J Pharm Pharmaceut Sci (www.cspsCanada.

org) 8(2):370-373, 2005

Acute and subacute toxicity of the trypanosomicidal activity (0) and smooth muscle
hydroalcoholic extract from Wedelia relaxant effect (0). Due to the widespread use of this
paludosa (Acmela brasiliensis) plant by the rural communities to treat several
diseases, the objective of the present study was to
(Asteraceae) in mice
obtain data on the safety of the crude extract. The
Cristiani Bürger, Doris Raquel Fischer, Dórys Angela acute and subacute oral toxicity of the
Cordenunzzi, Anna Paula de Borba Batschauer, hydroalcoholic extract from aerial parts of this plant
Valdir Cechinel Filho, Adair Roberto dos Santos in mice was assessed. The changes in selected
Soares biochemical and hematological parameters were also
determined.
Núcleo de Investigações Químico-Farmacêuticas
(NIQFAR), Centro de Ciências da Saúde (CCS),
Universidade do Vale do Itajaí (UNIVALI), Itajaí, SC, Brazil
METHODOLOGY

Received June 6, 2005, Revised June 25, 2005, Accepted June Plant material
27, 2005, Published August 19, 2005 The aerial parts of Wedelia paludosa DC were
collected next to NIQFAR/UNIVALI, in the town of
Abstract PURPOSE. The present study was carried Itajaí, in the State of Santa Catarina. The plant
out to evaluated acute and subacute toxicity of a material was authenticated by Dr. Ademir Reis
hydroalcoholic extract from aerial parts of Wedelia (Department of Botany, UFSC, Florianópolis) and a
paludosa (Asteraceae). METHODS. Toxicity of W. voucher specimen was deposited at the Barbosa
paludosa was evaluated in Swiss mice after Rodrigues Herbarium (Itajaí), under number VC
ingestions of the extract during one day (acute Filho 002. The aerial parts of the plant were air-
model) and during 15 days (subacute model). dried, cut into small pieces and macerated with 50 %
RESULTS. The results showed that the LD50 of the ethanol (w/w) at room temperature for 15 days. After
extract is higher than 4000 mg/kg and the subacute filtration, the solvent was removed under reduced
treatment did not shows any change in corporal pressure and the hydroalcoholic extract was then
weight and hematological parameters. However, a obtained.
change in liver weight but not in hepatic enzymes
was observed. This suggests that the liver function is Animals
not altered by Wedelia paludosa in this study. Some Acute toxicity
changes in the creatinine content were observed, but The toxicity study as carried out using female and
could not be related with the extract dose. male Swiss mice (25-35 g). Animals were kept in a
CONCLUSIONS. The results suggest that the plant temperature-controlled environment (23 ± 2ºC) with
seems to be destituted of toxic effects in mice. a 12 h light-dark cycle and food and water were
freely available. Ethics Committee of UNIVALI
INTRODUCTION approved the protocol for these experiments under
number 314/2004. The animals were divided into
Wedelia paludosa DC, recently reclassified as one control group and five treated groups, each
Acmela brasiliensis (Asteraceae), is an ornamental group consisting of ten animals. The control group
plant, being widely used in natural medicine in South received saline and each treated group received the
Brazil. This plant is used for the treatment of several hydroalcoholic extract in a dose of 100, 500, 1000,
ailments, including respiratory infections and pain 2000 and 4000 mg/kg by gavage. These doses were
(0, 0). Our research group has previously choose because were 10-100 times higher than
investigated the phytochemical and pharmacological effective doses in other studies. The animals were
properties of this plant. In this context, its analgesic observed continuously for 3 h ,and then they were
(0, 0), antimicrobial and antidiabetic effects (0), were observed each hour during 24 h after administering
described and related with the presence of terpenes the extract to observe any changes in general
and flavonoids, including kaurenoic acid and luteolin behavior or other physiological activities. At the end
(0, 0, 0). Other authors showed that kaurane of the experiment animals were sacrificed by cervical
diterpenes from W. paludosa display displacement.
Corresponding author: Cristiani Bürger: Núcleo de
Investigações Químico-Farmacêuticas (NIQFAR), CCS,
Universidade do Vale do Itajaí (UNIVALI). R. Uruguai, 458, CP
360, 88302-202, Itajaí, SC, Brazil. [email protected]

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Subacute toxicity observed during the experimental period (24 h).

Female Swiss mice, 25-35 g, were kept in a During 24 h of the experiment, no deaths occurred in
temperature-controlled environment (23 ± 2ºC) with any of the groups. These results showed that in
a 12 h light-dark cycle. Food and water were freely single dose, there are no adverse effects of W.
available and were recorded each 3 days. The paludosa, indicating that the medium lethal dose
animals were divided into one control group and four (LD50) is higher than 4000 mg/kg for male and
treated groups (500, 1000, 2000 and 4000 mg/kg), female mice.
each group consisting of ten animals. The control The results showed that a hydroalcoholic
group received saline and each treated group extract from Wedelia paludosa is safe in oral
received the hydroalcoholic extract by gavage for 15 administration in rodents. The doses used in this
days (once a day) . The animals were weighed each 3 study were 10 - 100 times higher than those used in
days. At the end of the experiment, blood was other experimental pharmacological studies, such as:
collected from the orbital sinus under ether decrease of blood glucose levels in alloxan-induced
anesthesia for biochemical and hematological diabetic rats (300 mg/kg) (0); inhibition of pain
analysis. After the blood collection, the animals were caused by intraperitoneal administration of extract on
sacrificed by cervical displacement and selected writing test in mice (1mg/kg of roots extract and 3
organs (liver, heart, spleen, left kidney and left lung) mg/kg of leaves and stem extracts (0).
were removed for macroscopic analysis. The The treatment with the extract did not
biochemical parameters evaluated included decrease the water and food consumption (data not
creatinine, calcium, inorganic phosphorus, alanine show). The body weight (table 1) of the animals
aminotransferase (ALT), aspartate aminotransferase treated with hydroalcoholic extract once a day during
(AST), gamma-glutamyl transferase and serum 15 days (subacute treatment) did not show any
alkaline phosphatase and were assessed using significant change when compared with the control
commercial kits. The hematology parameters were group, although had a tendency to decrease body
determined for the control and 1000 mg/kg groups weight (2000 and 4000 mg/kg). This decrease can be
and included red blood-cell count, hematocrit and associate with the decrease of liver weight at the
leukocyte count. doses of 2000 and 4000 mg/kg in comparison with
the control group without any concomitant alteration
Statistical analysis in the activity of alanine aminotransferase, aspartate
The results are presented as mean ± s.d. and the aminotransferase, and gama-glutammyl transferase.
statistical significance between the groups was Estimation of the serum activity gamma-glutammyl
analyzed by means of an analysis of variance transferase, alanine aminotransferase and aspartate
followed by Dunnett's multiple comparison test. P aminotransferase is one of the most widely used
values less than 0.05 were considered as significant. means of measuring hepatocellular injury (0). The
macroscopic analysis of the target organs of the
RESULTS AND DISCUSSION treated animals (liver, lung, heart, spleen and left
kidney) did not show significant changes in color
The use of herbal preparations as in the treatment of and texture when compared with the control group.
diseases is very common in the rural communities of The results of organs weight are summarized in table
Brazil. Wedelia paludosa is frequently used for the 1.
treatment of infections, pathologies of the respiratory During the experimental period, there were
tract, fever and others diseases involving no treatment-related effects on the hematological
inflammation and pain (0, 0). The importance of this parameters evaluated (table 2). On the other hand,
plant in folk medicine as well as its promising alterations in the serum concentration of creatinine
pharmacological properties verified in our were observed (table 3). As the macroscopic
laboratories (0, 0, 0, 0, 0), make studies about its appearance and weight of the kidney was not altered,
toxicity very important. hence, the possibility of renal injuries could not be
Oral administration of the hydroalcoholic confirmed.
extract of W. paludosa in doses from 100 to 4000
mg/kg did not produce significant changes in
behavior, breathing, cutaneous effects, sensory
nervous system responses, and gastrointestinal
effects in male and female mice. These effects are

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Table1. Effect of oral administration of W. paludosa extract on body and organs weight.
Dose (mg/kg): Control 500 1000 2000 4000
Body (g) 34.1 ± 3.17 34.7 ± 1.90 36.0 ± 3.77 32.0 ± 3.8 33.0 ± 2.77
Liver (g) 1.559 ± 0.200 1.415 ± 0.140 1.468 ± 0.225 1.3 ± 0.177 *
1.12 ± 0.176**
Heart (g) 0.144 ± 0.018 0.151 ± 0.017 0.163 ± 0.037 0.155 ± 0.028 0.12 ± 0.028
Left lung (g) 0.213 ± 0.037 0.191 ± 0.016 0.235 ± 0.040 0.242 ± 0.064 0.18 ± 0.040
Spleen (g) 0.186 ± 0.092 0.156 ± 0.031 0.165 ± 0.051 0.126 ± 0.013 0.14 ± 0.0296
Left kidney (g) 0.165 ± 0.021 0.157 ± 0.017 0.175 ± 0.025 0.171 ± 0.027 0.14 ± 0.0237
Mean values of 10 animals ± S.D. p<0.05; p< 0.01 vs. control group (Dunnett's test). Control group received saline. No
* **

significant difference was observed in any parameter, except in liver (2000 and 4000 mg/kg).

Table 2. Hematological parameters after 15 days treatment with the W. paludosa extract.
Parameter Control 1000 mg/kg 4000 mg/kg
Red blood cell (mm3) 9.043 ± 0.370 8.095 ± 0.451 8.81 ± 0.129
Hematocrit (%) 46.1 ± 4.63 44.1 ± 1.524 47.049 ± 2.63
Leukocyte (x106/mL) 7.550 ± 2.192 8.730 ±2.493 7.14 ± 1.689
Values are mean of 10 animals ± S.D. (Dunnett's test).
No significant difference was observed in any parameter.

Table 3. Effect of treatment with W. paludosa extract on biochemical parameters.

Dose (mg/kg) Control (n=9) 500 1000 2000 4000
(n=10) (n=9) (n=8) (n=7)
Creatinine (mg/dL) 0.259 ± 0.050 0.285 ± 0.04 0.159 ± 0.04** 0.215 ± 0.06 0.248 ± 0.025
Calcium (mg/dL) 7.489 ± 0.528 7.976 ± 0.91 7.754 ± 0.322 8.154 ± 0.382 8.317± 0.807
Inorganic phosphorus (mg/dL) 6.650 ± 0.547 6.210 ± 0.62 6.770 ± 0.641 6.410 ± 1.162 6.836 ± 0.671
ALT (U/L) 111.9 ± 21.20 113.9 ± 10.5 91.99 ± 19.89 98.29 ± 31.77 90.51± 15.25
AST (U/L) 54.65 ± 10.15 51.57 ± 8.61 52.38 ± 8.072 48.88 ± 8.059 53.67 ± 12.44
Gamma-glutamyl transferase (U/L) 2.133 ± 1.920 2.780 ± 1.83 2.03 ± 1.481 2.401 ± 2.81 3.42 ± 1.616
Values are mean ± S.D. p< 0.01 vs. control group (Dunnett's test).
**

ALT – alanine aminotransferase

AST – aspartate aminotransferase

Administration of hydroalcoholic extract of In summary, our study demonstrated that W.

W. paludosa in mice not induced any changes in paludosa seems to be destitute of toxic effects, which
these enzymes, although the liver weight decreased. could be compromise the medicinal use of this plant
This observation could indicate that liver function is in folk medicine. However, further studies are
preserved by oral administration of W. paludosa. Its necessary, such as histological and morphological
important to mention that studies on other species of experiments, to confirm this evidence.
the genus Wedelia showed that: i) W. calendulacea
has protective activity against liver injury in vivo (0); ACKNOWLEDGEMENTS
ii) W. glauca caused liver necrosis in rats (0), sheep
and cattle (0); iii) W. chinensis has an The authors are grateful to CNPq (Brazil) for
hepatoprotective effect against liver injury (0). financial support.

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