Effect of reduced water intake on ovarian reserve, leptin immunoexpression and impact of leptin on the in vitro culture of sheep secondary follicles.

This study evaluated the effect of reduced water intake on survival, apoptosis and immunoexpression of leptin in sheep preantral follicles, activation of primordial follicles, serum levels of leptin, estradiol (E2) and progesterone (P4), and in vitro maturation (IVM) of oocytes antral follicles, as well evaluated the effects of leptin on in vitro culture of secondary follicles isolated these animals. Ewes (n = 32) were divided into four groups: water ad libitum (Control - 100%), 80%; 60% and 40% of ad libitum intake. Blood was collected to determine, leptin, E2 and P4, before and after experiment. After the slaughter, ovarian cortex was used to histological and immunohistochemistry analysis and oocytes IVM. Moreover, isolated secondary follicles were cultured in vitro for 12 days in control medium (α-MEM+) or α-MEM+ with 10 or 25 ng/mL leptin. The reduction of water intake caused a linear decreasing effect on the percentages of normal preantral follicles, especially of primordial (P < 0.05), increased the apoptosis (P < 0.05) and decreased leptin expression in preantral follicles. The treatment with 60% of water intake showed greater total growth rate of isolated secondary follicles cultured with 25 ng/L leptin (P < 0.05), compared to those cultured in α-MEM+ . In conclusion, reduced water intake impaired the number of normal sheep preantral follicles, especially of primordial follicles, increased apoptosis and decreased leptin expression in preantral follicles. Moreover, secondary follicles from of ewes that receive 60% water intake increased follicular growth after in vitro culture with 25 ng/mL leptin.

Protective effect of gallic acid on doxorubicin-induced ovarian toxicity in mouse.

The aims of the present study were to evaluate the protective effects of gallic acid against doxorubicin-induced ovarian toxicity in mice, and to verify the possible involvement of PI3K and mTOR signaling pathway members (PTEN, Akt, FOXO3a and rpS6) in the gallic acid protective actions. Mice were pretreated with NaCl (0.15 M, p.o.) (control and doxorubicin groups) or gallic acid (50, 100 or 200 mg/kg body weight, p.o.) once daily, for 5 days, and on the third day of treatment, after 1 h of treatment administration, the mice received saline solution (i.p.) (control group) or doxorubicin (10 mg/kg of body weight, i.p.). Next, the ovaries were harvested for histological (follicular morphology and activation), fluorescence (GSH and mitochondrial activity), and immunohistochemical (PCNA, cleaved caspase-3, TNF-α, p-PTEN, Akt, p-Akt, p-rpS6 and p-FOXO3a) analyses. The results showed that cotreatment with 50 mg/kg gallic acid plus doxorubicin preserved the percentage of normal follicles and cell proliferation, reduced the percentage of cleaved caspase-3 follicles, prevented inflammation, and increased GSH concentrations and mitochondrial activity compared to doxorubicin treatment alone. Furthermore, cotreatment 50 mg/kg gallic acid plus doxorrubicin increased expression of Akt, p-Akt, p-rpS6 and p-FOXO3a compared to the doxorubicin alone. In conclusion, 50 mg/kg gallic acid protects the mouse ovary against doxorubicin-induced damage by improving GSH concentrations and mitochondrial activity and cellular proliferation, inhibiting inflammation and apoptosis, and regulating PI3K and mTOR signaling pathway.

Investigation of antinociceptive, antipyretic, antiasthmatic, and spasmolytic activities of Brazilian Cannabis sativa L. roots in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Many studies are performed with the aerial parts of Cannabis sativa L. (Cannabaceae). However, roots remain poorly studied, despite citations in the scientific literature. The C. sativa roots are indicated for the treatment of pain, inflammation, fever, among other health problems. AIM OF THE STUDY: This study aimed to evaluate the antinociceptive, antipyretic, antiasthmatic, and spasmolytic activities of C. sativa roots in experimental models using mice and rats. MATERIAL AND METHODS: The chemical composition of the aqueous extract of C. sativa roots (AECsR) was evaluated by LC-MS. The antinociceptive activity was assessed in mice by the induction of writhing with acetic acid, paw licking with formalin, and reactivity in the hot plate test. Fever was induced by the administration of a suspension of Saccharomyces cerevisiae in young rats. The asthmatic activity was performed with ovalbumin (OVA)-immunized mice with cellular and histological analysis. Finally, the spasmolytic activity was performed using mice isolated trachea. For in vivo studies, the doses were 12.5, 25, or 50 mg/kg whereas for in vitro, the concentration of AECsR was 729 µg/mL. RESULTS: From the LC-MS data, we identified p-coumaroyltyramine, feruloyltyramine canabissativine in AECsR. The extract promoted a reduction of writhing in all tested doses (12.5, 25, or 50 mg/kg). Similarly, it reduced the pain in the formalin test at doses of 12.5 and 50 mg/kg (first phase) and 12.5 and 25 mg/kg (second phase). In the hot plate test, the doses of 12.5, 25, and 50 mg/kg promoted antinociceptive effect at different times, and the lowest dose maintained its action in the analyzes performed at 60, 90, and 120 min after administration. The anti-inflammatory activity of AECsR was observed in the mouse model of asthma, reducing the total leukocyte count in the bronchoalveolar fluid (BALF) at a dose of 25 mg/kg, as well as reducing eosinophilia in all tested doses (12.5, 25, and 50 mg/kg). Histological analysis of lungs stained with H&E and PAS showed a reduction in the number of inflammatory cells in the perivascular and peribronchial region, as well as reduced mucus production. CONCLUSION: The results suggest that AECsR promotes pain control, either by a central or inflammatory mechanism, and has antiasthmatic activity. However, there was no antipyretic or spasmolytic effect.

Productivity and morphological characteristics of buffel grass intercropped with butterfly pea under shade conditions / Características morfológicas e produtivas do capim buffel em consórcio com cunhã sob sombreamento

Forage production in the Brazilian semiarid region can be enhanced by irrigation and shading to maximize water use. We evaluated the productivity and morphological characteristics of buffel grass (Cenchrus ciliaris) intercropped with butterfly pea (Clitoria ternatea) under shade conditions using an irrigated area with an artificial shading system (26, 39, 55%, or full sunlight). Butterfly pea was implemented in half of the land parcels in a randomized block using a split plot design with three replications. One hundred twenty days after planting was designated as the uniformity cut and the cutting was performed every 45 days for four consecutive cycles. The morphological analysis was performed during the first and fourth cycle, and the herbage mass was determined. The shadow had a quadratic effect on the leaf appearance rate in the first cycle, while shading had a quadratic deviation effect on the leaf elongation rate in the second cycle and an increased linear effect on the length of the sheet in the first cycle. Shading had a quadratic effect on the herbage mass in the third cycle and decreased linearity in the fourth cycle. The highest percentage of buffel grass was found in the single system in the first and second cycles. In addition, we observed a decreased linear effect of shade in the first cycle and the quadratic deviation effect of shade in the second cycle. A quadratic effect of shade was observed in the third and fourth cycles. The survival of the butterfly pea had a quadratic effect with a minimum rate level of 21% shading. The crude protein content showed a system effect in four cycles with higher crude protein in the intercropping system in the first and second cycles. There was also a quadratic effect of shade in the third cycle and linear increase in the fourth cycle. In conclusion, shading increased the leaf appearance rates and elongation at intermediate levels. In addition, a high level of shading increased the leaf length and survival of butterfly pea while it decreased the mass of the forage produced.

A produção de forragem na região semiárida brasileira pode ser intensificada com a irrigação e o sombreamento, maximizando o uso da água. Com o objetivo de avaliar as características produtivas e morfológicas do Capim Buffel (Cenchrus ciliaris) consorciado com Cunhã (Clitoria ternatea), sob sombreamento, foi utilizada uma área irrigada com sistema de sombreamento artificial (26, 39, 55% ou pleno sol). Foi implantada Cunhã em metade das parcelas em delineamento de blocos casualizados em esquemas de parcelas subdivididas com três repetições. Após 120 dias foi realizado o corte de uniformização e a cada 45 dias foram realizados os cortes por quatro ciclos consecutivos, sendo realizadas análises morfológicas e determinada a massa de forragem no 1.º e 4º ciclos. Houve efeito quadrático para sombra na taxa de aparecimento foliar no 1º ciclo, efeito de desvio da quadrática para sombreamento na taxa de alongamento foliar no 2º ciclo e efeito linear crescente para sombreamento no comprimento da folha no 1º ciclo. A massa de forragem apresentou efeito quadrático para sombreamento no 3º ciclo e efeito linear decrescente no 4º ciclo. O maior percentual de Capim Buffel foi encontrado no sistema solteiro no 1º e 2º ciclos. Foi observado efeito linear decrescente para sombra no 1º ciclo e de desvio da quadrática para sombra no 2º ciclo. Já no 3º e 4º ciclos houve efeito quadrático para sombra. A sobrevivência da Cunhã apresentou efeito quadrático com taxa mínima em 21% de sombreamento. O teor de proteína bruta apresentou efeito de sistema nos quatro ciclos, sendo encontrados maiores teores de proteína bruta no sistema consorciado no 1º e 2º ciclos. Houve ainda efeito quadrático para sombra no 3º ciclo e efeito linear crescente no 4º ciclo. Conclui-se que o sombreamento promove aumento nas taxas de aparecimento e alongamento foliar em níveis intermediários. Além disso, alto nível o sombreamento promove aumento do comprimento foliar, eleva a sobrevivência da Cunhã e diminui a produção de massa de forragem.