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 Address for correspondence
2
Received: November 10, 1989
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 extract exhibited antiinflammatory and antipyretic activity although no analgesic activity was observed.
Key words Erythraea centauriurn, Gentianaceae, antiinflammatory effects, analgesic effects, antipyretic effects.
der. The weighed powder (230 g) was macerated with distilled 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 reduced 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 antiinflammatory property of the extract was investigated in two animal models of subchronic inflammation (1) in comparison with a number of well established reterence compounds.
One was the air pouch granuloma originally described 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
Introduction
anesthetized with ether and 20 ml of air were injected s.c. into the
Erythraea centauriurn (Gentianaceae) is
cream and the extract at different concentrations in creams were
known in Turkey as "Red Cantarone". The plant has a widespread use in popular medicine against different types of inflamed 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 purpose.
In spite of an extensive search, it was not possible to find any literature concerning the related pharmacological actions of the plant. The present study was, therefore, undertaken to work out whether the aqueous extract 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 Department of the Faculty of Pharmacy of Ege University. After identifica-
tion the samples were dried in the shade at ambient temperature
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
applied topically to the surface of the pouch and spread over a controlled 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 adrenal and thymus weights were also recorded.
The second model used was the polyarthritis, induced 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 approximately 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. Indomethacin 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.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
(25—30°C). The dried plants were roughly cut and ground to pow-
Abstract
Antiinflainmatory. Analgesic, and Antipyretic Effects of an Aqueous Extract ofErythraea centauriurn
Planta Med. 57(1991) 35
Analgesic activity
basic body temperatures. The temperatures of the same animals were recorded 0.5, 1, 2, 3, and 4h after DNP administration. The
The analgesic activity of the extract was assessed
maximum hyperthermia of DNP was observed 0.5 h after its injection, therefore, the extract was administered to the animals in different concentrations i.p. 0.5 h after DNP injection. The rectal body temperatures of the animals were recorded at the same time intervals mentioned above.
in two well established models, the writhing syndrome of the mouse and the hotplate test.
In the first model the writhing syndrome was elicited 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
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 measurement time intervals were as in DNP hyperthermia.
tested in mice (20—30 g) with the hotplate method (53.5°C), 30 and
Results and Discussion Antiinflammatory effects
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 accepted as end-point. The cut- off time was 60 sec (6).
Antipyretic activity The antipyretic effect of the extract was tested in a standard screening for compounds exhibiting this activity. Male, Swiss Albino rats weighing 200—2 50 g were utilized in all experiments and the rectal body temperatures were measured using an applicator type Ri by means of an Electrical Universal Thermometer of the TE3 Ellab type.
In the first test carried out, the body temperatures of the rats which were fasted overnight with water ad lib. were taken initially at time zero. The animals were weighed and injected s.c. in the neck region with either of the following: a 15% solution of brewer's yeast or 0.9% NaC1 solution (10 mi/kg) (7). Temperatures were again taken 2.5 h after the injection of yeast. The initial readings and the 2.5 h readings were combined to form the initial 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 concentration 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
Treatment
N
Daily Cream Dose
Average Exudate
ml S.D.
clearly indicate that 10% extract cream at the dose of 200 mg/day exhibits a significant decrease in exudate formation. 2.5% and 5% extract creams inhibited the exudate formation by aproximately 19% and 42%, respectively. The results of the adjuvant induced polyarthritis 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 evident that the extract was effective in reducing the edematous 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 disease.
% Rel. Organ Weights A B. W} Inhib. mg/loOgbodyweight (g)
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
0.020
0.160
+ 18
Clobetasol 17-butyrate (0.05%) cream
6
50
8.6 0.46"
52
0.016
0.075
+6
Diethylaminesalicylate (4 g/40 g( cream
6
200
4.8 0.39"
73
0.020
0.105
+ 12
Extractcream(l%)
7
200
15.1
16
0.019
0.139
+24
Extractcream(2.5%(
6
200
0.021
0.143
+ 19
7
200
42
0.020
0.113
+ 19
Extractcream(10%(
6
200
14.7 0.58" 10.5 0.61" 9.5 0.33"
19
Extract cream (5 %)
47
0.022
0.110
+ 16
Extract (pure)
6
200
17.4 0.92
3
0.021
0.156
+ 28
0.73"
0.51'
56
a No. of rats.
Change in body weight from initial weight.
** Difference (P < 0.01) from controls;' = difference (P < 0.05( from controls.
Table 1 Effects of the extract and drugs on the air-pouch granuloma.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
The results of the air-pouch granuloma bioassay procedure are given in Table 1. In spite of the advantages and the disadvantages of the method for evaluating topically applied antiinflammatory preparations as discussed by DiPasquale et al. (3) and Fisher (10) our findings
Planta Med. 57(1991)
Tayfun Berkan eta!. Table 2 Effects of the extract and two known an-
Treatment
Average Thickness of the Injected
Dose
% Inhibition of Thickening
Foot
mm±S.D. Control
—
5.8
2.0
Extract
1 mg/day
1.810 0.34" 2.608 0.22" 5.988 0.03
2.4
6.7
Extract
5mg/day
5.900 006'
3.9
6.7
Extract
10mg/day
8.3
12.5
5.2'
Extract
50mg/day
5.632 0.04" 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"
Indomethacin
1 mg/kg/day
Dikiofenac-Na
4mg/kg/day
tiinflammatory drugs on rats with adjuvant disease.
Weight Gains
g±S.D.
6.138 0.20
—
Average Body
70.5
28.0
5.7"
57.5
17.0
2.7" 2.5 2.5
Difference (p
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 Address for correspondence
2
Received: November 10, 1989
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 extract exhibited antiinflammatory and antipyretic activity although no analgesic activity was observed.
Key words Erythraea centauriurn, Gentianaceae, antiinflammatory effects, analgesic effects, antipyretic effects.
der. The weighed powder (230 g) was macerated with distilled 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 reduced 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 antiinflammatory property of the extract was investigated in two animal models of subchronic inflammation (1) in comparison with a number of well established reterence compounds.
One was the air pouch granuloma originally described 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
Introduction
anesthetized with ether and 20 ml of air were injected s.c. into the
Erythraea centauriurn (Gentianaceae) is
cream and the extract at different concentrations in creams were
known in Turkey as "Red Cantarone". The plant has a widespread use in popular medicine against different types of inflamed 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 purpose.
In spite of an extensive search, it was not possible to find any literature concerning the related pharmacological actions of the plant. The present study was, therefore, undertaken to work out whether the aqueous extract 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 Department of the Faculty of Pharmacy of Ege University. After identifica-
tion the samples were dried in the shade at ambient temperature
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
applied topically to the surface of the pouch and spread over a controlled 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 adrenal and thymus weights were also recorded.
The second model used was the polyarthritis, induced 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 approximately 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. Indomethacin 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.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
(25—30°C). The dried plants were roughly cut and ground to pow-
Abstract
Antiinflainmatory. Analgesic, and Antipyretic Effects of an Aqueous Extract ofErythraea centauriurn
Planta Med. 57(1991) 35
Analgesic activity
basic body temperatures. The temperatures of the same animals were recorded 0.5, 1, 2, 3, and 4h after DNP administration. The
The analgesic activity of the extract was assessed
maximum hyperthermia of DNP was observed 0.5 h after its injection, therefore, the extract was administered to the animals in different concentrations i.p. 0.5 h after DNP injection. The rectal body temperatures of the animals were recorded at the same time intervals mentioned above.
in two well established models, the writhing syndrome of the mouse and the hotplate test.
In the first model the writhing syndrome was elicited 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
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 measurement time intervals were as in DNP hyperthermia.
tested in mice (20—30 g) with the hotplate method (53.5°C), 30 and
Results and Discussion Antiinflammatory effects
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 accepted as end-point. The cut- off time was 60 sec (6).
Antipyretic activity The antipyretic effect of the extract was tested in a standard screening for compounds exhibiting this activity. Male, Swiss Albino rats weighing 200—2 50 g were utilized in all experiments and the rectal body temperatures were measured using an applicator type Ri by means of an Electrical Universal Thermometer of the TE3 Ellab type.
In the first test carried out, the body temperatures of the rats which were fasted overnight with water ad lib. were taken initially at time zero. The animals were weighed and injected s.c. in the neck region with either of the following: a 15% solution of brewer's yeast or 0.9% NaC1 solution (10 mi/kg) (7). Temperatures were again taken 2.5 h after the injection of yeast. The initial readings and the 2.5 h readings were combined to form the initial 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 concentration 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
Treatment
N
Daily Cream Dose
Average Exudate
ml S.D.
clearly indicate that 10% extract cream at the dose of 200 mg/day exhibits a significant decrease in exudate formation. 2.5% and 5% extract creams inhibited the exudate formation by aproximately 19% and 42%, respectively. The results of the adjuvant induced polyarthritis 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 evident that the extract was effective in reducing the edematous 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 disease.
% Rel. Organ Weights A B. W} Inhib. mg/loOgbodyweight (g)
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
0.020
0.160
+ 18
Clobetasol 17-butyrate (0.05%) cream
6
50
8.6 0.46"
52
0.016
0.075
+6
Diethylaminesalicylate (4 g/40 g( cream
6
200
4.8 0.39"
73
0.020
0.105
+ 12
Extractcream(l%)
7
200
15.1
16
0.019
0.139
+24
Extractcream(2.5%(
6
200
0.021
0.143
+ 19
7
200
42
0.020
0.113
+ 19
Extractcream(10%(
6
200
14.7 0.58" 10.5 0.61" 9.5 0.33"
19
Extract cream (5 %)
47
0.022
0.110
+ 16
Extract (pure)
6
200
17.4 0.92
3
0.021
0.156
+ 28
0.73"
0.51'
56
a No. of rats.
Change in body weight from initial weight.
** Difference (P < 0.01) from controls;' = difference (P < 0.05( from controls.
Table 1 Effects of the extract and drugs on the air-pouch granuloma.
Downloaded by: Chinese University of Hong Kong. Copyrighted material.
The results of the air-pouch granuloma bioassay procedure are given in Table 1. In spite of the advantages and the disadvantages of the method for evaluating topically applied antiinflammatory preparations as discussed by DiPasquale et al. (3) and Fisher (10) our findings
Planta Med. 57(1991)
Tayfun Berkan eta!. Table 2 Effects of the extract and two known an-
Treatment
Average Thickness of the Injected
Dose
% Inhibition of Thickening
Foot
mm±S.D. Control
—
5.8
2.0
Extract
1 mg/day
1.810 0.34" 2.608 0.22" 5.988 0.03
2.4
6.7
Extract
5mg/day
5.900 006'
3.9
6.7
Extract
10mg/day
8.3
12.5
5.2'
Extract
50mg/day
5.632 0.04" 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"
Indomethacin
1 mg/kg/day
Dikiofenac-Na
4mg/kg/day
tiinflammatory drugs on rats with adjuvant disease.
Weight Gains
g±S.D.
6.138 0.20
—
Average Body
70.5
28.0
5.7"
57.5
17.0
2.7" 2.5 2.5
Difference (p
