34 Planta Med.
57(1991)
Antiinflammatory, Analgesic, and Antipyretic Effects of
an Aqueous Extract of Erythraea centaurium
Tayfunflerkan1'2. Levent Ustilnes', Ferzan Lermioglu', andAsli Ozer'
1
Department of Pharmacology, Faculty of Pharmacy, Ege University, Izmir, Turkey
2 Address for correspondence
Received: November 10, 1989
Abstract
Erythraea centauriu,n is a plant which is
used in the treatment of various inflammatory conditions
in popular medicine. The aqueous extract of the plant
has been examined for its antiinflammatory, analgesic,
and antipyretic effects in several animal models. The ex-
tract exhibited antiinflammatory and antipyretic activity
although no analgesic activity was observed.
Key words
Erythraea centauriurn, Gentianaceae, an-
tiinflammatory effects, analgesic effects, antipyretic ef-
fects.
Introduction
Erythraea centauriurn (Gentianaceae) is
known in Turkey as "Red Cantarone". The plant has a wide-
spread use in popular medicine against different types of in-
flamed wounds as a wound healer. Usually it is collected in
bloom during the flowering season. Its herbal part is boiled
with water and a small cup is drunk daily for treating the
chronic inflamed wounds. The salve of the plant which is
prepared with olive oil, is used externally for the same pur-
pose.
In spite of an extensive search, it was not
possible to find any literature concerning the related phar-
macological actions of the plant. The present study was,
therefore, undertaken to work out whether the aqueous ex-
tract of the plant exerts antiinflammatory, analgesic, and
antipyretic actions.
Materials and Methods
Preparation of the extract
Erythraea centaurium is a herbaceous plant
reaching 20—50cm in height which usually grows at forest edges
and humid meadows. Our samples were collected from Karabui-un —
Izmir and were identified at the Pharmaceutical Biology Depart-
ment of the Faculty of Pharmacy of Ege University. After identifica-
tion the samples were dried in the shade at ambient temperature
(25—30°C). The dried plants were roughly cut and ground to pow-
der. The weighed powder (230 g) was macerated with distilled
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water until the macerate lost its colour. The resulting suspension
was squeezed through several layers of cotton gauze to remove
coarse particles and evaporated in a rotating evaporator under re-
duced pressure until dryness. Each g of the dry extract obtained by
this procedure was equivalent to 3.83 g of the plant powder and
was dissolved in 0.9 % NaCI solution at appropiate concentrations
for pharmacological tests.
Antiinflamrnatory activity
In view of the importance of this activity, the an-
tiinflammatory property of the extract was investigated in two ani-
mal models of subchronic inflammation (1) in comparison with a
number of well established reterence compounds.
One was the air pouch granuloma originally de-
scribed by Selye (2) and modified by DiPasquale et al. (3). Male,
Swiss Albino rats (200—300g) were individually housed and ar-
ranged in groups as indicated in Table 1. The rats were lightly
anesthetized with ether and 20 ml of air were injected s.c. into the
shaved dorsal surface of the animals forming a pocket. 0.5 ml of 1 %
croton oil in sesame oil was then injected into the air pouch. Control
cream and the extract at different concentrations in creams were
applied topically to the surface of the pouch and spread over a con-
trolled area (35mm in diameter) daily for 8 days starting on day 2.
On day 4 all the pouches were reshaved and slightly reinflated to
original turgidity and 0.5 ml of a 3% croton oil solution was injected
directly into the pouchs. All animals were killed on day 10 and the
exudate volume was measured. The final body weights and the ad-
renal and thymus weights were also recorded.
The second model used was the polyarthritis, in-
duced by the intradermal injection of Freund's adjuvant (4). The
adjuvant was purchased from Sigma (1 mg/mi Mycobacterium
tuberculosis). Swiss Albino rats of either sex weighing approxi-
mately 200—250 g were used and were housed three in a cage. On
day 0, the animals were given a single 0.1 ml intradermal injection
of the adjuvant into the subplanter area of the right hind paw. In-
domethacin and diklofenac sodium which were used as positive
controls were administered by oral route on a mg/kg body weight
basis. The extract dissolved in 0.9% NaC1 solution was given di-
rectly by the same route at the doses of 1, 5, 10, 50, 100, and
500mg on days 12 and 16 following adjuvant injection. Thickness
of both hind-feet were measured with a micrometer on day 0 before
adjuvant injection, on day 12 and approximately three hours after
final drug administration on day 16. The body weights of the rats
was also recorded on the same days.
Antiinflainmatory. Analgesic, and Antipyretic Effects of an Aqueous Extract ofErythraea centauriurn Planta Med. 57(1991) 35

Analgesic activity
The analgesic activity of the extract was assessed
in two well established models, the writhing syndrome of the
mouse and the hotplate test.
In the first model the writhing syndrome was eli-
cited by i.p. injection of 3% acetic acid at the dose level of 300 mgI
kg (10 mi/kg), 30 and 60 mm after 1, 5, 10, 50, and 100mg ip. in-
jection of the extract. The number of writhes were noted for the
10 mm period between 5 and 15 mm after acetic acid injections (5).
In the other model, the analgesic activity was
tested in mice (20—30 g) with the hotplate method (53.5°C), 30 and
60mm after 1, 5, 10, 50, and 100mg i.p. injection of the extract.
The time when the aminals licked either of their hindpaws was ac-
cepted as end-point. The cut- off time was 60 sec (6).
Antipyretic activity
The antipyretic effect of the extract was tested in
basic body temperatures. The temperatures of the same animals
were recorded 0.5, 1, 2, 3, and 4h after DNP administration. The
maximum hyperthermia of DNP was observed 0.5 h after its injec-
tion, therefore, the extract was administered to the animals in dif-
ferent concentrations i.p. 0.5 h after DNP injection. The rectal body
temperatures of the animals were recorded at the same time inter-
vals mentioned above.
In the third model, d-amphetamine sulphate was
administered i.p. to rats fasted for 12 h in a dosage of 10mg/kg (9).
The continuation of the test procedure and the temperature mea-
surement time intervals were as in DNP hyperthermia.
Results and Discussion
Antiinflammatory effects
The results of the air-pouch granuloma
bioassay procedure are given in Table 1. In spite of the ad-
vantages and the disadvantages of the method for evaluat-
ing topically applied antiinflammatory preparations as dis-

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a standard screening for compounds exhibiting this activity. Male,
Swiss Albino rats weighing 200—2 50 g were utilized in all experi-
ments and the rectal body temperatures were measured using an
applicator type Ri by means of an Electrical Universal Thermome-
ter of the TE3 Ellab type.
In the first test carried out, the body tempera-
tures of the rats which were fasted overnight with water ad lib.
were taken initially at time zero. The animals were weighed and in-
jected s.c. in the neck region with either of the following: a 15% sol-
ution of brewer's yeast or 0.9% NaC1 solution (10 mi/kg) (7). Tem-
peratures were again taken 2.5 h after the injection of yeast. The in-
itial readings and the 2.5 h readings were combined to form the in-
itial temperature observation. 4 h after the yeast injection, the rats
were given extract suspensions by gavage. Temperature readings
were repeated at 5 and 6 h after the yeast injection.
2,4-Dinitrophenol (DNP) prepared at a concen-
tration of 1 mg/ml in 0.9% NaCl solution was administered i.p. as a
pyretic agent in a dosage of 20mg/kg in the second model in order
to obtain experimental hyperthermia (8). DNP was administered to
the animals fasted overnight with water ad lib. after checking the
cussed by DiPasquale et al. (3) and Fisher (10) our findings
clearly indicate that 10% extract cream at the dose of
200 mg/day exhibits a significant decrease in exudate for-
mation. 2.5% and 5% extract creams inhibited the exudate
formation by aproximately 19% and 42%, respectively.
The results of the adjuvant induced polyar-
thritis which was tested therapeutically by administrating
the extract at different doses when the edema had begun to
develop are presented in Table 2. Although less than the
known drugs indomethacin and dikiofenac, it is quite evi-
dent that the extract was effective in reducing the edemat-
ous phase of the polyarthritis at doses of 10 to 500 mg/day
in this system. The anti-edematous doses of the extract
were also effective in counteracting the body weight loss
which was normally observed in the rats with adjuvant dis-
ease.

Table 1 Effects of the extract and drugs on the

Daily Average % Rel. Organ Weights A B. W}
air-pouch granuloma.
Treatment N Cream Exudate Inhib. mg/loOgbodyweight (g)
Dose ml S.D.
mgi Adrenal Thymus
animal

Control untreated 7 — 18.0 0.91 0 0.019 0.166 + 35

Creamcontrol 7 200 17.9 0.57 0.5 0.020 0.170 +28
Etofenamate(5OmgIg)gel 6 100 7.9 0.73" 56 0.020 0.160 + 18
Clobetasol 17-butyrate 6 50 8.6 0.46" 52 0.016 0.075 +6
(0.05%) cream

Diethylaminesalicylate 6 200 4.8 0.39" 73 0.020 0.105 + 12

(4 g/40 g( cream

Extractcream(l%) 7 200 15.1 0.51' 16 0.019 0.139 +24

Extractcream(2.5%( 6 200 14.7 0.58" 19 0.021 0.143 + 19
Extract cream (5 %) 7 200 10.5 0.61" 42 0.020 0.113 + 19

Extractcream(10%( 6 200 9.5 0.33" 47 0.022 0.110 + 16

Extract (pure) 6 200 17.4 0.92 3 0.021 0.156 + 28

a No. of rats.
Change in body weight from initial weight.
** Difference (P < 0.01) from controls;' = difference (P < 0.05( from controls.
36 Planta Med. 57(1991) Tayfun Berkan eta!.

Table 2 Effects of the extract and two known an-

Average Thickness
tiinflammatory drugs on rats with adjuvant disease.
Treatment Dose of the Injected % Inhibition Average Body
Foot of Thickening Weight Gains
mm±S.D. g±S.D.
Control — 6.138 0.20 — 5.8 2.0
Indomethacin 1 mg/kg/day 1.810 0.34" 70.5 28.0 5.7"
Dikiofenac-Na 4mg/kg/day 2.608 0.22" 57.5 17.0 2.7"
Extract 1 mg/day 5.988 0.03 2.4 6.7 2.5
Extract 5mg/day 5.900 006' 3.9 6.7 2.5
Extract 10mg/day 5.632 0.04" 8.3 12.5 5.2'
Extract 50mg/day 5.189 0.14" 15.5 15.0 3.1"
Extract 100 mg/day 4.871 0.21" 20.6 16.6 4.0"
Extract 500 mg/day 4.302 0.16" 29.9 19.1 3.7"

Difference (p <0.05) from adjuvant control;• Difference (p <0.01) from adjuvant control, (n = 6).

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Table 3 Effects of the extract on yeast-induced
°C±S.E.
Treatment Dose hyperthermia.
(/.p.) Initial 5h 6h

Yeast Control — 36.7 0.11 37.9 0.10 38.1 0.08

Indomethacin 5mg/kg 37.1 0.03' 37.4 0.07'
Extract 1mg 37.7 0.08 37.9 0.02
Extract 5mg 37.7 0.07 37.9 0.08
Extract 10mg 37.6 0.07 37.9 0.05
Extract 50mg 37.4 0.10' 37.7 0.17
Extract 100mg 37.3 0.13' 37.6 0.11'
Saline Control — 36.8 0.05 36.9 0.03 36.8 0.03
Extract 100mg 36.9 0.06 36.9 0.03
Difference from yeast control (p <0.05), (n = 7).

Table 4 Effects of thee xtract on 'DNP' induced hyperthermia.

°C± SE.
Treatment Dose
Initial 0.5h 1.Oh 2.Oh 3.Oh 4.Oh

2,4-Dinitrophenol
(control) 20mg/kg 36.8 0.19 39.3 0.12 39.0 0.13 38.5 0.15 38.1 0.14 37.5 0.21
Extract 1mg 37.0 0.07 — 39.8 0.06 38.7 0.05 38.5 0.04 38.3 0.05
Extract 5mg 37.0 0.02 — 39.1 0.05 38.5 0.05 38.3 0.06 38.1 0.04
Extract 10mg 36.9 0.04 — 39.3 0.06 38.3 0.08 38.2 0.07 38.1 0.05
Extract 50mg 36.9 0.06 — 38.4 0.04* 38.1 0.05* 37.5 0.02' 37.1 0.03
Extract 100mg 36.9 0.06 — 38.1 0.10* 37.6 0.07' 37.2 0.02* 37.0 0.01'
* Difference from DNP control (p <0.05), )n = 9).

Analgesic effects Antipyretic effects

The extract did not show any analgesic ac-
tivity in mice in both methods tested in any of the doses ad-
ministered i.p.. This result may be dissapointing because
antiinflammatory compounds usually have analgesic and
antipyretic properties also. Therefore, this activity should
be tested again by changing the route of administration,
dosage, and the time intervals.
The extract reduced yeast-induced hyper-
thermia at the dose of 100 mg. This dose did not produce a
hypothermic effect in saline injected rats (Table 3).
When administered at the maximum of
DNP effect, the extract decreased the increased tempera-
ture of the rats at the doses of 50 and 100mg. This effect
Antiinflammatorg, Analgesic, and Antipyretic Effects of an Aqueous Extract ofErythraea centaurium Planta Med. 57(1991) 37

Table 5 Effects of the extr act on amphetamine -induced hyperthermia.

°C±S.E.
Treatment Dose
Initial 0.5h 1.Oh 2.Oh 3.Oh 4.Oh

Amphetamine (control) 10 mg/kg 36.8 0.15 38.9 0.20 39.7 0.20 38.9 0.16 37.7 0.09 37.5 0.07
Extract 1.0mg 37.0 0.07 — 39.3 0.09 39.0 0.05 38.2 0.08 37.6 0.07
Extract 5.0mg 36.9 0.03 — 39.1 0.09 38.3 0.08' 38.0 0.05 37.8 0.08
Extract 10 mg 36.5 0.08 — 39.8 0.12 38.5 0.34 38.0 0.05 37.5 0.07
Extract 50 mg 37.0 0.05 — 39.2 0.06' 38.3 0.06' 37.7 0.09 37.0 0.01'
Extract 100 mg 36.8 0.02 — 38.8 0.04' 38.0 0.02 37.2 0.05' 37.0 0.02'
Difference (p <0.05) from amphetamine control, (n = 9).

Table 6 Effects of the extract pretreatment on

°C±S.D.
DNP and amphetamine hyperpyrexia.'
Treatment
fiSh aoh
______ Initial LOh 3.Oh 4.Oh

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I

2,4-Dinitrophenol 36.9 0.40 39.9 0.18 39.3 0.19 39.1 0.16 39.0 0.41 38.7 0.17
Amphetamine 36.5 0.27 39.3 0.26 39.5 0.31 38.9 0.48 38.0 0.17 36.9 0.18
The extract (100 mg) was given i.p. 30 mm before DNP and amphetamine, (n = 9).

was lasting and was observed in hourly repeated measure- References

ments up to the fourth hour (Table 4). The results obtained
from the experiments carried out with amphetamine were
identical to those of DNP (Table 5). Extract pretreatment did
not prevent the temperature increases after DNP and am-
phetamine administrations (Table 6).
The findings of the experiments concerning
the effect of the extract on body temperature indicate that
the extract does not prevent the development of hyperther-
mia and has no effect on normal body temperature but in-
fluences the fever which is already manifested.
1 Berkan, D., Evinc, A. (1972) Aegean Med. J. 1.47.
2
Selye, H. (1953) Proc. Soc. Exptl. Biol. Med. 82, 328.
DiPasquale, G., Rassaert, C. L., McDougall. E. (1970) J. Pharm. Sd.
59, 267.
Newbould, B. B. (1963) Brit. J. Pharmacol. 21, 127.
Chapman, D. B., Way, E. L. (1982) Brit. J. Pharmacol. 75, 389.
6
Benedek, G., Szikszay, M. (1984) Pharrn. Res. Commun. 16, 1009.
Gubler, H. U., Baggiolini, M. (1978) Scand. J. Rheum. 7(21), 8.
8
Kulcsar-Gergely, J. (1976) Arz. Forsch. 26, 55.
Mantegazza, P., Naimzade. K. M., Riva, M. (1968) Eur. J. Pharmacol.
° 4,25.
Fisher, W. J. (1961) J. Pharm. Exp. Ther. 131, 1.

In conclusion, the results of this study

shows that the plant extract exhibits antiinflammatory and
antipyretic activity, therefore, Erythraea centaurium
should be further studied either in its chemical or phar-
macological aspects.