Effects of two Turkish medicinal plants Artemisia herba-alba a n d ...

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Effects of two Turkish medicinal plants Artemisia herba-alba a n d ...

Journal of Cell and Molecular Biology 5: 19-24, 2006.

Haliç University, Printed in Turkey.

Effects of two Turkish medicinal plants Artemisia herba-alba a n d

Teucrium polium on blood glucose levels and other b i o c h e m i c a l

parameters in rabbits

Mehmet Iriadam, 1 Davut Musa, 2 Hatice Gümüflhan 3 and Füsun Baba 4

1 University of Harran, Faculty of Veterinary Medicine, Department of Physiology, fianl›urfa, Turkey,

2 University of Harran, Faculty of Sciences and Arts, Department of Biology, fianl›urfa, Turkey,

3 University of Istanbul, Faculty of Sciences, Department of Biology, Istanbul, Turkey, 4 University of

Harran, Faculty of Medicine, Deparment of Pathology fianl›urfa, Turkey.

Received 26 May 2005; Accepted 16 December 2005

Abstract

Artemisia herba-alba (Asteraceae) and Teucrium polium (Lamiaceae) are traditional plants that are used for the

conventional therapy of several diseases such as diabetes mellitus and gastrointestinal disorders in Turkey. Our aim

in the present study is to investigate the effects of these two plants on blood glucose levels and some other

biochemical parameters to demonstrate their possible therapeutic effects on diabetes. The extracts of aerial parts of

Artemisia herba-alba (85 mg/kg) and Teucrium polium (82 mg/kg) were administered orally to control and

Streptozotocin (STZ)-induced diabetic rabbits. Applied doses did not cause any acute toxicity or behavioural

changes. Blood glucose levels were estimated before and 2, 4, 6 and 8 hours after administration of the extracts. The

Artemisia herba-alba produced a significant (p0.05) effect.

Keywords: Artemisia herba-alba, Teucrium polium, hypoglycaemia, streptozotocin, diabetes.

Türkiye’de yetiflen iki t›bbi bitki Artemisia herba-alba ve Teucrium polium’ u n

tavflanlarda kan glukoz düzeyi ve di¤er biyokimyasal parametreler üzerine etkileri

Özet

Artemisia herba-alba ve Teucrium polium, sindirim sistemi bozukluklar› ve fieker (Diabet) hastal›¤› gibi bir çok

hastal›¤›n tedavisinde geleneksel olarak kullan›lan bölgesel bitkilerdir. Bu çal›flma, bu iki bitkinin fleker hastal›¤›nda

muhtemel tedavi edici etkisini göstermek ve kan glukoz seviyesi ile baz› biyokimyasal parametreler üzerine etkilerini

belirlemek amac›yla yap›ld›. Artemisia herba-alba (85 mg/ kg) ve Teucrium polium’dan (82 mg/kg) elde edilen

ekstraktlar, Streptozotocin (STZ) ile diyabet oluflturulan grup ve kontrol grubu tavflanlara a¤›z yoluyla verildi.

Uygulanan dozlar herhangi bir akut toksisiteye veya davran›fl de¤iflikliklerine yol açmam›flt›r. Kan glukoz seviyeleri,

ekstraktlar›n uygulanmas›ndan önce ve uygulamadan 2, 4, 6 ve 8 saat sonra ölçüldü. Teucrium polium’un glukoz

seviyesi üzerine etkisi önemsiz iken, Artemisia herba-alba kontrol ve diabetik tavflanlarda önemli derecede

hipoglisemi oluflturdu.

Anahtar Sözcükler: Artemisia herba-alba, Teucrium polium, hipoglisemi, streptozotosin, diyabet

19


20 Mehmet ‹riadam et al.

Introduction

Since ancient time phytotherapy has been used as folk

medicine to treat various diseases including diabetes

mellitus. More than 400 traditional plants treatments

for diabetes mellitus have been recorded, but only

small numbers of these have received scientific and

medical evaluation to assess their efficacy (Bailey and

Day, 1989, Satyavati et al., 1987).

Diabetes mellitus is characterised by elevated

plasma glucose concentration resulting from insulin

insufficiency and insulin resistance, or both (ADA,

1989).

A r t e m i s i a species (Fam: Asteraceae), (Mossa

1985; Al-Shamaony et al., 1994; Subramoniam et al.,

1996) and Te u c r i u m species (Fam: Lamiaceae)

(Suleiman et al., 1988; Gharaibeh et al., 1988; Tanira

et al.,1996; Konuklugil et al., 1997) are reported to

possess antidiabetic effects and have been used in

many countries of Middle East and Turkey as a herbal

medicine for treatment of diabetes, high blood

pressure and gastrointestinal ailments.

French Ministry of Health and Humanitarian

Action has suspended marketing authorization for

medical products containing extracts from germander

(Teucrium chamaedrys Fam: Lamiaceae). Several

cases of hepatitis have been reported following

administration of germander (WHO 1992, Larrey et

al., 1992). These findings were marked by jaundice

and increased aminotransferase levels in blood serum.

A focal loss in purkinje cells in the cerebellum in rats

treated with T. stocksianum was reported (Tanira et al.,

1996).

These symptoms appeared 3-18 weeks following

after administration. However, there is no detailed

study demonstrating therapeutic effects, acute and

chronic toxicity of these plants.

In our present work, by using experimental

diabetic animal model we investigated the effects of

these plants on diabetes and the possibility of using

them as alternative therapeutic drugs.

Materials and methods

Animal

Adult New Zealand rabbits of both sex weighing

1.350-2.050 kg and 5-6 months old were used. They

were housed in-groups in large cages and fed on green

vegetables and chaw pellets, and allowed tap water ad

libitum. The animals were kept in laboratory for two

months, before starting the experiments for

acclimation.

Plant material and the extract preparation

Aerial part of A. herba alba and T. polium were

obtained from a herbal and folk medicine market in

Sanliurfa Tu r k e y. The plants were botanically

authenticated and the samples were stored in

Herbarium of Science and Art Faculty, Harran

University, Sanliurfa/Turkey.

The aerial parts of A. herba-alba and T. polium

were coarsely powdered and macerated with distilled

water for 16 hours with occasional stirring. The 8.5

mg/ml A. herba-alba and T. polium 8.2 mg/ml have

been prepared from the filtered extract.

Experimental induction of diabetes in rabbits

Diabetes was induced by 65 mg/kg of streptozotocin

(STZ) Sigma ® administered intraperitoneal (i.p.) in

physiologic saline with pH adjusted to 4.5 using citric

acid as described (Palanichamy et al., 1988). After 8

days of STZ administration and 18 h. of fasting, blood

glucose was measured by collecting 3-5 ml of blood

from the marginal ear vein. The glucose levels that

were above 250 mg/ml were included in the

experiments.

Grouping of animals

Diabetic rabbits were divided into 3 groups of 10

animals each. Normal rabbits were divided randomly

into 3 groups of 10 rabbits each and treated as follows:

A- Groups I-III including normal rabbits were

treated as follows.

Group I- received 10 ml/kg tap water and

served as control.

Group II- received 85 mg/kg Artemisia herbaalba

aqueous extract.

Group III- received 82 mg/kg Teucrium polium

aqueous extract.

B- Group IV-VI including diabetic rabbits were

treated as follows.

Group IV- received 10 ml/kg tap water and

served as control

Group V- received 85 mg/kg Artemisia herbaalba

aqueous extract.

Group VI- received 82 mg/kg Teucrium polium

aqueous extract.


Drug administration

The amounts of Artemisia herba-alba and Teucrium

polium extract were calculated for each rabbit on body

weight basis 85 mg/kg and 82 mg/kg respectively, this

was equivalent to 2g/kg of dried plant (Tanira et

al.,1996) and prepared by dissolved in 10 ml of water

(Gharaibeh et al.,1988; Tanira et al., 1996). The water

and extracts were administered to each rabbit using a

stomach tube attached to 20-ml standard syringe. The

tube was inserted into stomach through the

oesophagus, and the plunger of syringe was pressed

slowly and steadily. Drugs were administered at doses

85 and 82 mg/kg body weight to evaluate the chronic

toxicity.

Collection of blood and biochemical determination

Blood was collected from the marginal vein of the

rabbits using 22 gauge sterile syringe from the

marginal vein of the ear, prior and 2, 4, 6, 8 hours after

drug administration. Plasma was separated by

centrifugation at 3000 rpm for 10 minutes. Plasma

glucose levels were then measured by commercial kits

(Boehringer Manheim, Germany) by using a glucose

oxidize method with a HITACHI 917 ® autoanalyser.

Blood plasma was separated by centrifugation at

3000 rpm for 3 minutes. Blood urea, alanin

transaminase (ALT), aspartate transaminase (AST),

creatinine, cholesterol, and trigliserides were

measured using colorimetric and ultraviolet methods

as recommended by the German Society for Clinical

C h e m i s t r y. Reagents were obtained from Sigma-

Aldrich.

Pathological estimation

All animals were sacrificed by dislocation at the end

of the experimental period. Liver, kidney, brain and

medulla spinalis tissues from control and diabetic

treated groups were fixed in Bouin fixative. After

routine tissue processing they were embedded in

paraffin and sectioned 5 μm thick from each block and

stained with haematoxylin-eosin for ordinary light

microscopic studies.

Statistical analysis

The data were subjected to analysis of variance

(Duzgunes et al., 1987) using SPSS 10.0 for Windows

program. The data were analysed with ANOVA in

SPSS. Duncan’s test was used in all data where

appropriate.

Results

There was no mortality in any experimental group

throughout the investigation period. The effect of

aqueous extract of aerial parts of A. herba-alba and T.

polium on blood sugar values on hours 2, 4, 6, and 8

of euglycemic and STZ-induced diabetic rabbits are

shown in Figure 1 and 2 respectively. 82 mg/kg body

weight of T .polium aqueous extract was insignificant

(p>0.5) neither euglycemic nor STZ-induced diabetic

rabbits when compared with control group.

However 85 mg/kg body weight of A. herba-alba

aqueous extract attenuated the hyperglycaemic effect

of STZ-induced diabetic rabbits. The blood sugar

values of STZ-induced rabbit group on 2, 4, 6 and 8

hours were 269.7±4.8, 246.5±5.2, 253.9±9.5 and

271.5±4.4 respectively and the hypoglycaemic effect

of A. herba-alba extracts was estimated to be 36 % of

initial value.

As shown in Table 1 administration of A. herbaalba

aqueous extract resulted in large reductions (p<

0.05) in alanine aminotransferase, aspartate

aminotransferase and urea levels of STZ-induced

diabetic rabbits group and changes in any biochemical

parameters except increasing in urea levels.

Histopathological results

No significant histopathological changes were noted

in the studied organs (liver, heart, kidney, brain and

medulla spinalis. In A. herba alba group after chronic

and acute treatment. However animals treated with

T. polium showed normal histology of liver, heart and

brain, and simple apoptosis in the kidney sections of

treated groups

Discussion

Effects of two Turkish medical plants 21

The aim of this paper is to study the hypoglycemic

effect and the toxicity of A. herba-alba and T. polium

and to approve its traditional usage for controlling

diabetes mellitus or other diseases.

This present study reveals that oral administration

of A. herba-alba aqueous extract showed significant

(p


22 Mehmet ‹riadam et al.

Table 1. The effects of A. herba alba and T. polium on biochemical and haematological parameters (n=10 for 6 groups).

Biochemical STZ untreated STZ untreated STZ untreated STZ treated STZ treated STZ treated

parameters Rabbits received Rabbits Rabbits Rabbits Rabbits Rabbits

10ml/kg tap received received received received received

water (Control) 85mg/kg 82mg/kg 10ml/kg tap 85mg/kg 82mg/kg

A. herba alba T. polium. water (Control) A. herba alba T.polium.

Alanine

Aminotransferase

(U/L-1) 48.7±3.6 49.80±1.9 69.1± 2.3 82.5±6.4 51.2±5.6* 86.0±3.4

Aspartate

Aminotransferase

(U/L-1) 79.9±1.7 78.0±2.6 112.2±3.6 148.4±7.9 89.9±7.9* 147.1±3.1

Cholesterol

(mol/L-1) 4.1±0.3 4.3±0.1 3.9±0.3 6.6±0.6 4.7±0.7 4.2±0.2

Creatinine

(mg/dL-1) 0.6±0.1 0.62±0.1 0.7±0.08 2.21±0.02 0.66±0.09 2.57±0.7**

Tryglycerides

(mg/L-1) 55.3±0.5 53.0±1.4 53.1±0.9 47.9±4.4 56.4±3.2 54.7±2.9

Urea

(mg/dL-1) 21.2±0.7 23.1±2.3 106±2.3 68.9±2.5 26.2±2.4 111±5.2*

Glutamyl

transferase

(Unit/ L-1) 48.7±0.6 50.2±1.8 50.3±2.4 52.1±3.5 51.1±1.1 50.6±2.2

Total protein

(mg/dL-1) 6.71±0.2 7.1±0.5 6.5±0.9 7.2±0.3 6.92±0.2 7.5±1.1

Values are means ± s.d.(n=10)

* p0.05 insignificantly different from control group

Figure 1. Effects of A. herba-alba and T. polium on blood

glucose level in nondiabetic rabbits

treated rabbits, while the effect of T. polium aqueous

extract was insignificant (p>0.05). These results are

similar with those of Twaij and A l - B a d r, 1988),

Al- Shamaony et al., (1994) and Marrif et al.,(1995)

who reported that A. herba alba p r o d u c e d

hypoglycemia in both normal and hypoglycemic

Figure 2. Effects of A. herba alba and T. polium on glucose

levels in STZ-induced hyperglycaemic rabbits

rabbits. Gharaibeh et al., (1988) reported that T.

polium extract caused significant reduction in blood

glucose concentration 4 hours after intravenous (i.v.)

administration and 24 hours after i.p. administration.

Ansari et al., (2000) reported that T. polium did not

show any effect on blood sugar levels in patients.


Our results are at variance with those of Gharaibeh

et al. (1988), but similar with Ansari et al. (2000). It is

well known that sulphonylurea drugs like tolbutimide

lower blood glucose level by stimulating pancreatic βcells

to release insulin in to the blood stream, which

have been reported to produce hypoglycaemia, thus

increasing glycogen deposition in the liver causing

reduction of glucose levels which possibly increase

the insulin receptors. STZ-induced diabetes by

destroying β-cells and impairing renal function.

However in this study tolbutamide exhibited only anti

h y p e rglycaemic animals induced by STZ-diabetic

rabbits (Wadood et al., 1992). However in this study

aqueous extract of A.herba alba (85 mg/kg) showed

significant (p


24 Mehmet ‹riadam et al.

Acknowledgements

This study was partially supported by the Research

Foundation of Harran University.

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