Gutmann J N, Burtis W J, Dreyer B E, Andrade-Gordon P, Penzias A S, Polan M L, Insogna K L
Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, Connecticut 06510.
J Clin Endocrinol Metab. 1993 May;76(5):1314-8. doi: 10.1210/jcem.76.5.8496323.
The presence of mRNA transcripts and/or immunoreactivity for PTH-related protein (PTHrP) in several normal mammalian tissues suggests a possible paracrine or autocrine role for this hormone. Since immunohistochemical studies of human ovary demonstrate the presence PTHrP immunoreactivity in this tissue, we wondered if ovarian follicular fluid (OVFF) might contain PTHrP. We retrospectively analyzed 28 OVFF samples obtained at ova harvest in 21 women undergoing in vitro fertilization. Fourteen samples contained significant adenylate cyclase-stimulating activity in a PTHrP-sensitive bioassay. In a subsequent prospective analysis, 41 of 45 freshly obtained OVFF samples demonstrated significant activity. This bioactivity was completely neutralized by antisera to PTHrP, but was unaffected by antisera to PTH. Fifteen OVFF samples were also analyzed in a sensitive 2-site immunoradiometric assay for PTHrP, and all 15 demonstrated significant levels of the hormone. The PTHrP levels did not correlate with the presence of an ovum in the follicle or with follicular fluid calcium. Short term (24- to 48-h) cultures of granulosa-luteal cells established from 5 OVFF samples demonstrated constitutive secretion of PTHrP using the immunoradiometric assay. Neither progesterone nor estrogen affected basal secretion. RNase protection analysis of cellular RNA prepared from cultured granulosa-luteal cells demonstrated the presence of mRNA for PTHrP in these cells. We conclude that 1) human OVFF obtained after stimulation with FSH and LH contain high concentrations of PTHrP; and 2) the granulosa-luteal cell is capable of secreting PTHrP both in vivo and in vitro.
几种正常哺乳动物组织中存在甲状旁腺激素相关蛋白（PTHrP）的mRNA转录本和/或免疫反应性，提示该激素可能具有旁分泌或自分泌作用。由于对人类卵巢的免疫组织化学研究表明该组织中存在PTHrP免疫反应性，我们想知道卵巢卵泡液（OVFF）中是否可能含有PTHrP。我们回顾性分析了21名接受体外受精的女性在取卵时获得的28份OVFF样本。在一项对PTHrP敏感的生物测定中，14份样本含有显著的腺苷酸环化酶刺激活性。在随后的前瞻性分析中，45份新鲜获得的OVFF样本中有41份显示出显著活性。这种生物活性被抗PTHrP抗血清完全中和，但不受抗PTH抗血清的影响。还对15份OVFF样本进行了灵敏的双位点免疫放射分析检测PTHrP，所有15份样本均显示该激素水平显著。PTHrP水平与卵泡中卵子的存在或卵泡液钙含量无关。对从5份OVFF样本中分离的颗粒黄体细胞进行短期（24至48小时）培养，采用免疫放射分析显示这些细胞可组成性分泌PTHrP。孕酮和雌激素均不影响基础分泌。对培养的颗粒黄体细胞制备的细胞RNA进行核糖核酸酶保护分析，显示这些细胞中存在PTHrP的mRNA。我们得出结论：1）用促卵泡激素（FSH）和促黄体生成素（LH）刺激后获得的人OVFF含有高浓度的PTHrP；2）颗粒黄体细胞在体内和体外均能分泌PTHrP。
