Effect of Some Medicinal Plants: Urticadioica, Viscum album, Eucalyptus globulus and
Olea Europeaonalloxan-İnduced Diabetic Mice
Yusuf OZAY1 Muhammet Kasım CAYCI2 Sevda GUZEL3 Ebru Gökalp OZKORKMAZ4 Vahdettin BAYAZIT2 1Adiyaman University, Faculty of Medicine, Department of Medical Biology, 02200 Adiyaman, Turkey
2Dumlupinar University, Faculty of Arts and Sciences, Department of Biology, 43100, Kutahya, Turkey 3 Mersin University, Faculty of Pharmacy, Department of Pharmacognosy, 33169, Mersin, Turkey 4Yildirim Beyazit University, Institute of Health Sciences, 06690, Ankara, Turkey
Running Title:Effects of some medicinal plants on Diabetes
*Corresponding author: Assist. Prof.Dr. Yusuf OZAY Received: December 20, 2015 E-mail:yusufozay33@hotmail.com Accepted: February 02, 2016 yozay@adiyaman.edu.tr
Abstract
Current antidiabetic treatmentsare based on synthetic agents which existingseveral adverse effects in long term uses. Therefore, it is needed to develop new and safer pharmaceuticals as alternative for treatment of diabetes. Traditionally usedplants are important source of potential antidiabetic agents. In this study four plants used traditionallyfor treatment of many disease were evaluated for their antidiabetic action.This study was focused onthe effects of Urticadioica(UD),Viscum album(VA), Eucalyptus globulus(EG) andOleaeuropea(OE)extracts on blood and organ glucose concentrations of hyperglycemic mice. Hyperglycemia was induced by alloxan and the antidiabetic drug glutril used for the com-parison. Aqueous extracts of UD, VA,EG and OE andglutril were given three drops orally every day and blood glucose levels were measured in two days interval at eight periods.It was found that blood glucose concentrations of UD, VA, EG, OE and glutril groups were 90.9 3.9 mg/ dl, 95.22.4 mg/dl, 75.3±3.1 mg/dl, 93.7±2.7 mg/dl and 96.32.3 mg/dl, respectively (P<0.001). The all tested plants were decreased fasting blood glucose levels similar to antidiabetic agent glutril. Additionally, no adverse effects were determined in plant extracts treated groups. Fur-ther investigation is needed to determine constituents which is responsible from activity andthese plants can be use as alternative Fur-therapeutics.
Keywords:Hyperglycemia, Urticadioica, Viscum album, Eucalyptus globulus, Olea europea, Traditionalmedicine
INTRODUCTION
Diabetes mellitus (DM) leads to cause death and dis-ability worldwide with an increasing prevalence[1] is a pro-gressive metabolic disease of endocrine gland system and mainly characterized byhyperglycemia and lipidemia[2-4]. The illness is associated with serious structural and function-al complications like as retinopathy, neuropathy[3,5],renfunction-al failure, skin complications[6],stroke and peripheral vascular diseases[2] by affecting cells, tissues and organs[7].
In addition to insulin, various therapeutic agents are used for treatment of diabetes[8], but in long term uses these agents cause harmful effects such as severe hypoglycemia, lactic acidosis, peripheral edema[2], drug-resistance and weight gain[9]. Therefore, new approaches is needed to improve more efficient and safer agents for management of diabetes[6,7].Plants are rich source of potential new thera-peutic agents for alternative and complementary treatment of various disease including diabetes[6,8].Besides, herbal medicines which are cheaper and more effective than syn-thetic agents have been suggested for treatment of diabetes by World Health Organization (WHO)[9].Medicinal poten-tial of plants have been taken attention by depending on their traditional uses in different parts of the world[5,10]. Ethnobotanical research indicated that several plants have been traditionally used to control blood glucose levels and complications of diabetes since ancient times[5,6]. Previous studies indicated that several plants in different family inves-tigated for their antidiabetic activities[4, 7, 9, 10, 11].
In this study four plants (UrticadioicaL. (Urticaceae),Viscum album L.(Loranthaceae), Eucalyptus globulusLabill (Myrtaceae)and Olea europea L. (Oleaceae)) which traditionally used for treatment of various illness in
folk medicine were selected to investigate for their antidia-betic actions. Perennial herb U. dioica known as stinging nettle[12]traditionally used for treatment of many diseases including allergic rhinitis, cardiovascular diseases, hemor-rhoids[13], rheumatism, eczema,anaemia[14],stomachac he[15] and diabetes[16]. The plant contained various types of secondary metabolites such asphenolics, flavonoids, phenylpropanoids, lignans[14],coumarins, sterols, terpe-noids and lec¬tins[17]. Literature data indicated that the plant has several biological activities such as anti-inflamma-tory [14,18],antioxidant[17,18],immunostimulation, antian-algesic and antimicrobial[18]. Half parasitic plant V. album called as mistletoe is distributed throughout the world[19] and traditionally used for the treatment of fever, asthma[20], epilepsy, headache[21],diabetes, cancer and hyperten-sion[22]. Phytochemical studies indicated that the plant contained polysaccharides, phenylpropanes, lignans, flavo-noids, lectins, viscotoxins[19,21],terpeflavo-noids, alkaloids[20], phenols and amines[19]. There are some reports on biologi-cal activities of V. album including antiinflammatory, anti-nociceptive[19,21], anticancer, immunomodulatory[20,23], antioxidant and antimicrobial activities[19]. Evergreen tree E. globulus, which commonly known as Tasmanian Blue Gum[24], contained essential oil, flavonoids, tannin[25], terpenoids[26] and traditionally used as treatment of cold, influenza, diarrhea, toothache25] and diabetes[27]. Previous research showed that the plant had antioxidant, antimicro-bial[24,28], antiinflammatory, anthelmintic[28] and insec-ticidal activities[24].Well known evergreen tree O. europea named as Olive[29] were distributed throughout Mediterra-nean region[29,30]and traditionally used in treating fever, malaria[31] and diabetes[32]. Phytochemical investigations
showed the presence of phenolics[29-33], flavonoids[29] vi-tamins and polyphenols[33]. Also antibacterial[29], antioxi-dant[30] and anti-inflammatory [31] activities of the plant have been reported.
In the present work four medicinal plants (U. dioica(UD), V. album (VA), E. globulus (EG) and O. europea (OE)) which arecommon inTurkey were selected to studyfor their antidiabetic activities.The aim of the study was to investi-gate the blood glucose concentrations, body weights, organ glucose levels and weights of liver, brain, kidney and brain withhealthy and alloxan induced diabetic male mice after treatment of aqueous extracts obtained from the plants for 15 days.
MATERIALS and METHODS
Plant material
Plant materials were collected from two different re-gions of Turkey during the summer of 2003 and identified by one of the authors (Y. Ozay). Half parasitic plant V. album samples were collected from host plant Salix alba. The plant materials, their families andorigins were listed in Table 1.
Chemicals
Alloxan monohydrate was purchased from Sigma Chem-ical Company and glutril was purchased fromRoche. Dis-tilled water was used for the preparation of all the samples and solutions. Freshly prepared solutions were used. Glutril including glibornuride was used as oral antidiabetic agent for positive control and prepared as one thumbnailstablet(25mg) added1ml of distilled water.
Experimental animals
The experimental animals obtained from the central animal house of Dumlupinar University were male mice (Mus musculus) at the age of 2 months, with body weight about 25 g. The animals were randomly divided into ten groups of ten mice each. All the experiments wereperformed with the acceptance ofethical commission of Dumlupinar University,Facultyof Arts and Sciences, Kutahya, Turkey. The animals were kept under standard conditions (12 h light/ 12 h dark cycle; 25°C ± 3°C and 50–70 % humidity) and fed with standard pellet diet (17% crude protein) ad libitum with unlimited access to water.
Various treatment groups were designed. Group I: un-treated control group, Group II, Group III, Group IV, Group V were plant extract treated groups (UD, VA, EG, OE treat-ed groups, respectively). Group VI: glutril treattreat-ed group and Group VII, Group VIII, Group IX, Group X were plant extract treated groups (UD, VA, EG, OE treated groups, re-spectively) with hyperglycemia induced by alloxan. Finally, groups I to V were non-diabetics and groups VI to X were diabetic.
Experimental induction of diabetes
Animals were administered intraperitoneally (i.p.) with a freshly prepared solution of alloxan monohydrate in
sa-line at a dose of 150 mg/kg of bodyweight and after 20 days blood glucose levels were measured from tail vein by glu-cometer (Roche), levels of 200 mg/dl or greater were ac-cepted as diabetic and chosen for the experiments.
Extraction procedure
Air dried and powdered leaves (50 g) of UD, VA, EG and OE were extracted with distilled water. Then all suspen-sions were separately centrifuged for 5 min at 5000 rpm and liquid parts were taken to give aqueous extracts of studied materials. The all extracts were stored in dark at 4 °C until further use.
Determination of blood glucose levels
The aqueous extracts of the plants and glutrilwere ad-ministered 3 drops (0.1 ml, 100 mg/kg) orally with full stomachevery day and blood glucose concentrations were measured in two days interval at eight periods. After 15 days blood glucose levels of both diabetic and non-diabetic groups were monitored by glucometer readings. Addition-ally in the beginning and at the end of experiments animals were weighed in order to compare the effects of extracts on animal weight (Table 2).
Determination ofglucose levels in liver,brain,kidney andheartand organ weights
After 15 days treatment of each group, brain, heart,liver and kidneysamples of animals were taken for further studies. Due to brain, heart,liver and kidneysampleswere dissectedan-dremovedafter being killedbycervicaldislocation thenorgans were weighed. Tissues were added distilled water, homog-enized andcentrifuged for 6 min at 5000 rpmthenwarmed to 37 °C for 5 min. Organ glucose concentrationswere mea-sured with glucose determination kit (LiquickCor-Glucose, Cormay, Poland) at 671 nm by spectrophotometer (Milton Ray com. speet. 20D).The measurement resultswere applied as described in Cormay and sampleglucose concentration-swere calculated[34].
Statistical analysis
The experimental data were expressed as mean ± SD and statistical analyses wereevaluated using SPSSversion 17.0 and Mann-Whitney U test. <0.05 was considered statisti-callysignificant.
TABLE 1: LIST OF PLANT MATERIALS, FAMILIES AND THEIR ORIGINS
Plant material Family Locality
Urticadioica Urticaceae Bolcekvillage,Kutahya,Turkey Viscum album Loranthaceae Tavsanli, Kutahya, Turkey Eucalyptus globulus Myrtaceae Karacailyas, Mersin, Turkey
RESULTS
Determination of blood glucose levels
In this study aqueous extracts of UD, VA, EG and OE were tested for their antidiabetic actions and glutril used as positive control. Theresults of blood glucose levels and body weightsof both diabetic and non-diabetic groups were listedin Table 2. In non-diabetic groups,last measurements of blood glucose levels of UD, VA, EG, OE treated and con-trol groups were in normal value with the range of 93.2±2.4-67.5±3.5 mg/dl. All tested plant extracts were kept blood glucose levels in normal values (Table 2). When compared with the diabetic groups between each other it was found that UD, VA, EG, OE and glutriltreated groups were significantly reducedblood glucose concentra-tions to90.93.9 mg/dl, 95.22.4 mg/dl, 75.3±3.1 mg/dl, 93.7±2.7 mg/dl and 96.32.3 mg/dl, respectively (P<0.001). Aqueous extract of EG was found more effectiveness than UD, VA OE and glutril. Blood glucose level of EG treated group was significantly reduced from372.2±12.3 to 75.3±3.1 while blood glucose levels of glutril treated group were reduced from 374.6±5.8to 96.3±2.3(P<0.001). Addition-ally initial and finalmeasurements of body weight indicated that there were no significant difference>0.05 observe-din both tested diabetic and non-diabetic groups (Table 2).
Determination ofglucose levels in liver,brain,kidney andheartand organ weights
In this work we weighed organs which used to determine organ glucose levels to compare effects of plants extracts on all tested groups. Due to liver, brain, heartand kidneyswere weighed and comparativeresults were given in fig.1 for non-diabetic groups and fig. 2 for diabetic groups induced with alloxan. Also glucose level in organs were measured for each group and the results were shown in fig. 3 for non-diabetic groups and in fig. 4 for non-diabetic groups and also comparative results of each plant were shown infig. 5-8.
In non-diabetic groups, the results were indicated that there were no significant differences between brain and heart weights when compared with control (P>0.05).Liver weights were lower than control but onlyUD and VA treat-ed groups were significantly differentas compartreat-ed to other plants and control (P<0.05). Moreovercomparison of all non-diabetic groups demonstrated thatVA and EG extracts weresignificantlydecreased kidney weights (P<0.05) (fig. 1).
Fig. 1: Effects of the tested plant extracts on organ weights in
non-diabetic groups
(UD:Urticadioica,VA:Viscum album, EG: Eucalyptus globulus, OE: Olea europea)
Experimental results of diabetic groups showed that there were no significant differences between weights of brain, heart and kidney as compared with synthetic drug, glutril (P>0.05).When compared the liver weights of all tested groups, a significant <0.05 was found between glutril and EG treated groups (fig. 2).
Fig. 2:Effects of the tested plant extracts on organ weights in
diabetic groups
(UD:Urticadioica,VA:Viscum album, EG: Eucalyptus globulus, OE: Olea europea)
Results indicated that no significant changes in glucose levels of brain, heart and kidney were observed between non-diabetic groups and diabetic groups (P>0.05).When compared the glucose level of livers withextract treated and control groups, glucose levels of plant extract treated groups were lower than control but there were no signifi-cant difference between these non diabetic groups. Com-parison of liver glucose levels of plants and glutril treated diabetic groups showed that all tested groups reduced
TABLE 2: The effect of tested plant extracts on blood glucose levels and body weights
Study Groups Blood glucose level (mean±SD) (mg/dl) Body weight (g) Initially Finally Initially Finally Non-diabetic groups Control 83.9±12.1 93.2±2.4 27.9±3.7 25.3±2.9
UD 127.5±3.5 85.0±2.8 20.0±1.4 19.0±2.8 VA 126.0±5.7 82.0±4.2 18.5±2.1 19.0±1.4 EG 107.0±2.8 67.5±3.5 19.0±0.2 18.5±1.2 OE 100.5±6.4 74.5±3.5 19.5±2.1 19.0±2.7 Alloxan induced
diabetic groups GlutrilUD 349.8±13.8374.6±5.8 96.3±2.3*90.9±3.9* 30.4±4.530.7±3.9 26.0±4.227.6±4.2 VA 357.5±9.6 95.2±2.4* 31.0±3.9 26.4±3.6 EG 372.2±12.3 75.3±3.1* 29.9±3.5 25.8±3.3 OE 363.9±23.4 93.4±2.7* 28.7±4.9 24.3±3.3
glucose levels in liver but results not found significantly different. In addition to these results when glucose levels of liver compared with all tested groups, a significant <0.05 was found between diabetic and non-diabetic groups(fig. 3-4).
Fig. 3: Effects of the tested plant extracts on organ glucose levels in
non-diabetic groups
(UD:Urticadioica,VA:Viscum album, EG: Eucalyptus globulus, OE: Olea europea)
Fig. 4: Effects of the tested plant extracts on organ glucose levels
in diabetic groups
(UD:Urticadioica,VA:Viscum album, EG: Eucalyptus globulus, OE: Olea europea)
When compared the each plants extract treated both dia-betic and non-diadia-betic groups, control and glutril between each other for effect of reducing organ glucose levels ma-jor changes determined in liver glucose levels (<0.05)(fig. 5-8). When compared with the control and glutril the results showed that EG and OE were the most effective plants re-ducing organ glucose levels (fig. 7-8).
Fig. 5: Effects of the UD extracts on organ glucose levels in both
tested groups compared with control and glutril (UD:Urticadioica)
Fig. 6:Effects of the VA extracts on organ glucose levels in both
tested groups compared with control and glutril (VA:Viscum album)
Fig. 7:Effects of the EG extracts on organ glucose levels in both
tested groups compared with control and glutril (EG: Eucalyptus globulus)
Fig. 8:Effects of the OE extracts on organ glucose levels in both
tested groups compared with control and glutril (OE: Olea europea)
DISCUSSION
Many studies have been reported on hypoglycemic ef-fect of plants used as anti-diabetics[35]. Literature data in-dicated that all tested plants in this studytraditionally used for treatment of various illness in folk medicineincluding diabetes[16,22,27,32]. Therefore we focused on the bio-chemical role of four medicinal plants UD, VA, EG and OE which possess anti-diabetic potential and aqueous extracts of the plants were tested for their antidiabetic actions and glutril used as positive controlin this study. To determine the comparative effects of tested plants on blood glucose levels, body weights, organ weights (liver, brain, kidney, heart) and organ glucose levels were examined in diabetic and non-diabetic mice.
lev-els in normal values (93.2±2.4-67.5±3.5 mg/dl) in non-diabetic groups. The plants decreased the blood glucose concentrations significantly just as glutril in alloxan induced diabetic groups (P<0.001).(UD:90.93.9; VA:95.22.4; EG:75.3±3.1; OE:93.7±2.7; glutril: 96.32.3 mg/dl). The comparative results of blood glucose levels indicated thatEGwas the most effectiveness plant.When compared the diabetic and non-diabetic groupsthere were no signifi-cant changes observed in body weights(>0.05).Whereas the liver weights ofUD and VA treated groups andkidney weights of VA and EG treated groups were significantly different as compared to other plants and control in non-diabetic groups (P<0.05). There were no significant differ-ences between weights of brain, heart and kidney (P>0.05), while liver weights of glutril and EG treated groups showed significant difference (<0.05) when compared with glutri-land alloxan induced diabetic groups.Glucose levels of liver compared with all tested groups, a significant <0.05 was found between diabetic and non-diabetic groups while no significant changes in glucose levels of other tested organs were observed in both tested groups (P>0.05).
In this study, we were planned to examine the changes inblood and liverglucoseconcentrations according to periods but,suddendecrease inthe amount ofblood glucose levelsled us studytheglucose levels ofbrain, heart, kidneys. Fig. 2 and 3 showed us that thehighestglucoseconcentrationwas found in the liver. Organglucose levels seemed to differwithin the group. Althoughheart is a continuouslyworkingorgan, glu-cose concentrationin the control group was 84.02.2 mg/dl. Itseemsto be a contradiction however, it is an expected val-ue. As heart muscles work both isotonicandisometric, a large part ofthe energyis known toprovide from the beta-oxidation of fatty acidsand creatinephosphate.Glucose is stored in the liver so, change between glucose and glycogen is mostly seen in liver[36]. This may cause high level of glucose in liver. On the other hand, theconcentration ofglucose inkid-ney was as expected. Brain isthe most glucose andfructose usingorgan and glucose concentration ofthe brain depends ontransport of glucose from blood-brainbarrier to blood[36]. That is why, brainglucose concentrationwas lower than liver.
Antidiabetic activitiy of secondary metabolites includ-ing flavonoids, polyphenols, terpenoids, coumarins and al-kaloids were reported by many researcher. Furthermore, in several studies hypoglycemic actions of flavonoids includ-ing hesperidin, quercetin and luteolin, and terpenoids were determined[37]. The all tested plants contained similar com-ponents and especially phenolicswere common constituent of them[14,19,25,29].As major components EGcontained -pinene, -pinene, terpenes and tannins[38] and VA con-tained viscotoxins however, effects of viscotoxins on glu-cose levels were not known. It was probable that viscotoxins showed potential inhibitory effect on glycogenolysis[39-41]. OE contained several constituents such as oleuropeoside in-cluding oleuropein and verbascoside, and flavonoids[29]. UD contained butyric acid, caffeic acid and phenolic acid which was thought to cause acidosis and decrease glucose levels[39,42,43].
Different biological activities were reported for tested plants which related to their secondary metabolites and an-tidiabetic activity[12,21,28-30]. The anan-tidiabetic activity of four tested plants were examined in several diabetic animal models induced by streptozotocin or alloxan in different stud-ies[44-47].Leaf extractprepared from an alcoholic “mother tincture” of E. globuluswas found to significantly reduce the blood glucose levelin alloxaninduceddiabetic rats[44],
anti-diabetic activity of aqueous extract obtained from O. euro-paea[48] and methanol extract obtained fromV. album[49] and hypoglycemic activity of lectin isolated from Urtica species[42]were showed instreptozotocin induced diabetic rats. Literature data mentioned above were indicated that all these results supported our findings inalloxaninduceddiabet-ic minalloxaninduceddiabet-ice. Addition to determination of antidiabetinalloxaninduceddiabet-ic effect of tested plants in diabetic mice,this researchcompared effects of tested plants with each other by using changes on blood glucose levels, body weights and glucose levels and weight of selected organs.
CONCLUSIONS
The results of this study showed that four tested plants reduced blood glucose levels in alloxaninduced diabetic mice and loss of body weight not observed in plant extract treated groups.The aqueous extracts of all tested plants seem todecrease fasting blood glucose levelssimilar to antidia-betic agent glutril. Also no adverse effects were determined in plant extracts treated groups.As all available drugs have undesirable adverse effects, we believe that different medi-cal plants can be used to decrease hyperglycemia. Based on the present findings further investigation is necessary to de-termine constituents which responsible from antihypergly-cemic activity and mechanisms of actions to consider these plants as alternative therapeutics and in the light of future studies, these plants can be considered as alternative thera-peutics.
ACKNOWLEDGEMENTS
Authors would like to add special thanks to SaadetDayıoğluÇelikfrom Dumlupinar University, Faculty of Arts and Sciences, Department of Biology, Kutahyafor their contributions to this study.
REFERENCES
[1] Medagama AB,BandaraR.2014. The use of complementary and alternative medicines (CAMs) in the treatment of diabetes mellitus: is continued use safe and effective?Nutr J, 13(102):1-9.
[2] Ghorbani A, Shafiee-Nick R,
RakhshandehH,BorjiA,2013. Antihyperlipidemic Effect of a polyherbalmixture in Streptozotocin-Induced diabetic rats. Journal of Lipids, 2013:1-6.
[3] ShrivastavaSR, ShrivastavaPS,Ramasamy J,2013. Role of self-care in management of diabetesmellitus. JDia-betes MetabDisord, 12(14):1-5.
[4] Marques AM, Pereira SL, Paiva RA, Cavalcante CV, Sudo SZ, Tinoco LWet al. 2015 Hypoglycemiceffect of themethanolflowerextract of Piperclaussenianumandthema-jorconstituent 2’,6’-dihydroxy-4’-methoxychalcone in streptozotocindiabeticrats. Indian J PharmSci,77(2):237-42.
[5] Elekofehinti OO, 2015. Saponins: Anti-diabetic principles from medicinal plants-A review.Pathophysiol-ogy,22:95-03.
[6] BahmaniM, Zargaran A, Rafieian-KopaeiM,Saki K, 2014. Ethnobotanical study of medicinal plants used in the management of diabetes mellitus in the Urmia, North-west Iran.AsianPac J Trop Med, 7(Suppl 1):348-54.
[7] GhazanfarK, GanaiBA, Akbar S, MubashirK., Dar SA, Dar MYet al.2014. Antidiabetic activity of Artemisia amygdalinaDecne in Streptozotocin induced diabetic rats.
[13] Patel SS,UdayabanuM,2014. Urticadioica extract attenuates depressive like behavior and associative memory dysfunction in dexamethasone induced diabetic mice.Metab Brain Dis,29:121-30.
[14] Orcic D, FranciskovicM, BekvalacK, SvircevE, Beara I, Lesjak Met al.2014. Quantitative determination of plant phenolics in Urticadioicaextracts by high-performance liquid chromatography coupled with tandem mass spectro-metric detection. Food Chem, 143:48-53.
[15] Gülçin I, Küfrevioğlu OI,
OktayM,BüyükokuroğluME,2004. Antioxidant, anti-microbial, antiulcer and analgesic activities of nettle (UrticadioicaL.).J Ethnopharmacol,90:205-15.
[16] Qujeq D, Tatar M, FeiziF, ParsianH, FarajiAS,HalalkhorS.2013. Effect of Urticadioicaleaf alco-holic and aqueous extracts on the number and the diameter of the islets in diabetic rats.Int J Mol Cell Med, 2:21-6.
[17] Yilmaz B, Basar O, Aktas B, Altinbas A, Ekiz F, Büyükcam F et al. 2014. Effects of Urticadioicaextract on experimental acute pancreatitis model in rats.Int J Clin-ExpMed, 7:1313-8.
[18] Binaii M, Ghiasi M, Mohammad S,Farabi V, Pourgholam R, Fazli Het al.2014. Biochemical and hemato-immunological parameters in juvenile beluga (Husohuso) following the diet supplemented with nettle (Urticadioica). Fish Shellfish Immun, 36:46-51.
[19] PietrzakW, Nowak R,OlechM, 2014. Effect of ex-traction method on phenolic content and antioxidant activ-ity of mistletoe extracts from Viscum album subsp. Abietis. Chem Pap, 68:976-82.
[20] Choudhary MI,Maher S,Begum
A,AbbaskhanA,Ali S, Khan A et al. 2010. Characterization and antiglycationactivity of phenolic constituents from Vis-cum album (European Mistletoe).Chem Pharm Bull,58:980-2.
[21] Orhan DD, SezerSenol F, HosbasS,Erdogan Orhan I. 2014. Assessment of cholinesterase and tyrosinase inhibi-tory and antioxidant properties of Viscum album L. samples collected from different host plants and its two principal sub-stances. Ind Crop Prod, 62:341-9.
[22] AmerB ,JuvikOJ, DupontF, Francis GW,FossenT,2012. Novel aminoalkaloids from European mistletoe (Viscum album L.).Phytochem Lett,5:677-81.
[23] OluwaseunAA,GaniyuO, 2008Antioxidant prop-erties of methanolic extracts of mistletoes (Viscum album) from cocoa and cashew trees in Nigeria.Afr. J. Biotechnol, 7:3138-42.
[24] Akolade JO, Olajide OO, AfolayanMO, Akande SA, Idowu DI,Orishadipe AT,2012. Chemical composition, antioxidant and cytotoxic effects of Eucalyptus globulus-grown in north-central Nigeria.J. Nat. Prod. PlantResour, 2:1-8.
[25] Topiar M,Sajfrtova M, Pavela R, MachalovaZ, 2015. Comparison of fractionation techniques of CO2 ex-tracts from Eucalyptus globulus-composition and insecti-cidal activity.J. of Supercritical Fluids, 97:202-10.
[26] Santos GG, Alves JCN, RodillaJML, Duarte AP, Lithgow AM, Urones JG,1997. Terpenoids and other con-stituents of Eucalyptus globulus.Phytochemistry, 44:1309-12.
[27] Song A, Wang Y,LiuY,2009. Study on the chemi-cal constituents of the essential oil of the leaves of Euchemi-calyp- Eucalyp-tus globulusLabill from China.Asian J Tradit Med, 4:134-40.
[28] KanojiyaD, ShankerD, Sudan V, Jaiswal
AK,ParasharR.In vitro and in vivo efficacy of extracts of leaves of Eucalyptus globulus on ovine gastrointestinal nematodes. Parasitol Res. 2015;114:141-8.
[29] Laham SAA,Fadel FMA,2014. Antibacterial ac-tivity of various plants extracts against antibiotic-resistant Aeromonashydrophila. Jundishapur J Microbiol, 7:1-7.
[30] Yuan J, Wang C, Ye J, Tao R,Zhang Y,2015. En-zymatic hydrolysis of oleuropein from Olea europea(Olive) leaf extract and antioxidant activities.Molecules, 20:2903-21.
[31] Lee-Huang S, Zhang L, Huang PL, Chang Y, Huang PL.,2003. Anti-HIV activity of olive leaf extract (OLE) and modulation of host cell gene expression by HIV-1infection and OLE treatment.BiochemBiophys Res Com-mun, 307:1029-37.
[32] Cumaoglu A, Ari N, Kartal M,KarasuC,2011. Polyphenolic extracts from Olea europea L.protect against cytokine-induced beta-cell damage through maintenance of redox homeostasis. Rejuvenation Res, 14:325-34.
[33] Botsoglou E, Govaris A, AmbrosiadisI, FletourisD,BotsoglouN,2014. Effect of olive leaf (Olea europea L.) extracts on protein and lipid oxidation of long-term frozen n-3 fatty acids-enriched pork patties. Meat Sci, 98:150-7.
[34] ColowickSP, Kaplan ON.,1965. Methods in Enzy-mology. New York: Acad. Press Inc. Publishers;.
[35] Patel DK, Kumar R, LalooD,HemalathaS,2012. Natural medicines from plant source used for therapy of dia-betes mellitus: An overview of its pharmacological aspects. Asian Pac J Trop Dis, 2:239-50.
[36] Nelson DL,Cox MM. 2004. Lehninger Principles of Biochemistry, 4th. ed. New York:W.H. Freeman and Company.
[37] Afifi-YazarFU, Kasabri V,Abu-Dahab R,2011. Medicinal plants from Jordan in the treatment of diabetes: traditional uses vs. in vitro and in vivo evaluations-part 2. Planta Med, 77:1210-20.
[38] OkpashiVE,Bayim BPR,Obi-AbangM, 2014. Comparativeeffects of somemedicinalplants: Anacardiu-moccidentale, Eucalyptusglobulus, PsidiumguajavaandXy-lopiaaethiopicaextracts in alloxan-induceddiabeticmalewis-tar albino rats.BiochemResInt, 2014:1-13.
[39] Moore M, 1995.Plant Materia Medica 5.0. South-west School of Botanical Medicine.
[40] CorderoCM,LázaroM, AgudoMA,Navarro E, Tru-jillo J,AyusoMJ, 2001.A cytotoxic diarylheptanoid from Vis-cumcruciatum. Phytochemistry, 58:567-9.
[41] MillsS,1994. The Complete Guide to Modern Plantism. New York: Thorsons Press.
[42] Kavalalı G, Tuncel H, Goksel S, Hatemi HH, 2003. Hypoglycemic activity of Urticapilulifera in strepto-zotocin-diabetic rats. J Ethnopharmacol, 84:241-5.
[43] Hoffmann D, 1990. The New Holistic Plant, 2nd ed. Shaftesbury:Element.
[44] AhlemS, Khaled H, WafaM, SofianeB, Mohamed D, Jean-Claude M et al.2009. Oral administration of Euca-lyptus globulusextract reduces the alloxan-inducedoxidative stress in rats.ChemBiol Interact, 181:71-6.
[45] ShahaboddinME, PouramirM, MoghadamniaAA, LakzaeiM, Mirhashemi SM,MotallebiM,2011. Antihyper-glycemic and antioxidant activity of Viscum album extract. Afr J PharmPharmacol, 5:432-6.
[46] OzkolH, TuluceY, DilsizN,KoyuncuI, 2013. Ther-apeutic potential of some plant extracts used in Turkish tra-ditional medicine on Streptozocin-Induced Type 1 Diabetes
Mellitus in rats. J MembraneBiol, 246:47-55.
[47] Said O, FulderS, Khalil K, Azaizeh H, KassisE,SaadB, 2008. Maintaining a physiological blood glucose level with ‘glucolevel’, a combination of four anti-diabetes plants used in the traditional Arab herbal medicine. JEvidBased Complementary Altern Med, 5;421-8.
[48] Amin ME, Virk PM,ElobeidAR, Almarhoon ZM, Hassan ZK, Omer SAet al. 2013. Anti-diabetic effect of Murrayakoenigii(L) and Olea europaea(L) leaf extracts on streptozotocin induced diabetic rats.Pak J PharmSci, 26:359-65.
[49] AdaramoyeO, AmanlouM, Habibi-RezaeiM, Pas-alar P, Moosavi-MovahediA, 2012. Methanolic extract of African mistletoe (Viscum album) improves carbohydrate metabolism and hyperlipidemia in streptozotocin-induced diabetic rats. Asian Pac JTrop Med, 5:427-33.
