Pharmaceutical Biology

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Toxicity Evaluation of Cucurbita maxima. Seed

Extract in Mice

R.C.B. Cruz, C.D. Meurer, E.J. Silva, C. Schaefer, A.R.S. Santos, A. Bella Cruz &
V. Cechinel Filho

To cite this article: R.C.B. Cruz, C.D. Meurer, E.J. Silva, C. Schaefer, A.R.S. Santos, A. Bella
Cruz & V. Cechinel Filho (2006) Toxicity Evaluation of Cucurbita maxima. Seed Extract in Mice,
Pharmaceutical Biology, 44:4, 301-303, DOI: 10.1080/13880200600715886

To link to this article: https://doi.org/10.1080/13880200600715886

Published online: 07 Oct 2008.

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Pharmaceutical Biology
2006, Vol. 44, No. 4, pp. 301–303

Toxicity Evaluation of Cucurbita maxima Seed Extract in Mice

R.C.B. Cruz1, C.D. Meurer1, E.J. Silva1, C. Schaefer1, A.R.S. Santos2, A. Bella Cruz1, and V. Cechinel Filho1
1
Núcleo de Investigações Quı́mico-Farmacêuticas, Centro de Ciências da Saúde, Universidade do Vale do Itajaı́,
Itajaı́, SC, Brazil; 2Departamento de Ciências Fisiol

ogicas, Centro de Ciências Biol

ogicas, Universidade Federal
de Santa Catarina, Florian
opolis, SC, Brazil

Abstract
Pumpkin seeds of Cucurbita maxima Duch. (Cucurbita-
ceae) are frequently used for stomach pain, as an anti-
inflammatory and antipyretic, and for the treatment of
worms. This study was carried out to determine the poss-
ible acute and subacute toxicity effect of the hydroalco-
hol extract from seeds of C. maxima after oral
administration to mice and to identify the target organs
of its possible toxic effects. The results showed that its
average lethal dose (DL50) is higher than 5000 mg=kg
and subacute treatment shows increases in body weight.
The levels of serum alkaline phosphatase, alanine amino-
transferase (ALT), and aspartate aminotransferase
(AST) and the hematological parameters did not show
any alterations. The results suggest that the hydroalco-
holic extract from C. maxima seeds, at a dose of
5000 mg=kg, presents a considerable safety margin, being
devoid of acute toxicity.
Keywords: Acute toxicity, Cucurbita maxima, subacute
toxicity
Introduction
Cucurbita sp. is a vegetable belonging to the family
Curcubitaceae and has been used as a popular vegetable
in cooking since antiquity. Pumpkin seeds are also com-
monly used as a traditional medicine in several countries
for the treatment of ascariasis (Srivastava & Singh, 1967;
Giove Nakazawa, 1996), cestodiasis (Chung & Ko, 1976;
Yang et al., 1994; Mohamoud & El Alfy, 2003), and
schistosomiasis (Chou & Ming, 1960). Pharmacological
studies have also demonstrated antitumor activities
(Xia et al., 2003).
In Brazil, Cucurbita maxima Duch., is popularly
known as pumpkin-moranga or jerimum, where it is
cultivated annually (Correa, 1926). It is yellowish-white
in color and is commonly used in folk medicine to treat
stomach pain and as an anti-inflammatory and antipar-
asitic (Vanaclocha & Ca~ nigueral, 2003). It is also used
as a traditional intestinal antiparasitic in rural and urban
areas (Srivastava & Singh, 1967). Previous pharmaco-
logical studies have demonstrated that this plant presents
strong antimalarial activity (Amorim et al., 1991) and
anthelmintic activity (Amorim & Borba, 1993; Diaz
Obregon et al., 2004).
To the best of our knowledge, only one report exists in
the literature concerning the possible toxic effect of
C. maxima (Queiroz-Neto et al., 1994). This study aimed
to evaluate the toxicity of the hydroalcoholic extract
from C. maxima in order to define the limits of safety
regarding the medicinal use of its seeds.
Materials and Methods
Plant extract
Pumpkin-moranga (Cucurbita maxima Duch.) were
authenticated by MSc. Rene Artur Ferreira (Curso de
Farm
acia, Univali, Itajaı́, SC, Brazil). A voucher speci-
men was deposited in the Barbosa Rodrigues Her-
barium (BRH, Itajaı́, SC, Brazil) under number VC
Filho 060. A pumpkin seed hydroalcoholic extract
was prepared by triturating air-dried seeds and macer-
ating with ethanol 50% at room temperature for 15
days. The solvent was then removed under reduced
pressure.

Accepted: 9 March 2006

Address correspondence to: Alexandre Bella Cruz, UNIVALI=CCS=FARMÁCIA, Rua Uruguai, 458, 88.302-202, Itajaı́, SC, Brazil.
E-mail: bellacruz@univali.br

DOI: 10.1080/13880200600715886 # 2006 Taylor & Francis Group, LLC

302 R.C.B. Cruz et al.

Animals Results and Discussion

Female and male Swiss mice, 25–35 g, from the animal Oral administration of hydroalcohol extract from C.
laboratory of Univali, were kept in a temperature-con- maxima seeds in mice, at doses from 100 to 5000 mg=kg,
trolled environment (25  2C) with a 12-h light-dark did not produce any significant changes in behavior,
cycle. Food and water were freely available for the acute breathing, cutaneous effects, sensory nervous system
toxicity experiment. The same occurred for the subacute responses, or gastrointestinal effects. During the 24-h
toxicity experiment, with food and water being recorded experimental period, no deaths occurred in any of the
every 2 days. groups. These results indicate that DL50 of hydroalco-
holic extract of C. maxima is higher than 5000 mg=kg
and presented a considerable safety margin, being appar-
Acute toxicity ently devoid of acute toxicity for male and female mice.
The mice were randomly divided into four experimental The results show that the body weight of the animals
groups and a control group, with 10 animals in each treated with the 1000 mg=kg dose during the experi-
group, and kept in propylene boxes. The control group mental 30-day period presented significant changes com-
received saline, and each of the experimental groups pared with the control group (Table 1). These results can
received hydroalcoholic extract at doses of 100, 500, be justified by the chemical composition and favorable
1000, and 5000 mg=kg, administered through an orogas- nutritional value of the pumpkin seed (Vanaclocha &
tric tube. All the animals were kept under constant obser- Ca~nigueral, 2003; Achu et al., 2005). Zdunczyk and
vation for 3 h, and for 24 h after administering the co-workers (1999) have reported that the nutritional
extract, to observe any changes in general behavior or value of pumpkin seed has a higher protein efficiency
other physiological activities. At the end of the ratio (PER) value than the soybean meal diet.
experiment, each animal was sacrificed by cervical The macroscopic analysis of the target organs of the
displacement. treated animals (liver, heart, lung, spleen, and left kid-
ney) did not show any significant changes (color, texture)
when compared with the control group (Table 2).
Subacute toxicity The hematology parameters, red blood cell, hemato-
The mice were divided at random into an experimental crit, leukocyte, and differential of cells, did not show
group and a control group, with 10 animals in each.
The control group received saline, and the experimental Table 1. Variation of the mean weight of the mice with treat-
group received hydroalcoholic extract (1000 mg=kg), ment (oral route) for 30 days, with Cucurbita maxima seed
extract at a dose of 1000 mg=kg.
administered orally through an orogastric tube for 30
days. The animals were weighed every 2 days. At the Time (day) Control 1000 mg=kg
end of the experimental period, blood was collected by
orbital sinus venipuncture, under ether anesthesia, for 1 30.75  3.15 28.01  2.68
biochemical and hematological analysis. After the blood 5 31.72  3.22 35.62  4.50
10 33.02  3.75 39.00  5.17
collection, the animals were sacrificed by cervical dis-
15 35.61  4.10 41.62  5.33
placement, and the target organs (liver, heart, spleen, left 20 35.70  4.57 43.42  4.43
kidney, and left lung) were removed for macroscopic 25 37.50  4.50 45.52  4.73
analysis. 30 39.37  4.72 48.01  3.74
The biochemical parameters evaluated included assays
for serum alkaline phosphatase (Alk. Phosph.), alanine Mean values of 10 animals  SD. p < 0.05.
aminotransferase (ALT), aspartate aminotransferase
(AST), creatinine, and urea using commercial kits Table 2. Effect of Cucurbita maxima seed extract on body and
(LABTEST, Lagoa Santa, MG, Brazil). The hematology organ weight.
parameters were determined for the control and
Parameter Control 1000 mg=kg
1000 mg=kg group and included a red blood cell count,
hematocrit and leukocyte count. Body (g) 39.37  4.72 48.01  3.74
Liver (mg) 1812.50  106.25 2550.00  168.75
Heart (mg) 185.93  4.69 214.06  7.81
Statistical methods Left lung (mg) 215.62  6.25 243.75  12.50
The results are presented as mean  SD, and the statisti- Spleen (mg) 145.83  15.83 175.00  16.66
Left kidney (mg) 208.33  8.33 307.50  14.16
cal significance between the groups was analyzed by
means of analysis of variance, followed by Dunnett’s Mean values of 10 animals  SD.  p < 0.05;  p < 0.01 vs. con-
multiple comparison test. P values of less than 0.05 were trol group (Dunnett’s test). Treatment for 30 days. The control
considered as indicative of significance. group received saline.
 Toxicity evaluation of Cucurbita maxima seed 303

Table 3. Blood profile after treatment with the Cucurbita max- seeds from different regions in Cameroon. Afr J Bio-
ima seed hydroalcoholic extract. technol 4: 1329–1334.
Amorim CZ, Marques AD, Cordeiro RS (1991): Screening
Parameter Control 1000 mg=kg
of the antimalarial activity of plants of the Cucurbita-
Hemoglobin (mg=dl) 13.67  0.44 13.82  0.23 ceae family. Mem Inst Oswaldo Cruz 86(Suppl 2):
Red blood cell (mm3) 8.53  0.43 8.08  0.20 177–180.
Hematocrit (%) 42.88  1.17 44.83  0.29 Amorim A, Borba HR (1993): Ação anti-helmı́ntica de
Leukocyte (106=ml) 8.43  0.68 6.76  0.69 plantas VII - Triagem de 14 extratos brutos sobre
Mean values of 10 animals  SD. p < 0.05 vs. control group Vampirolepsis nana em camundongos. Rev Bras Farm
(Dunnett’s test). 74: 2–3.
Correa MP (1926): Dicion
ario das Plantas Úteis do Brasil e
das Ex
oticas Cultivadas, Vol I. Ministério da Agricul-
Table 4. Effect of Cucurbita maxima seed extract on biochemi- tura. Rio de Janeiro, pp. 6, 7, 10, 351.
cal parameters.
Chou HC, Ming H (1960). Pumpkin seed (Cucurbita
Parameter Control 1000 mg=kg moschata) in the treatment of acute schistosomiasis.
Chin Med J 80: 115–120.
Alkaline phosphatase (U=l) 204.83  15.63 224.37  24.00 Chung WC, Ko BC (1976). Treatment of Taenia saginata
ALT (U=l) 32.54  3.05 46.06  5.09 infection with mixture of areca nuts and pumpkin
AST (U=l) 111.86  5.09 103.72  10.17
seeds. Zhonghua Min Guo Wei Sheng Wu Xue Za Zhi
Creatinine (mg=dl) 0.216  0.03 0.283  0.03
9: 31–35.
Urea (mg=dl) 40.00  3.75 56.25  2.50
Diaz Obregon D, Lloja Lozano L, Carbajal Zuniga V
Mean values of 10 animals  SD. p < 0.05 vs. control group (2004). Preclinical studies of Cucurbita maxima (pump-
(Dunnett’s test). kin seeds) a traditional intestinal antiparasitic in rural
urban areas. Rev Gastroenterol Peru 24: 323–327.
any considerable alterations in the animals that received Giove Nakazawa RA (1996). Traditional medicine in the
the hydroalcohol extract compared with the control treatment of enteroparasitosis. Rev Gastroenterol Peru
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gest that the hydroalcoholic extract from C. maxima Mohamoud LH, El Alfy NM (2003). Effect of some chemi-
seeds, at a dose of 1000 mg=kg, does not interfere in cal agents on the viability of Cysticercus bovis. J Egypt
the level of production=inhibition of red blood cells. Soc Parasitol 33: 391–396.
Table 4 shows the results of the biochemical assays Queiroz-Neto A, Mataqueiro MI, Santana AE, Alessi AC
with the serum, for mice treated, or not treated, with a (1994). Toxicologic evaluation of acute and subacute
hydroalcoholic extract from C. maxima seeds. The serum oral administration of Cucurbita maxima seed extracts
evaluation of the enzymes Alk. Phosph., AST, and ALT to rats and swine. J Ethnopharmacol 43: 45–51.
is an important reference for the evaluation of hepato- Srivastava MC, Singh SW (1967). Anthelmintic activity of
toxicity, and these results do not demonstrate any signifi- Cucurbita maxima (kaddu) seeds. Indian J Med Res
cant alteration. However, we stress that the creatinine 55: 629–632.
and urea levels (Table 4) presented abnormal alterations Vanaclocha B, Ca~ nigueral S (2003): Fitoterapia, Vademécum
when compared with the control group, which can be de Prescripción, 4 ed., Barcelona: Masson, pp. 153–154.
related to the protection of the catabolism or high pro- Yang X, Xu H, Cao Z, Wang W (1994). Establishment of
tein diet, therefore the urinary flow of the mice was con- mouse model for Cysticercosis cellulosae. Zhongguo Ji
sidered normal. Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 12:
Our results showed that hydroalcoholic extract from 297–299.
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mice. These results reinforce the findings for this pump- some-inactivating protein from the mature seeds of
kin against rats and swine (Queiroz Neto et al., 1994). pumpkin (Cucurbita moschata), and preparation of its
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Zdunczyk Z, Minakowski D, Frejnagel S, Flis M (1999):
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