Thaigra de Sousa Soares

Beneficial effects of Hibiscus rosa-sinensis L. flower aqueous extract in pregnant rats with diabetes

Purpose The Hibiscus rosa-sinensis flower is widely used in Brazilian traditional medicine for the treatment of diabetes and has shown antifertility activity in female Wistar rats. However, there is no scientific confirmation of its effect on diabetes and pregnancy. The aim of this study was evaluate the effect of aqueous extract of H. rosa-sinensis flowers on maternal-fetal outcome in pregnant rats with diabetes. Methods Diabetes was induced by streptozotocin (STZ, 40 mg/kg) in virgin, adult, female Wistar rats. After diabetes induction, the rats were mated. The pregnant rats were distributed into four groups (n minimum = 11 animals/group): non-diabetic, non-diabetic treated, diabetic, and diabetic treated. Oral aqueous extract of Hibiscus rosa-sinensis was administered to rats in the treatment groups during pregnancy. At term pregnancy, maternal reproductive outcomes, fetal parameters, and biochemical parameters were analyzed. Results The non-diabetic treated group showed decreased high density lipoprotein cholesterol, increased atherogenic index (AI) and coronary artery risk index (CRI), and increased preimplantation loss rate compared to the non-diabetic group. Although treatment with H. rosa-sinensis led to no toxicity, it showed deleterious effects on cardiac and reproductive functions. However, the diabetic treated group showed increased maternal and fetal weights, reduced AI and CRI, and reduced preimplantation loss rate compared to the untreated diabetic group. Conclusion Our results demonstrate beneficial effects of this flower only in pregnant rats with diabetes and their offspring. Although these findings cannot be extrapolated to human clinical use, they show that the indiscriminate intake of H. rosa-sinensis may be harmful to healthy individuals and its use should be completely avoided in pregnancy.

Effect of Himatanthus sucuuba in Maternal Reproductive Outcome and Fetal Anomaly Frequency in Rats.

The aim of this study was to evaluate the effect of Himatanthus sucuuba on the maternal reproductive outcome and fetal anomaly incidence in rats. Pregnant rats were randomly divided into three experimental groups as follows: Control = treated with water (vehicle), treated 250 = treated with H. sucuuba at dose 250 mg/kg, and treated 500 = treated with H. sucuuba at dose 500 mg/kg. The rats were orally treated, by gavage, with H. sucuuba or vehicle (water) during preimplantation and organogenic period (from gestational day 0-14). At day 21 of pregnancy, all rats were killed to obtain maternal-fetal data. The treatment with H. sucuuba at dose of 250 mg/kg caused reduction in placental efficiency and an increase preimplantation loss rate and placenta weight compared with the control. The treated 500 group presented a significant decrease in maternal weight gain, maternal weight gain minus gravid uterus weight, fetal weight, and placental efficiency compared with the control. In this group, there was a decrease in body weight at day 20 of pregnancy and metacarpus ossification and an increase in the preimplantation loss rate and skeletal anomalies compared with other groups. Himatanthus sucuuba extract caused intrauterine growth restriction, preimplantation loss, and developmental delay in the high doses tested.

O -linked N -acetyl-glucosamine deposition in placental proteins varies according to maternal glycemic levels

Aims: Hyperglycemia increases glycosylation with O‐linked N‐acetyl‐glucosamine (O‐GlcNAc) contributing to placental dysfunction and fetal growth impairment. Our aim was to determine how O‐GlcNAc levels are affected by hyperglycemia and the O‐GlcNAc distribution in different placental regions. Main methods: Female Wistar rats were divided into the following groups: severe hyperglycemia (>300 mg/dL; n = 5); mild hyperglycemia (>140 mg/dL, at least than two time points during oral glucose tolerance test; n = 7) or normoglycemia (<120 mg/dL; n = 6). At 21 days of pregnancy, placental tissue was collected and processed for morphometry and immunohistochemistry analyses, or properly stored at −80 °C for protein quantification by western blot. Key findings: Placental index was increased only in severe hyperglycemic rats. Morphometric analysis showed increased junctional zone and decreased labyrinth region in placentas exclusively from the severe hyperglycemic group. Proteins targeted by O‐GlcNAc were detected in all regions, with increased O‐GlcNAc levels in the hyperglycemic group compared to control and mild hyperglycemic rats. Proteins in endothelial and trophoblast cells were the main target for O‐GlcNAc. Whereas no changes in O‐GlcNAc transferase (OGT) expression were detected, O‐GlcNAcase (OGA) expression was reduced in placentas from the severe hyperglycemic group and augmented in placentas from the mild hyperglycemic group, compared with their respective control groups. Significance: Placental O‐GlcNAc overexpression may contribute to placental dysfunction, as indicated by the placental index. Additionally, morphometric alterations, occurring simultaneously with increased O‐GlcNAc accumulation in the placental tissue may contribute to placental dysfunction during hyperglycemia.

Safety evaluation of a vaccine: Effect in maternal reproductive outcome and fetal anomaly frequency in rats using a leishmanial vaccine as a model

While the immunogenic potential of the vaccination against infectious diseases was extensively shown, data on the safety assessment of recombinant proteins in vaccine formulations administered during pregnancy are still scarce. In the current study, the antigenicity of a vaccine against leishmaniasis (based on Leishmania braziliensis recombinant protein peroxidoxin) during pregnancy and possible maternal reproductive outcomes and fetal anomalies after immunization with a leishmanial vaccine or adjuvant alone (Bordetella pertussis derived MPLA adjuvant) were assessed. Rats were mated and allocated in three groups: Control—rats received saline; Adjuvant—rats received the adjuvant MPLA, and Vaccine—rats received the combination of MPLA and peroxidoxin. The administration was subcutaneously at the dorsal region, three times (days 0, 7, 14 of pregnancy). On day 21 of pregnancy, all rats were bled for biochemical and immunological measurements. The gravid uterus was weighed with its contents, and the fetuses were analyzed. The immunization with peroxidoxin induced a significant production of circulating IgG levels compared to other groups but caused a significant in post-implantation loss (14.7%) when compared to Control (5.0%) and Adjuvant (4.4%) groups. Furthermore, a significantly high rate of fetal visceral anomalies, such as hydronephrosis and convoluted ureter, was also observed in animals that received vaccine when compared to Control or Adjuvant groups. These data indicate the importance of safety evaluation of vaccines during pregnancy and the limited use of peroxidoxin administration during pregnancy. More importantly, the safety monitoring of immunization with MPLA derived from Bordetella pertussis demonstrated no reproductive outcomes associated with adjuvant administration, suggesting its safe use during pregnancy.

Effect of the induction of transgenerational obesity on maternal-fetal parameters

ABSTRACT Maternal obesity can cause complications for both women and their offspring for generations. Therefore, we intended to verify the repercussions of induction of transgenerational obesity on biochemical parameters, reproductive performance, and congenital anomaly frequency in Wistar rats. Female rats were used from successive generations. The female rats of parental generation (F0, n=10) were mated to obtain their offspring (F1 generation). F1 female rats received a monosodium glutamate (MSG) solution to induce obesity (n=07) or vehicle (control, n=06) during the neonatal period. These adult female rats were classified as normal or obese using the Lee Index, mated, and delivered offspring (F2 generation), which were also evaluated for obesity using the Lee Index in adult life (F2MSG, n=13, born from obese dams) or non-obesity status (F2Control, n=12, born from control dams), and were mated in adulthood. During pregnancy, glycemia and an oral glucose tolerance test (OGTT) were analyzed. At term pregnancy, the females were sacrificed for serum biochemical profile, maternal reproductive outcomes, and fetal development. In F2MSG rats, body weight gain at early pregnancy, glycemia by OGTT, total cholesterol, high-density-lipoprotein, and alanine transaminase activity were higher compared with those of F2Control rats. F2MSG rats also presented a lower implantation number and gravid uterus weight, increased pre-implantation loss and anomaly frequency in their fetuses (F3 generation) compared with those of F2Control rats. Therefore, even without significant changes in body weight gain, obesity was established at the end of pregnancy of Wistar rats using other biomarkers. Additionally, these rats showed multiple adverse reproductive outcomes, confirming the deleterious effects that lead to obesity.

Adverse effects of Croton urucurana B. exposure during rat pregnancy

ETHNOPHARMACOLOGICAL RELEVANCE Croton urucurana presents several beneficial pharmacological properties. In Brazil, women who intend to interrupt the pregnancy indiscriminately use extracts of this plant as an abortifacient agent. AIM OF STUDY To evaluated the effect of aqueous extract of Croton urucurana latex on the maternal-fetal repercussions in rats. METHODS Pregnant rats were randomly distributed into four experimental groups: Control=treated with water (vehicle); Treated 200=treated with a dose 200mg/kg; Treated 400=dose 400mg/kg and; and Treated 800=dose 800mg/kg. The rats were orally treated by gavage with Croton urucurana or vehicle (water) during whole pregnancy. At term of pregnancy, all rats were killed to obtain maternal blood and tissues samples and fetal weight and anomaly analyses. RESULTS C. urucurana treatment (Treated 400 and Treated 800) showed elevated liver enzymatic activities, reduced fetal body weight and placental efficiency. The Treated 800 group presented increased maternal total protein and cholesterol levels, and heart relative weight. All treated groups presented reduced maternal body weight and food intake, and increased pre-implantation loss rate compared to those of Control group. In addition, the treatment contributed to increased skeletal and visceral anomalies with higher doses. CONCLUSION Croton urucurana treatment caused maternal toxicity, which contributed for impairment embryo fetal development. These results showed that the indiscriminate use of plants during pregnancy should be avoided to prevent potential risk on maternal health as well as their offspring.

Effect of exercise in pregnant rats with mild diabetes on the immunological system and biochemical profiles

Materials and methods The experimental severe diabetes was induced in newborn female Wistar rats in the first day of birth by intravenous injection of Streptozotocin in a dose of 100 mg/Kg. In adult life (110 days) the rats were submitted to oral glucose tolerance test (OGTT) to confirm the moderate diabetes. After its confirmation, rats were mated and randomly assigned to 4 experimental groups (minimum n=13 animals/group): Control: non-diabetic pregnant rats without exercise (sedentary); Exercise: non-diabetic pregnant rats exposed to exercise; Diabetic: diabetic pregnant rats without exercise; Diabetic Exercise: diabetic pregnant rats exposed to exercise. The moderate intensity exercise program was swimming, from the 7th to 20th days of pregnancy. On days 0 and 17 of pregnancy, it was performed OGTT. At day 21 of pregnancy, the rats were anesthetized and the blood collected to evaluate immunological and biochemical parameters. Maternal organs (heart, liver, spleen and kidneys) were removed and immediately weighed to obtain its relative weight. Analysis of variance followed by Tukeys test were used, and the differences were considered statistically significant when p< 0.05.

Frequency of fetal anomalies from pregnant rats with mild diabetes submitted to moderate intensity exercise program

Materials and methods The experimental severe diabetes was induced in newborn female Wistar rats in the first day of birth by intravenous injection of Streptozotocin in a dose of 100 mg/Kg. In adult life (110 days) the rats were submitted to oral glucose tolerance test (OGTT) to confirm the moderate diabetes. After its confirmation, rats were mated and randomly assigned to 4 experimental groups (minimum n=13 animals/group): Control: non-diabetic pregnant rats without exercise (sedentary); Exercise: non-diabetic pregnant rats exposed to exercise; Diabetic: diabetic pregnant rats without exercise; Diabetic Exercise: diabetic pregnant rats exposed to exercise. The moderate intensity exercise program was swimming, from the 7th to 20th days of pregnancy. At day 21 of pregnancy, the rats were anesthetized and the uterus was removed. Fetuses were analyzed to external, visceral and skeletal anomalies. Analysis of variance followed by Tukey’s test was used to mean values and Fisher Exact test was used to proportions. Differences were considered statistically significant when p<0.05.

Biochemical parameters in diabetic pregnant rats treated with Bauhinia holophylla aqueous extract

Materials and methods Diabetes was induced by streptozotocin (40 mg/kg) in virgin female Wistar rats. After diabetes status confirmation, rats were mated. The pregnant diabetic rats were divided in four experimental groups (n minimum=12 animals/group): Non-diabetic; Non-diabetic Treated; Diabetic and Diabetic Treated. Oral administration of leaves aqueous extract of Bauhinia holophylla was given to non-diabetic and diabetic pregnant rats at increasing doses: 200mg/kg between day 0 and 7 of pregnancy, 400 mg/kg between day 8 and 14 and 800 mg/kg between day 15 and 21. On day 21 of pregnancy, all rats were anesthetized and killed, and the blood and liver were collected. The biochemical serum parameters (glycemic level, alanine aminotransferase [ALT], protein, cholesterol, triglycerides, High-density level lipoprotein [HDL]) and hepatic oxidative stress biomarkers (malondialdehyde [MDA], superoxide dismutase [SOD], catalase, total glutathione, thiol group) were analyze. Analysis of variance followed by Tukeys test was used. Differences were considered statistically significant when p<0.05. Results Non-diabetic and diabetic rats presented no glycemic changes. All the experimental groups showed decrease values in HDL levels compared to control group. Both diabetic groups showed higher levels of triglycerides, ALT, and MDA, and decreasing levels in catalase. Moreover, the treatment with B. holophylla in diabetic group decreased triglycerides and cholesterol levels and increased HDL compared to diabetic non-treated animals.

Effect of Hancornia speciosa aqueous extract treatment on biochemical parameters in diabetic pregnant rats

Materials and methods Diabetes was induced by streptozotocin (40 mg/Kg) in virgin female Wistar rats. After diabetes induction, rats were mated. The pregnant diabetic rats were put in four experimental groups, with n minimum=12 animals/ group: Non-diabetic; Non-diabetic Treated; Diabetic and Diabetic Treated. Hancornia speciosa leaf aqueous extract (600 mg/kg) was daily administered in all gestational period. On days 0, 7, 14 and 21 the glycemia were measured. On day 21 of pregnancy, all rats were anesthetized and killed. The blood and liver were collected. The biochemical parameters were analyzed for blood (glucose, alanine aminotransferase [ALT], total protein, total cholesterol, triglycerides, High Density Level [HDL]-cholesterol) and also in liver (malondialdehyde [MDA], superoxide dismutase [SOD], catalase, total glutathione, thiol group) for oxidative stress. Analysis of variance followed by Tukeys test were used. Differences were considered statistically significant when p<0.05. Results After treatment with H. speciosa extract, non-diabetic and diabetic rats presented no glycemic changes. Therefore, all the experimental groups showed increasing in glutathione levels compared to control group. Both diabetic groups presented higher levels of triglycerides and cholesterol and ALT actives, and also decreasing in serum protein levels compared to non-diabetic animals. Moreover, the treatment with H. speciosa in diabetic group increased HDL-cholesterol and decreased malondialdehyde (MDA) levels compared to diabetic group.