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Effect of essential oil from Citrus aurantium in maternal reproductive outcome and fetal anomaly frequency in rats

Abstracts

Citrus aurantium L., commonly known as bitter orange, is widely used in folk medicine, but there is little data in the literature about the effects on pregnancy. The aim of the present study was to evaluate the influence of essential oil obtained from fruits of Citrus aurantium on the maternal reproductive outcome and fetal anomaly incidence in rats. Pregnant Wistar rats were randomized into four groups (n minimum = 12 animals/group): G1 = control, G2 to G4 = treated with essential oil from C. aurantium at dose 125, 250 and 500 mg/kg, respectively. Rats were orally treated, by gavage, with plant essential oil or vehicle during pre-implantation and organogenic period (gestational day 0-14). On gestational day 20 the rats were anaesthetized and the gravid uterus was weighed with its contents and the fetuses were analyzed. Results showed that the treated group with 500 mg/kg presented decreased placental weights and placental index, although the treatment with bitter orange essential oil did not show any alteration in maternal reproductive performance, toxicological effect, changes in ossification sites, and malformation index. In conclusion, the treatment of Citrus aurantium essential oil was not teratogenic and did not alter the maternal reproductive outcome.

Citrus aurantium; essential oil; limonene; malformation; pregnancy; reproductive outcome


Citrus aurantium L., conhecida popularmente como laranja amarga, é amplamente utilizada na medicina popular, mas há poucos dados na literatura sobre seus efeitos na gestação. O objetivo do presente estudo foi avaliar a influencia do óleo essencial obtido das frutas de Citrus aurantium no desempenho reprodutivo materno e na incidência de anomalias fetais em ratos. Ratas Wistar prenhes foram randomizadas em quarto grupos (n mínimo = 12 animais/grupo): G1 = controle, G2 à G4 = tratados com óleo essencial de C. aurantium nas doses de 125, 250 e 500 mg/kg, respectivamente. Ratas foram tratadas oralmente, por gavage, com óleo essencial da planta ou veículo durante os períodos de pré-implantação e organogênese (dias de prenhez 0-14). No dia 20 de prenhez as ratas foram anestesiadas e o útero foi pesado com seu conteúdo e os fetos foram analisados. Resultados mostraram que o grupo tratado com 500 mg/kg apresentou diminuição do peso e índice placentário, embora o tratamento com óleo essencial de laranja amarga não mostrou nenhuma alteração no desempenho reprodutivo materno, efeito tóxico, mudanças na ossificação e nas taxas de malformações. Concluindo, o tratamento com óleo essencial de Citrus aurantium não foi teratogênico e não alterou o desempenho reprodutivo materno.

Citrus aurantium; óleo essencial; limo neno; malformação; prenhez; desempenho reprodutivo


INTRODUCTION

A large number of plants have been used in folk medicine for centuries. It is common to assume that medicinal plants of traditional use have already been tested and ratified by its prolonged use in the human species. Consequently, they are considered effective medicines and it is presumed that they present no side effects and do not need evaluation. However, medicinal plants do not differ from any another synthetic chemical substance and its use must be based on several experimental evidences, confirming that the possible injury cause by its usage is supplanted by the benefits (Lapa et al. 2004LAPA AJ, SOUCCAR C, LIMA-LANDMAN MTR, GODINHO RO AND LIMA TCM. 2004. Farmacologia e toxicologia de produtos naturais. In: SIMÕES CMO, SCHENKEL EP, GOSMAN G, MELLO JCP, MENTZ LA AND PETROVICK PR (Eds), Farmacognosia: Da planta ao medicamento, Porto Alegre: UFRGS, Porto Alegre, Brasil, p. 247-262.).

Fruits of Citrus aurantium L. (Rutaceae family), commonly known as bitter orange, sour orange or Seville orange, are used as food, but this popular plant is more commonly used as a medicinal or dietary supplement (Fugh-Berman and Myers 2004FUGH-BERMAN A AND MYERS A. 2004. Citrus aurantium, an ingredient of dietary supplements marketed for weight loss: current status of clinical and basic research. Exp Biol Med 229: 698-704.). Similar to ephedra, it contains alkaloids that are adrenergic agonists and is often incorporated into supplements designed to aid in weight loss (Haaz et al. 2006HAAZ S, FONTAINE KR, CUTTER G, LIMDI N, PERUMEAN-CHANEY S AND ALLISON DB. 2006. Citrus aurantium and synephrine alkaloids in the treatment of overweight and obesity: an update. Obes Rev 7: 79-88.). It was confirmed that its essential oil has an antibiotic effect against Helicobacter pylori (Bergonzelli et al. 2003BERGONZELLI GE, DONNICOLA D, PORTA N AND CORTHÉSY-THEULAZ IE. 2003. Essential oils as components of a diet-based approach to management of Helicobacter infection. Antimicrob Agents Ch 47: 3240-3246.). Novel triterpenoid from C. aurantium L. also possesses chemopreventive properties against human colon cancer cells (Gougeon et al. 2005GOUGEON R, HARRIGAN K, TREMBLAY J, HEDREI P, LAMARCHE M AND MORAIS JA. 2005. Increase in the thermic effect of food in women by adrenergic amines extracted from Citrus aurantium. Obes Res 13: 1187-1194.). Given that limonene and essential oil of C. aurantium are excellent flavoring agents and also present gastroprotective actions, they are being considered promising for the development of a new drug for the prevention of gastric damage (Moraes et al. 2009MORAES TM, KUSHIMA H, MOLEIRO FC, SANTOS RC, ROCHA LR, MARQUES MO, VILEGAS W AND HIRUMA-LIMA CA. 2009. Effects of limonene and essential oil from Citrus aurantium on gastric mucosa: role of prostaglandins and gastric mucus secretion. Chem Biol Interact 180: 499-505.). However, safety information is extremely limited, and because Citrus aurantium contains the sympathomimetic drug m-synephrine (phenylephrine), consumption of the herb may lead to increased blood pressure and risk of adverse cardiovascular events (Bent et al. 2004BENT S, PADULA A AND NEUHAUS J. 2004. Safety and efficacy of Citrus aurantium for weight loss. Am J Cardiol 94: 1359-1361., Hansen et al. 2011HANSEN DK, GEORGE NI, WHITE GE, PELLICORE LS, ABDEL-RAHMAN A AND FABRICANT D. 2012. Physiological effects following administration of Citrus aurantium for 28 days in rats. Toxicol Appl Pharmacol 261: 236-247.). Hansen et al. (2012) verified that synephrine, a component in the bitter orange extract, did not produce developmental toxicity in rats when administered in lower dose. Arbo et al. (2009)ARBO MD, SCHMITT GC, LIMBERGER MF, CHARÃO MF, MORO AM, RIBEIRO GL, DALLEGRAVE E, GARCIA SC, LEAL MB AND LIMBERGER RP. 2009. Subchronic toxicity of Citrus aurantium L. (Rutaceae) extract and p-synephrine in mice. Regul Toxicol Pharm 54: 114-117. indicated a low subchronic toxicity and possible alteration in the oxidative metabolism of the C. aurantium and synephrine in mice.

A great number of experimental studies had indicated that the prenatal administration of plants can result in structural and functional suppression of the growth, fetal anomaly and death (Lemonica and Alvarenga 1994LEMONICA IP AND ALVARENGA CM. 1994. Abortive and teratogenic effect of Acanthospermum hispidum DC. and Cajanus cajan (L.) Millps. in pregnant rats. J Ethnopharmacol 43: 39-44., Damasceno and Lemonica 1999DAMASCENO DC, KISS AC, SINZATO YK, CAMPOS KE, RUDGE MV, CALDERON IP AND VOLPATO GT. 2011a. Maternal-fetal outcome, lipid profile and oxidative stress of diabetic rats neonatally exposed to streptozotocin. Exp Clin Endocrinol Diabetes 119: 408-413., Damasceno et al. 2002a, Mello et al. 2005MELLO FB, JACOBUS D, CARVALHO K AND MELLO JRB. 2005. Effects of Lantana camara (Verbenaceae) on general reproductive performance and teratology in rats. Toxicon 45: 459-466., Dallaqua et al. 2013DALLAQUA B, SAITO FH, RODRIGUES T, CALDERON IM, RUDGE MV, VOLPATO GT AND DAMASCENO DC. 2013. Azadirachta indica treatment on the congenital malformations of fetuses from rats. J Ethnopharmacol 150: 1109-1113.). However, several plants had already been studied during pregnancy, and showed no deleterious effects (Monteiro et al. 2001MONTEIRO MHD, GOMES-CARNEIRO MR, FELZENSZWALB I, CHAHOUD I AND PAUMGARTTEN FJR. 2001. Toxicological evaluation of a tea from leaves of Vernonia condensate. J Ethnopharmacol 74: 149-157., Rudge et al. 2007RUDGE MVC, DAMASCENO DC, VOLPATO GT, ALMEIDA FGE, CALDERON IMP AND LEMONICA IP. 2007. Effect of Ginkgo biloba on reproductive outcome and oxidative stress biomarkers of streptozotocin-induced diabetic rats. Braz J Med Biol Res 40: 1095-1099., Volpato et al. 2008VOLPATO GT, CALDERON IM, SINZATO S, CAMPOS KE, RUDGE MV AND DAMASCENO DC. 2011. Effect of Morus nigra aqueous extract treatment on the maternal-fetal outcome, oxidative stress status and lipid profile of streptozotocin-induced diabetic rats. J Ethnopharmacol 138: 691-696., Volpato et al. 2011).

In spite of its popular use, there is little data in the literature about the effects of Citrus aurantium on pregnancy. Therefore, the objective of the current study was to evaluate the effect of essential oil from C. aurantium on the maternal reproductive outcome and fetal anomaly incidence in rats.

MATERIALS AND METHODS

PREPARATION OF ESSENTIAL OIL

A plant sample of the specie Citrus aurantium was collected and the exsiccates deposited in the Herbarium "Irina D. Gemtchujnicov" - BOTU, Department of Botany at UNESP - Univ Estadual Paulista (BOTU 23123). Fruit peels were cut manually into small fragments (approximately 2 mm×10 mm) for better vapor penetration to yield, and in turn to better access to essential oil, which was extracted by hydro distillation in a Moritz apparatus, using approximately 200 g of peels, for 75 min. Extractions were performed in triplicate (Rozza et al. 2011ROZZA AL, MORAES TM, KUSHIMA H, NUNES DS, HIRUMA-LIMA CA AND PELLIZZON CH. 2011. Involvement of glutathione, sulfhydryl compounds, nitric oxide, vasoactive intestinal peptide, and heat-shock protein-70 in the gastroprotective mechanism of Croton cajucara benth. (Euphorbiaceae) essential oil. J Med Food 14: 1011-1017.).

EXPERIMENTAL ANIMALS

Female and male Wistar rats (210-230 g) were obtained from the UNESP breeding center, and were maintained under standard laboratory conditions (22 ± 3°C, humidity 50±10%,12-h light/dark cycle), with pelleted food (Purina rat chow, Purina(r), São Paulo, SP, Brazil) and tap water ad libitum. The animals were cared for in accordance with the Principles of the Guide for Care and Use of Experimental Animals. The local Committee of Ethics in Animal Experimentation approved all experimental proce dures of this study (49/08-CEEA).

PREGNANCY DIAGNOSTICS AND EXPERIMENTAL GROUPS

After acclimation period, the female rats were mated overnight. The morning on which sperm were found in the vaginal smear was designated as gestational day (GD) 0.

The rats were distributed into four experimental groups (n minimum = 12 animals/group): G1 = control (vehicle), G2 to G4 = treated with essential oil from C. aurantium at dose 125, 250 and 500 mg/kg, respectively. Rats were orally treated, by gavage, with essential oil from C. aurantium or vehicle (Tween(r) 80 at 8%, maximum volume of 0.20 mL) during pre-implantation and organogenic period (from GD 0 to GD 14). Previous studies verified that lowest effective dose of oil was 250 mg/kg (Moraes et al. 2013, Polo et al. 2012POLO CM, MORAES TM, PELLIZZON CH, MARQUES MO, ROCHA LR AND HIRUMA-LIMA CA. 2012. Gastric ulcers in middle-aged rats: The healing effect of essential oil from Citrus aurantium L. (Rutaceae). Evid Based Complement Alternat Med 2012: 509451.). The US Environmental Protection Agency suggests that doses be scaled from rats to humans on the basis of the ratio of human body weight to rat body weight raised to the 0.667 power, as follows: Dose human =Dose rat x (body weight human/ body weight rat) 0.667 (Bachmann et al. 1996BACHMANN K, PARDOE D AND WHITE D. 1996. Scaling basic toxicokinetic parameters from rat to man. Environ Health Perspect 104: 400-407.). Thus, the high dose used to treat rats is approximately 19 g/kg for humans.

COURSE OF PREGNANCY

Maternal weight, food intake and water consumption were measured about every 7 days up to the end of pregnancy, at approximately 9 a.m. On GD 20 the rats were anesthetized by sodium pentobarbital and sacrificed by inducing pneumothorax. The gravid uterus was weighed and dissected to count dead and live fetuses, resorption, implantation sites, and corpora lutea numbers. The number of implantation sites was determined by the Salewski method (Salewski 1964SALEWSKI E. 1964. Farbemethode zum markroskopishen nachweis von implantatconsstellen na uterus der ratter naunyn schmuderbergs. Arch Pharmacol 247: 367.). The rate of pre-implantation loss was calculated as: No. of corpora lutea - No. of implantations x 100/ No. of corpora lutea, and post- implantation loss rate was calculated as: No. of implantations - No. of live fetuses x 100/ No. of implantations (Damasceno et al. 2008). The fetuses and placentas were weighed to calculate the placental index: placental weight/fetal weight. The mean birth weight of the control pups (G1) was 4.2 ± 0.4 g. Newborns in the experimental groups whose birth weights did not diverge more than ± 1.0 standard deviation (SD) from the G1 mean (i.e., those that were within the 3.8- to 4.6- g range) were classified as appropriate for pregnancy age (APA). Those whose weights were at least 1.0 SD greater than the G1 mean birth weight were classified as large for pregnancy age (LPA). Those whose birth weights were at least 1.0 SD lower than the G1 mean birth weight were classified as small for pregnancy age (SPA) (Volpato et al. 2009). The fetuses were evaluated on a microscope with respect to incidence of external anomaly. After external analysis, half the fetuses were fixed in Bouin's fluid and serial sections were prepared as described by Wilson (1965)WILSON JG. 1965. Methods for administering agents and detecting malformations in experimental animal. In: WILSON JG AND WARKANY J (Eds), Teratology: Principles and Techniques. Chicago: University of Chicago Press, Chicago, USA, p. 262-277. for visceral examination. The remaining fetuses were prepared for examination of the skeletons by the staining procedure of Staples and Schnell (1964)STAPLES RE AND SCHNELL VL. 1964. Refinements in rapid clearing technique in the KOH-alizarin red S method for fetal bone. Stain Technol 39: 61-63.. The ossification degree was evaluated using the parameters proposed by Aliverti et al. (1979)ALIVERTI V, BONANOMI L, GIAVINI E, LEONE VG AND MARIANI L. 1979. The extent of fetal ossification as an index of delayed development in teratogenic studies on the rat. Teratology 20: 237-242..

STATISTICAL ANALYSES

For comparison of the mean values between the experimental groups, analysis of variance followed by Tukey's test were used. The proportions were calculated by the Fisher Exact test. Differences were considered statistically significant when p< 0.05.

RESULTS

BODY WEIGHT GAIN, WATER INTAKE, AND FOOD CONSUMPTION

As shown in Table I, the treatment with essential oil from Citrus aurantium did not interfere with body weight gain, water intake, and food consumption. These parameters were similar to control group during the entire experimental period (Table I).

Table I
Body weight, water intake and food consumption of rats treated or not with Citrus aurantium essential oil, from day 0 to 14 of pregnancy.

MATERNAL REPRODUCTIVE OUTCOME

The treatment with essential oil did not alter the parameters of maternal reproductive outcome (number of corpora lutea, implantation, resorptions, live and dead fetuses, maternal weight gain, gravid uterus weight, fetal weight, and sex ratio) compared to control group. The G4 group presented decreased placental weights and placental index compared to G1 group (Table II).

Table II
Maternal reproductive outcome of rats treated or not with Citrus aurantium essential oil, from day 0 to 14 of pregnancy.

OSSIFICATION SITES

As shown in Table III, treated groups presented the number of forepaw and hindpaw phalanx, metatarsus, metacarpus, caudal vertebra, sternebrae, and total ossification sites similar to G1 group (Table III).

Table III
Ossification sites of rats treated or not with Citrus aurantium essential oil, from day 0 to 14 of pregnancy.

ANOMALY FREQUENCY

Data on the occurrence of external, skeletal and visceral anomalies indicated that the treatment with essential oil did not modify the total number of fetuses with alterations (Table IV).

Table IV
Anomaly frequency of rats treated or not with Citrus aurantium essential oil (500 mg/kg body weight/day), by gavage, from day 0 to 14 of pregnancy.

DISCUSSION

Citrus aurantium L. (Rutaceae family), popularly known in Brazil as bitter orange, is among the species most frequently used for medicinal purposes. Phytochemical analyses of the essential oil in the fresh fruit peel of C.aurantium revealed the presence of limonene, a monoterpene as the major constituent that is commonly used as a flavoring in foods and drinks (Moraes et al. 2009, Sun 2007SUN J. 2007. d-Limonene: Safety and clinical applications. Altern Med Rev 12: 259-264.), for which it is classified in the U.S. Code of Federal Regulation as safe. Similarly, chromatogram revealed the monoterpene limonene to be the major substance, with a relative abundance of 70.75% and the molecular formula C10H16. The monoterpeneβ-Pinene, whose molecular formula is also C10H16, appears to rank second in relative abundance at 13.19% (Rozza et al. 2011). In this study, the plants used were of the same origin as those determined by Moraes et al. (2009) and Rozza et al. (2011), and therefore could have similar proportions of the active substances.

Our results showed that treatment of rats with Citrus aurantium essential oil did not interfere in maternal weight gain since the dams presented a progressive weight gain during pregnancy. In contrast, many studies suggest that C. aurantium induces weight loss effects (Cherniack 2008CHERNIACK EP. 2008. Potential applications for alternative medicine to treat obesity in an aging population. Altern Med Rev 13: 34-42.). In the present study, the essential oil of the plant was used, which contains no compounds responsible for weight loss. The substances that reduce body weight are similar to ephedra, a component of the fruit, that contains alkaloids that are adrenergic agonists and is often incorporated to supplements designed to aid in weight loss (Preuss et al. 2002PREUSS HG, DIFERDINANDO D, BAGCHI M AND BAGCHI D. 2002. Citrus aurantium as a thermogenic, weight-reduction replacement for ephedra: an overview. J Med 33: 247-264.). In some reports, Citrus aurantium products were combined with other agents, making it unclear which caused the benefit (Haaz et al. 2006).

In general, maternal deaths, reduction in body weight gain, and food and water intake are simple and sensitive indices of toxicity after exposure to toxic substances (Raza et al. 2002RAZA M, AL-SHABANAH OA, EL-HADIYAH TM AND AL-MAJED AA. 2002. Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. Sc Pharm 70: 135-145., Teo et al. 2002TEO S, STIRLING D, THOMAS S, HOBERMAN A, KIORPES A AND KHETANI V. 2002. A 90-day oral gavage toxicity study of d-methylphenidate and d,lmethylphenidate in Sprague-Dawley rats. Toxicology 174: 183-196.). In this study, the maternal weight gain did not differ. Furthermore, no alterations of daily food and water intake were observed, suggesting that the C. aurantium administration did not provoke maternal toxicity.

Effects of chemicals or drugs during pregnancy may manifest as abortions, anomalies or retarded development (Sullivan 1993SULLIVAN FM. 1993. Impact of the environment on reproduction from conception to parturition. Environ Health Persp 101: 13-18.). Our results showed non-significant differences in the number of corpora lutea, implantation sites, resorptions, live and dead fetuses, gravid uterus weight, fetal weight, and sex ratio after treatment of the dams with C. aurantium essential oil. However, litters from the rats treated with high dose presented impaired placental development, as verified by reduced placental weights. This finding could be related to limonene presence in the plant essential oil because this monoterpene presents antitumoral action (Chen et al. 2006CHEN J, LU M, JING Y AND DONG J. 2006. The synthesis of L-carvone and limonene derivatives with increased antiproliferative effect and activation of ERK pathway in prostate cancer cells. Bioorg Med Chem 14: 6539-6547.), suppressing cellular proliferation in the placenta, which could have led to the reduced placental weight. The placental index is used for verification of placental function in relation to maternal-fetal exchange (Kingdom and Kaufmann 1999KINGDOM JC AND KAUFMANN P. 1999. Oxygen and placental vascular development. Adv Exp Med Biol 474: 259-275.). In treated group with 500 mg/kg, the placental index was decreased, which can cause possible disturbances in the exchange of oxygen and nutrients from the mother to the fetuses contributing to intrauterine growth restriction, however this was not observed.

It is notable that the treated groups did not present signs of delayed ossification in the specific ossification sites (forelimbs, metatarsus, metacarpus, sternebra, and caudal vertebrae), and in total ossification sites. Chahoud and Paumgartten (2005)CHAHOUD I AND PAUMGARTTEN FJ. 2005. Relationships between fetal body weight of Wistar rats at term and the extent of skeletal ossification. Braz J Med Biol Res 38: 565-575. verified strong correlation between body weight and degree of ossification. In the present study, ossification sites and fetal weight did not alter, indicating that C. aurantium did not cause prenatal growth retardation and impaired fetal somatic development.

Prenatal administration of plants during pregnancy can result in fetal anomalies (Lemonica and Alvarenga 1994, Damasceno and Lemonica 1999, Mello et al. 2005). In our investigation, Citrus aurantium treatment did not alter the incidence of external, skeletal, and visceral anomalies. No external anomalies were found in control or treated groups. The most common skeletal anomalies in fetuses were supernumerary rib, unossified and abnormally shaped sternebra. In relation to visceral anomalies, the control and treated group presented high incidence of hydroureter, which is easily recognized because of the "S" form and complete transparency, as seen in this study. Moreover, hydroureter may or may not be associated with congenital hydronephrosis, it might possibly be a transitory variation in rodents (Taylor 1986TAYLOR P. 1986. Practical teratology. New York: Academic Press, 1st ed., 171 p.). According to the literature, approximately 20% of newborns of normal rats have this kind of visceral anomaly (Damasceno et al. 2002b, 2011a, b).

Thus, we concluded that essential oil from Citrus aurantium, administered orally to rats on GD 0 to 14, caused no teratogenic effect and did not alter maternal reproductive outcome in the doses tested.

ACKNOWLEDGMENTS

The authors are grateful to Ana Flávia Santarine Laureano, undergraduate student of the Institute of Biosciences of Botucatu - UNESP, for technical assistance.

REFERENCES

  • ALIVERTI V, BONANOMI L, GIAVINI E, LEONE VG AND MARIANI L. 1979. The extent of fetal ossification as an index of delayed development in teratogenic studies on the rat. Teratology 20: 237-242.
  • ARBO MD, SCHMITT GC, LIMBERGER MF, CHARÃO MF, MORO AM, RIBEIRO GL, DALLEGRAVE E, GARCIA SC, LEAL MB AND LIMBERGER RP. 2009. Subchronic toxicity of Citrus aurantium L. (Rutaceae) extract and p-synephrine in mice. Regul Toxicol Pharm 54: 114-117.
  • BACHMANN K, PARDOE D AND WHITE D. 1996. Scaling basic toxicokinetic parameters from rat to man. Environ Health Perspect 104: 400-407.
  • BENT S, PADULA A AND NEUHAUS J. 2004. Safety and efficacy of Citrus aurantium for weight loss. Am J Cardiol 94: 1359-1361.
  • BERGONZELLI GE, DONNICOLA D, PORTA N AND CORTHÉSY-THEULAZ IE. 2003. Essential oils as components of a diet-based approach to management of Helicobacter infection. Antimicrob Agents Ch 47: 3240-3246.
  • CHAHOUD I AND PAUMGARTTEN FJ. 2005. Relationships between fetal body weight of Wistar rats at term and the extent of skeletal ossification. Braz J Med Biol Res 38: 565-575.
  • CHEN J, LU M, JING Y AND DONG J. 2006. The synthesis of L-carvone and limonene derivatives with increased antiproliferative effect and activation of ERK pathway in prostate cancer cells. Bioorg Med Chem 14: 6539-6547.
  • CHERNIACK EP. 2008. Potential applications for alternative medicine to treat obesity in an aging population. Altern Med Rev 13: 34-42.
  • DALLAQUA B, SAITO FH, RODRIGUES T, CALDERON IM, RUDGE MV, VOLPATO GT AND DAMASCENO DC. 2013. Azadirachta indica treatment on the congenital malformations of fetuses from rats. J Ethnopharmacol 150: 1109-1113.
  • DAMASCENO DC, KISS AC, SINZATO YK, CAMPOS KE, RUDGE MV, CALDERON IP AND VOLPATO GT. 2011a. Maternal-fetal outcome, lipid profile and oxidative stress of diabetic rats neonatally exposed to streptozotocin. Exp Clin Endocrinol Diabetes 119: 408-413.
  • DAMASCENO DC AND LEMONICA IP. 1999. Embryotoxicity and anti-implantation effects of rosemary (Rosmarinus officinalis L.) extract in pregnant rats within preimplantation period. Braz J Toxicol 12: 47-54.
  • DAMASCENO DC, VOLPATO GT, FERRARI C, ROLDAN LB AND SOUZA MSS. 2008. Effect of indomethacin on the pregnant rat. Braz Arch Biol Technol 51: 79-85.
  • DAMASCENO DC, VOLPATO GT, PERSON OC, YOSHIDA A, CALDERON IMP AND RUDGE MVC. 2002b. Efeito do ácido acetilsalicílico na performance reprodutiva e na prole de ratas Wistar. Rev Assoc Méd Bras 48: 312-316.
  • DAMASCENO DC, VOLPATO GT, SARTORI TCF, RODRIGUES PF, PERIN EA, CALDERON IMP AND RUDGE MVC. 2002a. Effects of Annona squamosa extract on early pregnancy in rats. Phytomedicine 9: 667-672.
  • DAMASCENO DC, VOLPATO GT, SINZATO YK, LIMA PH, SOUZA MS, IESSI IL, KISS AC, TAKAKU M, RUDGE MV AND CALDERON IM. 2011b. Genotoxicity and fetal abnormality in streptozotocin-induced diabetic rats exposed to cigarette smoke prior to and during pregnancy. Exp Clin Endocrinol Diabetes 119: 549-553.
  • FUGH-BERMAN A AND MYERS A. 2004. Citrus aurantium, an ingredient of dietary supplements marketed for weight loss: current status of clinical and basic research. Exp Biol Med 229: 698-704.
  • GOUGEON R, HARRIGAN K, TREMBLAY J, HEDREI P, LAMARCHE M AND MORAIS JA. 2005. Increase in the thermic effect of food in women by adrenergic amines extracted from Citrus aurantium. Obes Res 13: 1187-1194.
  • HAAZ S, FONTAINE KR, CUTTER G, LIMDI N, PERUMEAN-CHANEY S AND ALLISON DB. 2006. Citrus aurantium and synephrine alkaloids in the treatment of overweight and obesity: an update. Obes Rev 7: 79-88.
  • HANSEN DK, GEORGE NI, WHITE GE, PELLICORE LS, ABDEL-RAHMAN A AND FABRICANT D. 2012. Physiological effects following administration of Citrus aurantium for 28 days in rats. Toxicol Appl Pharmacol 261: 236-247.
  • HANSEN DK, JULIAR BE, WHITE GE AND PELLICORE LS. 2011. Developmental toxicity of Citrus aurantiumin rats. Birth Defects Res B 92: 216-223.
  • KINGDOM JC AND KAUFMANN P. 1999. Oxygen and placental vascular development. Adv Exp Med Biol 474: 259-275.
  • LAPA AJ, SOUCCAR C, LIMA-LANDMAN MTR, GODINHO RO AND LIMA TCM. 2004. Farmacologia e toxicologia de produtos naturais. In: SIMÕES CMO, SCHENKEL EP, GOSMAN G, MELLO JCP, MENTZ LA AND PETROVICK PR (Eds), Farmacognosia: Da planta ao medicamento, Porto Alegre: UFRGS, Porto Alegre, Brasil, p. 247-262.
  • LEMONICA IP AND ALVARENGA CM. 1994. Abortive and teratogenic effect of Acanthospermum hispidum DC. and Cajanus cajan (L.) Millps. in pregnant rats. J Ethnopharmacol 43: 39-44.
  • MELLO FB, JACOBUS D, CARVALHO K AND MELLO JRB. 2005. Effects of Lantana camara (Verbenaceae) on general reproductive performance and teratology in rats. Toxicon 45: 459-466.
  • MONTEIRO MHD, GOMES-CARNEIRO MR, FELZENSZWALB I, CHAHOUD I AND PAUMGARTTEN FJR. 2001. Toxicological evaluation of a tea from leaves of Vernonia condensate. J Ethnopharmacol 74: 149-157.
  • MORAES TM, KUSHIMA H, MOLEIRO FC, SANTOS RC, ROCHA LR, MARQUES MO, VILEGAS W AND HIRUMA-LIMA CA. 2009. Effects of limonene and essential oil from Citrus aurantium on gastric mucosa: role of prostaglandins and gastric mucus secretion. Chem Biol Interact 180: 499-505.
  • MORAES TM, ROZZA AL, KUSHIMA H, PELLIZZON CH, ROCHA LR AND HIRUMA-LIMA CA. 2013. Healing actions of essential oils from Citrus aurantium and d-limonene in the gastric mucosa: the roles of VEGF, PCNA, and COX-2 in cell proliferation. J Med Food 16: 1162-1167.
  • POLO CM, MORAES TM, PELLIZZON CH, MARQUES MO, ROCHA LR AND HIRUMA-LIMA CA. 2012. Gastric ulcers in middle-aged rats: The healing effect of essential oil from Citrus aurantium L. (Rutaceae). Evid Based Complement Alternat Med 2012: 509451.
  • PREUSS HG, DIFERDINANDO D, BAGCHI M AND BAGCHI D. 2002. Citrus aurantium as a thermogenic, weight-reduction replacement for ephedra: an overview. J Med 33: 247-264.
  • RAZA M, AL-SHABANAH OA, EL-HADIYAH TM AND AL-MAJED AA. 2002. Effect of prolonged vigabatrin treatment on haematological and biochemical parameters in plasma, liver and kidney of Swiss albino mice. Sc Pharm 70: 135-145.
  • ROZZA AL, MORAES TM, KUSHIMA H, NUNES DS, HIRUMA-LIMA CA AND PELLIZZON CH. 2011. Involvement of glutathione, sulfhydryl compounds, nitric oxide, vasoactive intestinal peptide, and heat-shock protein-70 in the gastroprotective mechanism of Croton cajucara benth. (Euphorbiaceae) essential oil. J Med Food 14: 1011-1017.
  • RUDGE MVC, DAMASCENO DC, VOLPATO GT, ALMEIDA FGE, CALDERON IMP AND LEMONICA IP. 2007. Effect of Ginkgo biloba on reproductive outcome and oxidative stress biomarkers of streptozotocin-induced diabetic rats. Braz J Med Biol Res 40: 1095-1099.
  • SALEWSKI E. 1964. Farbemethode zum markroskopishen nachweis von implantatconsstellen na uterus der ratter naunyn schmuderbergs. Arch Pharmacol 247: 367.
  • STAPLES RE AND SCHNELL VL. 1964. Refinements in rapid clearing technique in the KOH-alizarin red S method for fetal bone. Stain Technol 39: 61-63.
  • SULLIVAN FM. 1993. Impact of the environment on reproduction from conception to parturition. Environ Health Persp 101: 13-18.
  • SUN J. 2007. d-Limonene: Safety and clinical applications. Altern Med Rev 12: 259-264.
  • TAYLOR P. 1986. Practical teratology. New York: Academic Press, 1st ed., 171 p.
  • TEO S, STIRLING D, THOMAS S, HOBERMAN A, KIORPES A AND KHETANI V. 2002. A 90-day oral gavage toxicity study of d-methylphenidate and d,lmethylphenidate in Sprague-Dawley rats. Toxicology 174: 183-196.
  • VOLPATO GT, CALDERON IM, SINZATO S, CAMPOS KE, RUDGE MV AND DAMASCENO DC. 2011. Effect of Morus nigra aqueous extract treatment on the maternal-fetal outcome, oxidative stress status and lipid profile of streptozotocin-induced diabetic rats. J Ethnopharmacol 138: 691-696.
  • VOLPATO GT, DAMASCENO DC, KEMPINAS WG, RUDGE MV AND CALDERON IM. 2009. Effect of exercise on the reproductive outcome and fetal development of diabetic rats. Reprod Biomed Online 19: 852-858.
  • VOLPATO GT, DAMASCENO DC, RUDGE MVC, PADOVANI CR AND CALDERON IMP. 2008. Effect of Bauhinia forficata aqueous extract on the maternal-fetal performance and oxidative stress biomarkers of streptozotocin-induced diabetic rats. J Ethnopharmacol 116: 131-137.
  • WILSON JG. 1965. Methods for administering agents and detecting malformations in experimental animal. In: WILSON JG AND WARKANY J (Eds), Teratology: Principles and Techniques. Chicago: University of Chicago Press, Chicago, USA, p. 262-277.

Publication Dates

  • Publication in this collection
    Mar 2015

History

  • Received
    14 July 2014
  • Accepted
    04 Nov 2014
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