Genotoxicity and maternal-fetal safety of the dried extract of leaves of Azadirachta indica A. Juss (Meliaceae) in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Azadirachta indica A. Juss (Meliaceae), popularly known as "neem", is used for the treatment of rheumatism, cancer, ulcers, diabetes, respiratory problems, among others. This species is present on six continents and contains more than 400 bioactive compounds. Practically all parts of the plant are used in the treatment of diseases. Although it is widely used, no study has evaluated the safety of this species throughout the gestational period in Wistar rats. AIM OF THE STUDY: To evaluate the genotoxicity and the effect of treatment with dried extract of leaves of Azadirachta indica on maternal toxicity and fetal development. MATERIALS AND METHODS: The dried extract of leaves of A. indica was obtained by spray drying after percolation of the plant material in 30% ethanol (w/w). The total flavonoids and rutin contents of the extract were determined by spectrophotometric method and HPLC-DAD, respectively. Pregnant Wistar rats (n = 40) were divided into four groups (n = 10/group): one control and three groups treated with dried extract of leaves of A. indica at doses of 300, 600 or 1200 mg/kg. Treatments were carried out from gestational day (GD) 0-20. During gestation, clinical signs of toxicity, weight gain, feed and water consumption of the dams were evaluated. On GD 21, rats were euthanized and cardiac blood was collected. Liver, kidneys, lung, heart, uterus, ovaries and bone marrow were collected. Reproductive performance parameters, histopathological analysis, biochemistry and genotoxicity were evaluated. Fetuses were evaluated for external morphology, skeletal and visceral changes. RESULTS: The total flavonoid content of the extract ranged from 2.64 to 3.01%, and the rutin content was 1.07%. There was no change in body mass gain, food and water consumption between the evaluated groups. There was also no difference between the groups in terms of biochemical parameters, reproductive performance, histopathological analysis of the mother's organs and genotoxicity. Supernumerary ossification sites of the sternum were observed, and other skeletal and visceral alterations were not significant. CONCLUSIONS: The treatment did not induce maternal toxicity, it was neither embryotoxic nor fetotoxic. The extract was not potentially genotoxic, and at a dose of 1200 mg/kg, it caused changes in the ossification of the sternum.

Evaluation of the effects of SPONDIAS MOMBIN extract as an alternative antimicrobial drug

The extract of Spondias mombin has constituents which may improve psychiatric disorders, in addition to having antiviral, antifungal, and antimicrobial activity. But despite having several benefits, it is necessary to assess whether the extract may interfere with cell metabolism so furthermore its microbicide potential can be explored. Fifteen Wistar rats were used, divided into four groups (control group; control with extract; hyperlipidemic diet; hyperlipidemic diet and extract). For 12 weeks, the animals were weighed and their blood glucose was assessed. Afterwards, they were euthanized, and the biological material was collected. The evaluation confirmed the efficacy of the extract of S. mombin against cell metabolism of rats, without negatively altering cell viability; the group of rats with an hyperlipidemic diet showed an increase in body weight; however, in the individual assessment of the organs, there were no significant changes. The glycemic index, liver parameters, lipids, and mineral ions did not show changes. Furthermore, the antimicrobial potential of S. mombin extract was observed against Staphylococcus aureus ATCC 29213 and Staphylococcus aureus BLACC. The results suggest that S. mombin extract did not interfere with cell viability, did not show cytotoxicity to cells that were exposed to it, nor did it interfere with the metabolism, organs, and biochemical indices of rats with a standard or hyperlipidemic diet. Considering such characteristics and the potential activities observed in this present study, additional evaluation should be conducted to further assess the role of S. mombin extract as a source of new alternative antimicrobial drug as well as its possible beneficial activity to the cardiovascular system.

Effects of Green Banana Biomass (Musa spp.) on Laboratory Parameters of Animal Models of Wistar Mice under Hyperlipidic Diet.

AIM: Analyze the effects of green banana biomass (Musa spp.) on the biochemical parameters in Wistar male rats under standard and hyperlipidic diet. METHODS: Ethical approval for this study was obtained from Ethics Committee on the Use of Animals CEUA - PUC-GO and consisted in the division of 32 rats in 4 differentiated groups according to their diet (standard - CT; standard with green banana biomass - CTBM; hyperlipidic - HL and hyperlipidic with green banana biomass - HLBM). Through 8 weeks animals were weighted and their glycemia were measured. After this period, they were euthanized and biological material was collected to evaluate the biochemical parameters, which analysis used the ANOVA test. RESULTS: The weekly evaluation confirmed that the efficacy of the hyperlipidic model since the final weight was bigger in the HL group in comparison with the CT group, from the sixth to the eighth week. It was also possible to observe that the CTBM group had a smaller weight compared to the CT group. Besides, the measurement of glycemia, high-density lipoprotein, triglycerides, alanine aminotransferase, aspartate aminotransferase, serum total bilirubin, direct bilirubin, alkaline phosphatase, urea, serum creatine, uric acid, calcium, phosphor, magnesium and the hematological parameters from the 8 animal of each group were compared by the ANOVA test without any significative difference (p < 0.05) in the comparisons. CONCLUSIONS: This study it did not demonstrate significative changes of the green banana biomass on the observed laboratory parameters during the 8 weeks in comparison to the standard group, indicating an absence of interference of the probiotics in the laboratory parameters on the hyperlipidic model during the analyzed period. Therefore, it is necessary an evaluation of its efficacy in obesity treatment in longer studies with molecular parameters.

Assessing the safety of using the dry extract of Justicia pectoralis Jacq. (Acanthaceae) during pregnancy of Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Justicia pectoralis Jacq. (Acanthaceae), popularly known as tilo, chambá and anador, is widely used in folk medicine in Latin American countries as a sedative, anti-anxiety, reducing menopause symptoms and in the treatment of pathologies of the respiratory tract. Although J. pectoralis is widely used by the female population, there are no studies on the safety of using this species during pregnancy. AIM OF THIS STUDY: To evaluate the effects of prenatal treatment with dry extract from the aerial parts of J. pectoralis on maternal and developmental toxicity in Wistar rats. MATERIAL AND METHODS: Pregnant Wistar rats (n = 10/group) were treated from gestational day (GD) 0-20 with the vehicle (control group) or with the dry extract of J. pectoralis at doses of 300, 600 or 1200 mg/kg. During pregnancy, clinical signs of toxicity, maternal weight, feed and water intake were evaluated. On GD 21, rats were anesthetized and intracardiac blood was collected to evaluate biochemical parameters. During cesarean section, reproductive performance parameters were recorded. The liver, kidneys, uterus and ovaries were removed for histopathological analysis. Fetuses were examined for possible malformations and/or skeletal and visceral variations. RESULTS: Treatment with dry extract of J. pectoralis did not alter weight gain, feed intake or biochemical and maternal reproductive performance parameters There were also no significant histopathological changes in the maternal organs, as well as external, skeletal and visceral malformations in the fetuses. CONCLUSION: Oral administration of J. pectoralis dry extract during pregnancy did not induce maternal toxicity or embryotoxic and teratogenic effects.

Avaliação dos parâmetros hematológicos em ratos Wistar descendentes de mães obesas / Evaluation of hematological parameters in Wistar rats descending from obese mothers

O modelo de obesidade em ratos por meio de uma dieta hiperlipídica têm se consolidado, sendo relevante compreender os componentes da hereditariedade nesta enfermidade. Por meio do hemograma é possível avaliar os elementos sanguíneos e as células responsáveis pela defesa do organismo, incluindo sua morfologia, bem como interpretando-as. Compreender esses fatores em ratos descendentes de mães obesas poderia contribuir na compreensão dos mecanismos imuno-hematológicos envolvidos na obesidade e na hereditariedade. Com isso, o objetivo deste estudo é avaliar os parâmetros hematológicos e leucocitários de ratos alimentados com dieta hiperlipídica descendentes de mães obesas. Foram utilizadas 20 ratas Wistar (peso inicial 200g) alimentadas com dieta padrão (CT) ou com dieta hiperlipídica (HL). Posteriormente, alocou-se 3 fêmeas com 1 macho por 4 dias, certificando o cruzamento. Após 23 dias do nascimento da ninhada, foi feita a separação da prole em 4 grupos com 5 animais cada, e submetidos à dieta HL ou CT. Ficando então os grupos CT, CT+ HL, HL, HL + CT. Após 20 semanas de dieta, os animais foram anestesiados e tiveram peritônio exposto para coleta de sangue, que serviu para a realização e análise do hemograma, por diferenciação celular. Todos os procedimentos foram realizados de acordo com as diretrizes da utilização de animais para fins científicos (Lei 11.794/08), e protocolada no CEUA sob o n° 9062310117. As ratas HL tiveram um ganho de peso significativamente maior a partir da 10ª semana, comparando as ratas CT. Já em relação à prole, após 13 semanas, o grupo HL obteve um ganho de peso maior em relação aos demais grupos: CT: 144,8 ± 12,15 g; CT+HL (ratos descendentes de mães CT alimentados com ração HL): 138,6 ± 5,34 g; HL+CT (ratos descendentes de mães HL alimentados com ração CT): 152,2 ± 5,23 g; HL 161 ± 10,75 g. Nos parâmetros hematológicos da prole, foi observada diferença entre o grupo CT e experimental, os monócitos apresentaram uma redução significativa de 1,6± 0,89, hemoglobina 13,8 ± 1,07, hematócrito 37,9 ± 5,51 nos grupos que receberam dieta CT comparado à CT+HL, VCM apresentou aumento de 50,4 ± 0,55 no grupo CT e 53,5 ± 0,58 na dieta HL. Dentro das condições experimentais deste trabalho e com base nos resultados obtidos, conclui-se que a dieta hiperlipídica é capaz de induzir a obesidade, com capacidade de influenciar a proliferação e a diferenciação hematopoiética, porém a prole alimentada com ração padrão (CT) consegue adaptar a dieta independente do fator hereditário em relação à obesidade.

The obesity model in rats maintained by a hyper-lipidic diet is well known, and thus, there is a need to understand the components of heredity in this disease. Using HEM, it is possible to study the blood elements and cells involved in the defense of the organism, including cell morphology and its interpretation. Understanding these factors in rats descending from obese mothers may contribute to the comprehension of the immunity and hematologic mechanisms involved in both obesity and heredity. The purpose of this study is to evaluate the hematologic and leukocyte parameters of rats fed with a hyper-lipidic diet descending from obese mothers. A total of twenty Wistar female rats (initial weight 200g) fed standard diet (SD) or hyperlipidic diet (HD) were used. Subsequently, 3 females were housed with a male for 4 days in order to get inseminated. Twenty-three days after born, the animals were separated into 4 groups, with 5 animals in each group. The groups were organized as follows: SD, SD+HD, HD, SD+HD. The animals were kept on a diet for 20 weeks. In the end of that period, animals were anesthetized and had their peritoneum exposed for the collection of blood, which was used for performing the erythrogram and leukogram analysis via cellular differentiation. All procedures were performed according the animal welfare guidelines (Law No. 11.794/08) and approved by CEUA (number 9062310117). After 10 weeks, HL mothers presented an increased body weight when compared with SD mothers. In relation to the offspring, after 13 weeks, the HD group presented greater weight gain when compared to the other groups, as follows: SD: 144.8 ± 12.15 g; SD+HD (rats from SD mothers fed with HD feed): 138.6 ± 5.34 g; HD + SD (offspring from HL mothers fed with SD feed): 152.2 ± 5.23 g; HD 161 ± 10.75 g. The offspring hematological parameters presented differences between the SD and the experimental group, where the monocytes presented a significant reduction (1.6 ± 0.89), hemoglobin (13.8 ± 1.07), hematocrit (37.9 ± 5.51) in the groups that received a SD diet compared with the SD + HD; the MCV presented an increase of 50.4 ± 0.55 in the SD group and 53.5 ± 0.58 in the group on HD diet. Conclusion: Based on the results, it can be concluded that a hyper-lipidic diet is capable of inducing obesity and may also influence hematopoietic proliferation and differentiation. However, the offspring fed with standard diet (SD) is able to adapt the diet regardless of the hereditary factor in relation to obesity.