Fertility and Sterility

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Fertility and Sterility Gonadotropin-releasing hormone agonist administration reduced vascular endothelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats Yoshimitsu Kitajima, M.D., Toshiaki Endo, M.D., Kengo Manase, M.D., Akira Nishikawa, M.D., Masabumi Shibuya, M.D., Ryuichi Kudo, M.D. Fertility and Sterility Volume 81, Pages 842-849 (March 2004) DOI: 10.1016/j.fertnstert.2003.11.012

FIGURE 1 Ovarian weight of control rats, hyperstimulated rats, and short-term and long-term GnRH-a treated hyperstimulated rats. Group I (control rats) ovulated by injection of 10 IU PMSG, followed by injection of 10 IU hCG 48 hours later. Group II (hyperstimulated rats) was treated with PMSG (50 IU/day) for 4 consecutive days, followed with 25 IU hCG. Group III (short-term GnRH-a treated hyperstimulated rats) received injection of GnRH-a (Leuprolide 100 μg/kg/day in saline) for 2 days beginning on the day of hCG injection. Group IV (long-term GnRH-a treated hyperstimulated rats) received injections of GnRH-a for 6 days beginning on the day of PMSG injection. Values are the mean ± SEM (n = 7 per group). *P <.05. Kitajima. Effect of GnRH-a on hyperstimulated rats.Fertil Steril 2004. Fertility and Sterility 2004 81, 842-849DOI: (10.1016/j.fertnstert.2003.11.012)

FIGURE 2 Evans blue (EB) dye content in ovaries of control rats, hyperstimulated rats, and short-term and long-term GnRH-a treated hyperstimulated rats. The ovarian Evans blue content was expressed as ng/mg tissue wet weight. Values are the mean ± SEM (n = 7 per group). *P<.05. Kitajima. Effect of GnRH-a on hyperstimulated rats.Fertil Steril 2004. Fertility and Sterility 2004 81, 842-849DOI: (10.1016/j.fertnstert.2003.11.012)

FIGURE 3 The extent of autofluorescence of Evans blue dye. (A) Ovarian surface of control rat (group I) follicles (f). (B, C) Corpus luteum (CL) and blood vessels (v) of hyperstimulated rats (group II). (D) Corpus luteum of short-term GnRH-a treated hyperstimulated rats (group III). (E, F) Follicles and blood vessels of long-term GnRH-a treated hyperstimulated rats (group IV). Bar = 100μm. Kitajima. Effect of GnRH-a on hyperstimulated rats.Fertil Steril 2004. Fertility and Sterility 2004 81, 842-849DOI: (10.1016/j.fertnstert.2003.11.012)

FIGURE 4 (A) Western blot analysis of VEGF, VEGFR-1, and VEGFR-2 protein of control rats (group I), hyperstimulated rats (group II), short-term and long-term GnRH-a treated hyperstimulated rats (groups III and group IV). Ovarian extract (100 μg of protein) was separated by commercial polyacrylamide gel and transferred to PVDF membranes using an electroblotting apparatus. (B) Protein expression was quantified by densitometric analysis using NIH Image. All densities were previously normalized against the density of the β-actin band. Values are mean ± SEM (n = 7 per group). *P<.05. Kitajima. Effect of GnRH-a on hyperstimulated rats.Fertil Steril 2004. Fertility and Sterility 2004 81, 842-849DOI: (10.1016/j.fertnstert.2003.11.012)

FIGURE 5 (A) Northern blot hybridization of VEGF, VEGFR-1, and VEGFR-2 mRNA of control rats (group I), hyperstimulated rats (group II), short-term and long-term GnRH-a treated hyperstimulated rats (group III and group IV). Total RNA was extracted from ovarian tissue of all four groups, and transferred onto nylon membranes with 32P-labeled cDNA probes. (B) Band densities are shown as bar graphs. All densities were previously normalized against the density of the L38 (ribosomal protein) band. Values are mean ± SEM (n = 7 per group). *P<.05. Kitajima. Effect of GnRH-a on hyperstimulated rats.Fertil Steril 2004. Fertility and Sterility 2004 81, 842-849DOI: (10.1016/j.fertnstert.2003.11.012)