH between groups of VOO /JE and JE/SF (p<0.005); the level of GST between groups of JE-CRV (p<0.005); the level of MDH between groups of OM-JE and CRV-JE (p<0.005); the level of catalase between groups of OM- VOO and JE-VOO-SF, CRV-JE and SF were found important if statistically compared in pairs (Table 2).

Results of liver homogenates

The activity of G6PDH in between groups of OM-SF and JE-VOO and CRV/VOO/SF; the activity of GST between groups of JE-CRV and JE-VOO and JE-SF; the activity of MDH between groups of OM and VOO, the activity of catalase between groups of OM/VOO and SF were found statistically important.

The enzymes of G6PDH, MDH, ADA and SOD did not show significant differences between the groups compared in pairs (p>0.005) (Table3).

Table 3 Analyzes of enzymes activities in liver homogenates  

G6PDH GST MDH ADA SOD CATALASE
OM 0.00099±0.00016 0.0017±0.0016 0.00032±0.00025 0.00006±0.0001 90.81±4.44 116.17±10.2
JE 0.0021±0.0015 0.00335±0.0023 0.00024±0.0001 0.0001±0.0001 93.52±0.89 98.13±8.49
CRV 0.0034±0.0004 0.00020±0.0001 0.00016±0.000 0.0001±0.00001 93.11±1.72 103±20.4
VOO 0.0039±0.0025 0.00021±0.0003 0.00001±0.000 0.00001±0.0000 92.61±2.68 90.21±2.465
SF 0.00448±0.001 0.0001±0000 0.00014±0.0011 0.00017±0.0002 91.83±2.07 90±3.368
Kw 18.87 20.12 12.89 19.94 4.32 14.89
p Value 0.000 0.000 0.012 0.364 0.364 0.005

GP6DH U/g pro; GST U/mg pro; MDH U/mg pro; ADA U/mg pro; SOD U/g pro; CATALASE U/mg pro

In histopathological examination, a finding was not seen in kidney tubule epithelia cell except cloudy swelling (p>0.05). After scored of findings analyzed with X2 test and precise analysis of variance Fisher. Fuzzy swelling (FS) in surface of tissue sample according to where tissue is turned off as semi quantitative (%), focal necrosis (FN) and the group of cell necrosis as a number (present or absent), Kupffer cell hyperplasia (present or absent), periportal inflammation (present or absent), sinusoidal hyperemia (SHR) (present or absent) were evaluated (Table 4). The sinusoidal hyperemia (SH) and capsular adhesion (CA) in liver were found statistically important (p<0.05) (Figure1). There are not significant changes which in other organs were observed.

Table 4 Evaluation of tissue samples in statistically 

GROUPS OM JE RESULTS
S % s %
SCN 0 11 68.8 3 100 P=0.530
1 5 31.3 - -
KOL 0 15 93.8 3 100 P=1.00
1 1 6.3 - -
PPI 0 2 12.5 - - X2=3.20 P=0.361
1 7 43.8 3 100
2 5 31.3 - -
3 2 12.5 - -
CA 0 7 43.8 2 66.7 X2=6.69 P=0.025
1 9 56.0 7 33.3
FS 0 5 31.3 3 100 X2=4.89 P=0.179
1 1 6.3 - -
2 2 12.5 - -
3 8 50.0 - -
SH 0 10 62.5 2 66.6 X2=7.71 P=0.021
1 6 37.5 1 33.3
2 -

SCN (Single Cell Necrosis), PPI (Periportal inflammation), KOL (Kollaps), FS (Fuzzy swelling), SH (Sinusoidal hyperemia), CA (Capsular Adhesion)

Figure 1 A general view of adhesions in the liver capsule (HEx40). 

Figure 2 The breakdown in the liver capsule adhesions: fibroadipose hipogralunom to similar tissue infiltration (HEx100). 

DISCUSSION

Serum enzyme activities are widely used in determining of organ functions in mammals. Also these are very important for environmental biology, searching for the effects of environmental pollutants on living organisms, the toxic effects of orally consumed plants and for bioavailability in recent years. These tests have been of great importance in recent years, particularly in the context of bioavailability, to explore the aquatic effects of environmental biology and environmental pollutants and the toxic effects of orally consumed plants.

The changes that may occur in the activities of these important enzymes working in the energy metabolic pathways may affect the formation of energy by the organism and affect the functioning of other metabolic pathways.

Superoxide dismutase (SOD), catalase (CAT), Glutathione reductase (GST) and malate dehydrogenase (MDH) are enzymatic compounds. Superoxide dismutase (EC 1.15.1.1, EC-SOD), as a superoxide anion is a form that part of the cellular antioxidant defense system by catalyzing the conversion in hydrogen peroxide to molecular oxygen. It plays an important role in controlling cellular superoxide levels in the compartment. It consists in high concentrations in the brain, liver, heart, kidney and erythrocytes (Fridovich, 1989).

In this study, SOD was found stable in all of the tissues. The mature erythrocytes don’t include ribosome, mitochondria and nucleus. Enzymes of SOD, catalase, GSH-Px and pentose phosphate pathway protected against O2 radicals. It can consider that OM and JE don’t cause inhibition of the metabolic pathway.

Glucose-6-phosphate dehydrogenase (G-6-PD) (D-glucose-6-phosphate: NADP+ 1-oxidoreductase, E.C. 1.1.1.49) is the first and control enzyme of the pentose phosphate pathway.G6PHD; it is an enzyme that plays a key role in the pentose, phosphate and glycolysis (Scott 1975). Also it has an anti-oxidant activity. G6PDH enzyme activity was found to decrease in homogenates of all tissues. According to these results, it can be observed that essential oils may generate inhibiting change.

Adenosine deaminase is purine metabolism enzyme which is responsible for immune mechanism (Wilson et al., 1991). Significant changes weren’t observed in ADA activity in all organs except for the brain. In this case, the tested substances aren’t effective in purine metabolism and suggest no creating damage.

MDH; is taken office in the maintenance of cell membrane integrity and the regulation of enzyme activity (Musrati et al., 1998). In this research, the levels of MDH was high in the liver and in the brain but was stabile in the kidney.

GST, glutathione plays an important role against sourced interior and exterior toxic chemicals in the cellular defense system (Armstrong, 1990). Reversible damage may have been created during the period of JE and OM application by various mechanisms in all tissues. In this case, it can be considered to change their enzyme activations to maintain the redox state of cells and especially repairing damage membranes by keeping to a minimum damage. It is very important for all of tissues.

Catalase (H2O2: H2O2 oxidoreductase, EC 1.11.1.6) has 4 units with a group hemoprotein and localized in intracellular organelles as peroxisomes. Its task is to destroy hydrogen peroxide to oxygen and water (Kono & Fridovich, 1982). This is quite low compared to the catalase activity in the kidneys of the control group that can cause essential oils erythrocyte catalase activity by inhibiting the accumulation of H2O2 and consequently hemolysis which is very important for all of tissues.

As a result, the activity of catalase was very meaningful in all tissues while the level of SOD was stable. The levels of GST and catalase were in kidney, ADA and SOD were important in liver. Statistical differences were found in all enzymes except for SOD. It can be observed that these types of plants consumed as a food of toxic effects should have more comprehensive studies. Additionally, the LD value for each study which uses the plant´s essential oils, need to be determinate. Because the composition of the plant essential oils vary depending on many reasons such as seasonal, droughts and temperature difference.

ACKNOWLEDGE

This study was supported by The Scientific Research Project Fund of Cumhuriyet University under project number SHMYO-5.

REFERENCES

Adams RP. The volatile leaf oils of Juniperus excelsa M-Bieb. From a native stand in Greece versus J. Excelsa cultivated at Kew, London. Journal Essential of Oil Research 1990a;2:45-48. [ Links ]

Adams RP. Juniperus procera of east Africa:volatile leaf oil composition and putative relationship to J. excelsa. Biochemical Systematic and Ecology 1990b;18:207-210. [ Links ]

Adams RP. Junipers of the world, the genus Juniperus. 3rd ed. Bloomington: Trafford Publishing, 2011. p. 428. [ Links ]

Adams RP, Tashev AN, Baser KHK, Christou AK. Geographic variation in volatile leaf oils of Juniperus excelsa M. Bieb. Phytologia 2013;95(4):279-285. [ Links ]

Aebi HE. Catalase. In: Bergmeyer UH, Bergmeyer J., Grassl M. Method of enzymatic analysis. Michigan: Verlag Chemie; 1983;3: 273-286. [ Links ]

Akgül A. Biyoistatistik. 3. baski. Ankara: Emek Ofset; 2005. [ Links ]

Armstrong RN. Glutathione s-transferases:reaction mechanism, structure and function. Chemical Research in Toxicology 1990;4(2):131-140. [ Links ]

Asili, J, Emami SA, Rahimizadeh M, Fazly-Bazzaz BS, Hassanzadeh MK. Chemical and antimicrobial studies of Juniperus excelsa subsp. Excelsa and Juniperus excelsa subsp. Polycarpos essential oils. Journal of Essential Oil Bearing Plants 2008;11(3):292-302. [ Links ]

Atas AD, Goze I, Alim A, Cetinus SA, Durmus N, Vural N, et al. chemical composition, antioxidant, antimicrobial and antispasmodic activities of the essential oil of Juniperus excelsa subsp. excelsa. Journal of Essential Oil Bearing Plants 2012;15(3):476-483. [ Links ]

Baser KHC. The turkish origanum species. In: Kintzios SE, editor. Oregano. The genera origanumand lippia. London: Taylor and Francais; 2002. p.108-126 [ Links ]

Baser KH. Biological and pharmacological activities of carvacrol and carvacrol bearing essential oils. Current Pharmaceutical Design 2008;14:3106-3119. [ Links ]

Baser KHC, Özek T, Tümen G, Sezik E. Composition of the essential oils of Turkish Origanum Specie swith Commercial Importance. Journal Essential of Oil Research 1993;5(6):619-23. [ Links ]

Baydar H. The effects of different harvest dates on essential oil content and essential oil composition in Origanum minutiflorum Schwarz. Akdeniz Üniversitesi Ziraat Fakültesi Dergisi 2005;18:175-178. [ Links ]

Baytop T. Therapy with medical plants in Turkey, past and present). Istanbul: Nobel Tip Kitapevleri; 1999. [ Links ]

Beutler E. Red cell metabolism manual of biochemical methods. London: Academic Press; 1971. [ Links ]

Dadalioglu I, Evrendilek GA. Chemical compositions and antibacterial effects of essential oils of Turkish oregano (Origanumminutiflorum), bay laurel (Laurusnobilis), Spanish lavender (Lavandula stoechasL), and fennel (Foeniculum vulgare) on common food borne pathogens. Journal of Agricultural and Food Chemistry 2004;29/52(26):8255-8260. [ Links ]

Davis PH, Mil RR, Tan K. Flora of Turkey land the east Aegean islands. Edinbourgh: Universty Press; 1998. v.10, suppl 1. [ Links ]

Demirhan Erdemir A. Sifali bitkiler dogal ilaçlarla gelen tedaviler. Istanbul: Alfa Basim Yayim Dagitim; 2001. [ Links ]

Ehsani E, Akbari K, Teimouri M, Khadem A. Chemical composition and antibacterial activity of two Juniperus species essential oils. African Journal Microbiology Research 2012;6(38):6704-10. [ Links ]

Emami SA, Abedindo BF, Hassanzadeh-Khayyat M. Antioxidant activity of the essential oils of different parts of Juniperus excelsa M. Bieb. subsp. Excelsa and J. excelsa M. Bieb. subsp. polycarpos (K. Koch) Takhtajan (Cupressaceae). Iranian Journal of Pharmaceutical Research 2011;10(4):799-810. [ Links ]

Fridovich I. Superoxide dismutases. An adaptation to a paramagnetic gas. Journal of Biological Chemistry 1989;264:7761-7764. [ Links ]

Goze I, Cetin A, Goze A Investigation of effects of essential oils of Origanum minutiflorum O Schwarz pH Davisand Cyclotrichium niveum (Labiatae) plants on angiogenesis in shell-less chick embryo culture. African Journal of Biotechnology 2010;9(14):2156-2160. [ Links ]

Giuisti G. Enzyme activities. In: Bergmeyer U, ed. Methods of Enzymatic Analysis. Gmblt Weinheim, Bergest: Verlog Chemie;1974;1092-1098. [ Links ]

Habig WA, Tabst MS, Jacoby WV. Glutatyon S transferase Biolchem. 1974; 7130-7139. [ Links ]

Karapandzova M, Stefkov G , Cvetkovikj I , Sela F , Panovska TK , Kulevanova S. Chemical characterization and radical scavenging activity of leaves of Juniperus foetidisima, J. Excelsa and J. Communis from Macedonian flora. Macedonian Pharmaceutical Bulletin 2014;60(2):29-37. [ Links ]

Khan M, Khan A, Rehman N, Gilani A.H. Pharmacological explanation for the medicinal use of Juniperus excelsa in hepatoprotective gastrointestinal and respiratory disorders. Journal Natural of Medicines 2012;66:292-301. [ Links ]

Kiliçgün H, Korkmaz M. Hepatoprotective and antidiabetic activity of origanum minutiflorum grown wild in Turkey. Bothalia Journal 2014;44:3. [ Links ]

Kono Y, Fridovich I. Superoxide radicals inhibit catalase. Journal of Biological Chemistry 1982;257:5751-5754. [ Links ]

Moein MR, Ghasemi Y, Moein S, Nejati M. Analysis of antimicrobial, antifungal and antioxidant activities of Juniperus excelsa M. B. subsp. polycarpos (K. Koch) Taghtajan essential oil. Pharmacognosy Research 2010;2(3):128-31. [ Links ]

Moein MR, Moein S, Mousavi F. Study on relationship between antioxidant potential and phenolic contents of Juniperus excelsa fruit. International Journal of Pharmacy and Pharmaceutical Science 2014;6(7):192-194. [ Links ]

Musrati RA, Kollárová M, Mernik N, Mikulásová D. "Malate dehydrogenase:distribution, function and properties". General Physiology and Biophysics 1998;17(3):193-210. [ Links ]

Oke F. Biological potentials and cytotoxicity of various extracts from endemic Origanum minutiflorum O. Schwarz & P.H. Davis. Food and Chemical Toxicology 2010;48:1728-1733. [ Links ]

Orhan N, Akkol E, Ergun F. Evaluation of anti inflammatory and antinociceptive effects of some Juniperus species growing in Turkey. Turkish Journal of Biology 2012;36:719-726. [ Links ]

Orhan N, Orhan, IE, Ergun F. Insights into cholinesteras einhibitory and antioxidant activities of five Juniperus species. Food and Chemical Toxicology 2011;49(9):2305-2312. [ Links ]

Scott WA. Glucose-6-phosphate dehydrogenase from neurospora crassa. Methods Enzymology 1975;41:177-182. [ Links ]

Sokovic MD, Ristic M, Grubisic D. Chemical composition and antifungal activity of the essential oil from Juniperus excelsa berries. Pharmaceutical Biology 2004;42:328-331. [ Links ]

Sun YI, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clinical Chemistry 1988;34(3):497-500. [ Links ]

Taviano MF, Marino A, Trovato A, Bellinghieri V, La Barbera TM, Guvenc A, et al. Antioxidant and antimicrobial activities of branches extracts of five Juniperus species from Turkey. Pharmaceutical Biology 2011;49:1014-102 [ Links ]

Topçu G, Gören AC, Bilsel, G, Bilsel M, Çakmak O, Schilling J, et al. Cytotoxic Activity and Essential Oil Composition of Leaves and Berries of Juniperus excelsa. Pharmaceutical Biology 2005;43(2):125-8. [ Links ]

Warburg O, Christian H (1931a). Aktivierung von Kohlehydrat in roten Blutzellen. Biochem. Z. 238:131. [ Links ]

Wilson DK, Rudolph FB, Quiocho FA. Atomic structure of adenosine deaminase complexed with a transition-state analog: understanding catalysis and immunodeficiency mutations. Science 1991;252(5010):1278-1284. [ Links ]

Yesenofski J. Juniper oil distillation and marketing project: western juniper commercialization program. Oregon: The Confederated Tribes of the Warm Springs Reservation; 1996. (Final Report). [ Links ]

Received: December 2016; Accepted: February 2017

Corresponding author e-mail address Nazlı Ercan Cumhuriyet University Faculty of Veterinary Medicine Department of Biochemistry Sivas, Turkey. 58140 Tel: (90-346) 2191010-2575 Email:nazliercan@yahoo.com

Creative Commons License This is an open-access article distributed under the terms of the Creative Commons Attribution License