己烯雌酚对小鼠睾丸引带细胞雌激素受体的影响

目的研究雌激素受体ERα和ERβ在培养的小鼠睾丸引带细胞的表达情况,并探讨己烯雌酚(diethylstilbestrol,DES)对其表达水平的影响。方法将培养的传代小鼠睾丸引带细胞随机分为空白对照组、溶剂对照组[1μl/ml二甲基亚砜(DMSO组)]和4个实验组(终浓度分别为0.01、0.1、1、10μg/ml DES)。分别采用免疫细胞化学及Western blot方法检测雌激素受体ERα和ERβ的表达。结果免疫细胞化学显示ERα仅表达于睾丸引带细胞的细胞核,ERβ表达于细胞核及细胞质。免疫印迹显示ERα及ERβ均存在于小鼠睾丸引带细胞,且不同程度地受到DES作用剂量及作用时间的影响,以ERα表达较为明显。随着作用时间的延长,在相对低剂量组(0.01、0.1μg/ml DES组)中,ERα呈倒"U"形表达,ERβ呈"U"形表达;在相对高剂量组(10μg/ml DES组)中,除ERα高表达于12 h外,其他时间段ERα和ERβ的表达均受到明显抑制。结论 ER存在于睾丸引带细胞,不同剂量DES对睾丸引带的影响较为复杂。DES可能通过类似内源性雌激素的形式与ER结合,由ER介导引起睾丸引带组织结构和功能改变,从而影响雄性生殖系统发育。

Effects of diethylstilbestrol on estrogen receptors of gubernaculum testis cells in mice

Objective To understand the expression of estrogen receptor (ERα and ERβ) in gubernaculum testis cells in mice and the effects of diethylstilbestrol (DES) on the expression of ER.Methods The cuhured mice gubernaculum testis cells were randomly divided into six groups:normal group(without any drug),control (with 1 μl/ml DMSO) and experimental groups (treated with DES at the doses of 0.01,0.10,1.00,and 10.0 μg/ml respectively).The expression of ERα and ER3 were determined by immunocytochemistry in different groups.After 12,24 and 48 hours of treatment with DES,the expression of ER were detected by Western blot.Results Immunocytochemistry showed ERα was expressed in the nucleus of gubernaculum testis cells,but ERβ was expressed in both of the nucleus and cytoplasm.ERα and ER3 were expressed in normal mouse testes cells and influenced by different doses of DES and treatment time,especially in the expresses of ERα.At a relative lower dose group (0.01 and 0.1 μg/ml DES),ERo showed "inverted U-shaped" expression,while ERβ showed "U shaped" with time prolongation.At a relatively higher dose group (10 μg/ml DES),ERα and ERβ expressions were significantly down-regulated at all time periods except up-regulated expression of ERα at 12-hour period.Conclusion The ER expresses in the gubernaculum cell and the effects of DES with different doses on gubernaculum is complex,probably,DES may affect the development of the male reproductive system by influencing the gubernaculum tissue structure and function though a way which partly be similar to endogenous estrogens combined with ER.