Kozłowska Anna, Majewski Mariusz, Jana Barbara
Division of Reproductive Endocrinology and Pathophysiology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Poland.
Folia Histochem Cytobiol. 2009;47(2):257-64. doi: 10.2478/v10042-009-0043-x.
In the present study the expression pattern of the cholesterol side-chain cleavage cytochrome (P450(scc)), 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and aromatase (P450(arom)) was analyzed in the health and polycystic ovaries of gilts by means of the Western blot and immunohistochemistry. The polycystic status of ovaries was induced by i.m. dexamethasone (DXM) injections on days 7-21 of the estrous cycle. Macroscopic observation of ovaries of DXM-treated gilts revealed the presence of cysts (1-2 cm in diameter, with a mean number of 7.0+/-1.2 per ovary), a decrease (P<0.05) in number of small follicles (1-3 mm in diameter), as well as the lack of medium-sized follicles (4-6 mm in diameter) and corpora lutea, as compared to the control animals. The expression of P450(scc) (P<0.01), 3beta-HSD (P<0.05) and P450(arom) (P<0.001) proteins in the cysts was higher than in the medium-sized follicles of the control gilts. Moreover, DXM injections resulted also in an enhancement (P<0.05) in the level of P450(scc) protein in the walls of small follicles as compared to the control gilts. Following DXM administration the immunoreactivity (IR) of P450(scc) in the primordial follicles was lower than in the control group. Comparing to the control gilts, the reaction for this enzyme in DXM-treated animals was observed in secondary follicles, while for 3beta-HSD, in primordial, primary, as well as secondary follicles. The immunostaining for P450(scc) (theca cells) and P450(arom) (granulosa cells) in the small follicles of the DXM-treated gilts were more prominent than those found in the gonads of control animals. However, IR for P450(scc) was not found in the granulosa cells of small follicles in the gilts receiving DXM. The intensity of P450(scc) and P450(arom) labelling was distinctly enhanced in the cysts as compared to the medium follicles of the control animals. Furthermore, in contrary to the medium follicles of the control animals, faint IR for 3beta-HSD was found in the granulosa cell layer of cysts. Our data revealed that both the expression of P450(scc), 3beta-HSD and P450(arom) and localization of these enzymes in polycystic ovaries were different from those, found under physiological conditions. These results suggested that above-mentioned enzymes may, by influencing the ovarian steroid synthesis, play an essential role in the creation and/or course of cystic ovarian disease.
在本研究中,通过蛋白质免疫印迹法和免疫组织化学方法,分析了后备母猪健康卵巢和多囊卵巢中胆固醇侧链裂解细胞色素(P450(scc))、3β-羟基类固醇脱氢酶(3β-HSD)和芳香化酶(P450(arom))的表达模式。在发情周期的第7至21天,通过肌肉注射地塞米松(DXM)诱导卵巢出现多囊状态。对接受DXM处理的后备母猪的卵巢进行宏观观察发现,存在囊肿(直径1 - 2厘米,每个卵巢平均数量为7.0±1.2个),与对照动物相比,小卵泡(直径1 - 3毫米)数量减少(P<0.05),且缺乏中等大小卵泡(直径4 - 6毫米)和黄体。囊肿中P450(scc)(P<0.01)、3β-HSD(P<0.05)和P450(arom)(P<0.001)蛋白的表达高于对照后备母猪的中等大小卵泡。此外,与对照后备母猪相比,DXM注射还导致小卵泡壁中P450(scc)蛋白水平升高(P<0.05)。给予DXM后,原始卵泡中P450(scc)的免疫反应性(IR)低于对照组。与对照后备母猪相比,在接受DXM处理的动物的次级卵泡中观察到该酶的反应,但对于3β-HSD,在原始卵泡、初级卵泡以及次级卵泡中均有反应。在接受DXM处理的后备母猪的小卵泡中,P450(scc)(卵泡膜细胞)和P450(arom)(颗粒细胞)的免疫染色比对照动物性腺中的更明显。然而,在接受DXM的后备母猪的小卵泡颗粒细胞中未发现P450(scc)的IR。与对照动物的中等卵泡相比,囊肿中P450(scc)和P450(arom)的标记强度明显增强。此外,与对照动物的中等卵泡相反,在囊肿的颗粒细胞层中发现了微弱的3β-HSD的IR。我们的数据表明,P450(scc)、3β-HSD和P450(arom)的表达及其在多囊卵巢中的定位均与生理条件下不同。这些结果表明,上述酶可能通过影响卵巢类固醇合成,在囊性卵巢疾病的发生和/或发展过程中起重要作用。