Ere stained by May-Gr wald solution (1 w/v) followed byLienou et
Ere stained by May-Gr wald solution (1 w/v) followed byLienou et al. BMC Complementary and Alternative Medicine 2012, 12:36 http://www.biomedcentral.com/1472-6882/12/Page 3 ofGiemsa solution (1 w/v) and viewed under low magnification (40 ? under a light microscope. This staining helped in characterizing each phase of the rat’s estrous cycle, its length and that of the complete cycle. At the end of the experimental period, 6 animals in each group were randomly sacrificed by anesthesia using chloroform. Their ovaries and uteri were removed, blotted, weighed and stored at – 20 until use. The remaining rats (7 per group) were crossed the following day, during two weeks, with males of proven fertility. Chaetocin side effects vaginal smears were collected on a daily basis in order to assess for the presence of sperm. A laparoscopy was undertaken under diazepam (5 mg/ml, 5 mg/kg) and Ketamin (50 mg/ml, 80 mg/kg) ten days after the day of mating to PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/27196668 count the number of implantation sites in uterine cords and the number of corpora lutea in ovaries. After delivery, the fetuses were weighed and their number recorded. From these data, the number of resorption sites (number of implantation site – number of live fetuses), implantation index ([total number of implantation sites/number corpora lutea] ?100), resorption index ([total number of resorption sites/total number of implantation sites] ?100), preimplantation loss ([number of corpora lutea – number of implantations/ number of corpora lutea] ?100), postimplantation loss ([number of implantations ?number of life fetuses/ number of implantations] ?100), antifertility activity ([number of females without life fetuses/total number of females] ?100), antiimplantation activity ([number of females without implantation sites/total number of females] ?100), and gestation rate ([number of females with life fetuses at birth/total number of gestational females] ?100) were calculated [28].Preparation of the uterine and ovarian supernatants and biochemical analysisResultsEffect of AESb on body weight gain and food intakeThe effect of AESb on the body weight of female rats during the treatment is presented in Figure 1. There was a linear increase, at various rates, in their growth. Compared to control animals, a significant drop in the body weight of animals treated with the doses of 32 and 64 mg/kg was indeed noticed after 19 days of treatment. Animals receiving the 8 mg/kg dose instead gained more weight (p < 0.05) compared to control animals, starting from the 25th day of treatment till the end of PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/27864321 the experiment. As concerns the monitoring of their food intake, no significant variation, whatever the duration of the treatment, was observed between the different experimental groups (Figure 2).Effect of AESb on the age and estrous cycle phases at vaginal openingOvaries and uteri were homogenized in Tris – sucrose buffer (0.25 M sucrose, 1 mM EDTA and 10 mM TrisHCl, pH 7.4) at 1 and 2 respectively. The homogenate was then centrifuged at 6000 ?g at 4 (Beckman model J2-21) for 15 min, and the supernatants collected were used for protein [29] and cholesterol [30-32] assays.Statistical analysisFigure 3 shows the mean age of animals at vaginal opening and the percentage of those presenting vaginal aperture at a given age. Female rats that received AESb at the two highest doses presented vaginal opening almost two days earlier (p < 0.05) as compared to the control animals [43.33 ?0.73 days (0 mg/kg) vs 41.25 ?0.51 days (32 mg/kg) or 41.4.