Suzuki T, Sasano H, Sasaki H, Fukaya T, Nagura H
Department of Pathology, Tohoku University School of Medicine, Sendai, Japan.
J Histochem Cytochem. 1994 Dec;42(12):1565-73. doi: 10.1177/42.12.7983357.
To understand changes associated with the menstrual cycle in the human ovary, it is very important to examine chronological changes in P450 aromatase (P450arom) enzymatic activity in the normal cycling ovary. Therefore, we initially examined the correlation between intensity of P450arom immunoreactivity and its biochemical enzymatic activity in five estrogen-producing human cancer cell lines (HHUA, Ishikawa, HEC-59, OMC-2, and MCF-7). P450arom immunointensity per cell was evaluated by the CAS 200 computed image analysis system, and its catalytic activity per 10(6) culture cells was analyzed by the tritiated water method. A significant correlation (r = 0.959) was demonstrated between P450arom immunoreactivity and enzymatic activity under optimal conditions of tissue fixation and immunohistochemical procedures. We then investigated P450arom immunointensity in 31 specimens of normal cycling human ovaries to examine chronological changes in P450arom activity per cell throughout the menstrual cycle. In the follicular phase, P450arom was observed in the granulosa cells of one selected antral follicle per case during the mid- to late proliferative period, and its immunointensity per granulosa cell in the follicle was not significantly different between mid- and late proliferative periods, although serum estradiol level was markedly elevated in the late proliferative period. In the luteal phase, both P450arom immunointensity per luteinized granulosa cell in a corpus luteum and serum estradiol level reached a peak in the mid-secretory period. These findings indicate that different factors may influence ovarian P450arom activity during the follicular and luteal phases, i.e., an increased number of granulosa cells in the selected follicle during the follicular phase but changes in P450arom activity per luteinized granulosa cell in the corpus luteum during the luteal phase.
为了解人类卵巢中与月经周期相关的变化，检测正常月经周期卵巢中细胞色素P450芳香化酶（P450arom）酶活性的时间变化非常重要。因此，我们首先检测了5种产生雌激素的人类癌细胞系（HHUA、Ishikawa、HEC-59、OMC-2和MCF-7）中P450arom免疫反应强度与其生化酶活性之间的相关性。通过CAS 200计算机图像分析系统评估每个细胞的P450arom免疫强度，并通过氚水法分析每10(6)个培养细胞的催化活性。在组织固定和免疫组织化学程序的最佳条件下，P450arom免疫反应性与酶活性之间显示出显著相关性（r = 0.959）。然后，我们研究了31例正常月经周期人类卵巢标本中P450arom免疫强度，以检测整个月经周期中每个细胞P450arom活性的时间变化。在卵泡期，在增殖期中期至晚期，每例中一个选定的优势卵泡的颗粒细胞中可观察到P450arom，尽管增殖期晚期血清雌二醇水平显著升高，但该卵泡中每个颗粒细胞的免疫强度在增殖期中期和晚期之间无显著差异。在黄体期，黄体中每个黄体化颗粒细胞的P450arom免疫强度和血清雌二醇水平在分泌中期均达到峰值。这些发现表明，在卵泡期和黄体期可能有不同因素影响卵巢P450arom活性，即卵泡期选定卵泡中颗粒细胞数量增加，而黄体期黄体中每个黄体化颗粒细胞的P450arom活性发生变化。
