Studies on the mammary tumor-inhibiting effects of diethylstilbestrol and its mono- and diphosphate

Summary Diethylstilbestrol (DES), diethylstilbestrol monophosphate (DES-MP) and diethylstilbestrol diphosphate (DES-DP) were tested for their estrogen receptor affinity, estrogenic potency and mammary tumor-inhibiting activity in vitro and in vivo. DES had a much higher receptor binding affinity than its mono-or diphosphate. All three compounds inhibited the growth of the hormone-dependent MCF-7 and hormone-independent MDA-MB 231 breast cancer line only at relatively high concentrations. The estrogenic potency in the immature mouse uterine weight test decreased in the order DES>DES-MPDES-DP. The hormone-dependent MXT mammary tumor of the mouse was inhibited by all three compounds at a dosage of 1.0 mg/kg per week. At a dose of 0.01 mg/kg, DES, DES-MP, and DES-DP stimulated the tumor growth. Thus, for the first time, a biphasic effect on tumor growth was demonstrated in intact mature animals. As the effects of all three compounds were similar in this assay, a cleavage of the phosphate groups is likely. A decrease in estrogenic potency concomitant with a retained antitumor effect of DES-MP and DES-DP compared to DES was not demonstrable in the mature mouse using the MXT assay, only in the uterotrophic test in the immature mouse.Dedicated to Professor Dietrich Schmähl on occasion of his 60th birthdaySupported by the Deutsche Forschungsgemeinschaft and by the Verband der Chemischen Industrie, Fonds der Chemischen Industrie. The authors thank Dr. Weigert, Asta-Werke AG, Degussa Pharma Gruppe, Bielefeld, FRG, for the analysis of DES-MP and DP     

HPLC法测定己烯雌酚软膏含量

目的　应用高效液相色谱法测定己烯雌酚软膏中己烯雌酚的含量。方法　本品以乙醇超声提取,提取液进行高效液相色谱分析,色谱柱为ShimpackVP -ODS(15 0mm× 4 6mm,5 μm),流动相为甲醇 -水(75∶2 5),流速为 1 0ml·min-1,柱温35℃,检测波长 2 4 1nm。结果　平均加样回收率为 98 4 %,RSD为 0 86 %(n =5)。结论　该法简便、易行,灵敏度高,重现性好。     

快速同时检测大豆异黄酮保健食品中三种雌激素的HPLC法

〔目的〕研究检测大豆异黄酮类保健食品中三种雌激素 (雌二醇、雌酮、己烯雌酚 )含量的分析法。〔方法〕采用HypersilODS2C18色谱柱 (2 5 0mm× 4.6mmID ,5 μm) ,流动相甲醇 -水 (体积比 5 3 :47) ,流速 1.0mL/min ,检测波长 2 80nm等作为色谱条件 ,同时检测三种雌激素。〔结果〕该方法的线性范围在 0 .5～ 2 5 0mg/L ,相关系数r在 0 .998以上 ,相对标准偏差 (RSD)为 0 .9%～ 1.8% ,回收率在 91.3 %～ 98.3 %以上。〔结论〕该法具有快速、准确、灵敏等优点 ,适合大豆异黄酮中雌激素的检测     

Changes in mRNA expression of MMP-2 in the Müllerian duct of chicken embryo

Although asymmetric development of the ovary and the oviduct is a unique characteristic in birds, the mechanism of asymmetric development still remains unclear. Recently, degradation of extracellular matrix has been suggested as an important factor related to the regression of the Müllerian duct in mammals. The present study was conducted to examine a possible role of metalloproteinase-2 (MMP-2) in the regression of the right Müllerian duct in the developing chicken embryo. Morphological changes in the Müllerian ducts were studied on day 15 of incubation and mRNA expresseion of MMP-2 was studied on days 12, 15, and 18 of incubation. Morphological observation demonstrated the disappearance of basement membrane in the right Müllerian duct which undergoes the regression. RT-PCR analysis showed that MMP-2 mRNA expression of the right Müllerian duct increased on days 15 and 18 of incubation coincidently with the time of regression. In the right Müllerian duct, regression was prevented by diethylstilbestrol treatment on day 4 of incubation and a coincident decrease in MMP-2 expression was observed when compared to the control group. These results suggest that MMP-2 may be involved in the regression of the right Müllerian duct in the female embryos of the chicken.     

人参皂甙与己烯雌酚联用对去卵巢大鼠骨丢失作用的观察

目的　探讨人参皂甙单用或与己烯雌酚合用对去卵巢大鼠骨丢失的预防作用。方法　 4.5月龄 SD雌性大鼠 ,按体重随机分为正常对照组、去卵巢组、己烯雌酚组、人参皂甙组和人参皂甙与己烯雌酚联用组。采用双侧卵巢摘除术 ,体内双荧光标记法 ,胫骨上段硬组织包埋切片 ,全自动图像分析及松质骨形态计量学软件处理 ,观察药物对骨形态计量参数的影响。结果　去卵巢 1 0 w后骨量丢失和破骨细胞活性、骨形成率及骨转换率增加 ,子宫重量减轻 ,子宫内膜厚度变薄 ,存在显著性差异 (P<0 .0 1或 P<0 .0 5)。人参皂甙与己烯雌酚合用能使骨量增加 ,骨形成增多 ,骨吸收和骨转换率下降并有显著性差异 (P<0 .0 1或 P<0 .0 5) ,子宫重量和子宫内膜厚度均下降。结论　去卵巢大鼠骨量丢失 ,骨转换率增高 ,出现明显的骨质疏松 ;己烯雌酚 (1 0μg· kg- 1· d- 1 )能抑制骨吸收 ,也抑制骨形成。与 1 0 0 mg/ kg人参皂甙合用 ,能增加骨量 ,增加骨形成 ,并减轻己烯雌酚导致的子宫重量增加和内膜增厚的危险性 ,能完全预防去卵巢大鼠的骨丢失     

己烯雌酚对Wistar大鼠乳腺癌发生的干预作用

背景与目的:作为一类人工合成雌激素药物,二乙基己烯雌酚(diethyl-stilbestrol,DES)被广泛应用于治疗雌激素不足所致的更年期综合征及骨质疏松症等,但其对乳腺上皮细胞作用的研究有待进一步探讨。本研究通过建立与人相似的Wistar大鼠乳腺癌发生的模型,并用DES进行干预,观察DES在Wistar大鼠乳腺癌发生过程中的作用。方法:利用不同剂量的DES干预二甲基苯蒽[7,12-dimethylben(a)anthracene,DMBA]诱导Wistar大鼠乳腺癌模型,研究其对乳腺上皮细胞的作用。实验分为6个组:对照组、DES1组、DMBA组、DES1+DMBA组、DES2+DMBA组和DES3+DMBA组,其中DES1、DES2和DES3中DES剂量分别为:0.1mg·kg-1·d-1、0.2mg·kg-1·d-1和0.4mg·kg-1·d-1喂养大鼠,观察44周处死大鼠,取乳腺组织作常规病理分析以及核仁组成区相关嗜银蛋白(silver-binding nucleolar organizer regions,AgNOR)计数、PCNA染色强度指数(proliferating cell nuclear antigen staining intensity index,PCNA SII)、Bcl-2和C-erbB-2蛋白表达测定。结果:DMBA组和DES1+DMBA组分别有13只和2只Wistar大鼠发生癌变;单纯DES1组大鼠乳腺上皮细胞AgNOR计数及PCNASII水平上升,Bcl-2及C-erbB-2的表达提高,但未导致癌变;与其他组相比,DES2+DMBA组大鼠乳腺上皮细胞的AgNOR计数和PCNASII水平下降,Bcl-2及C-erbB-2的表达降低(P<0.05);DES3+DMBA组上述指标与DES1+DMBA组差异无统计学意义(P>0.05)。结论:单纯小剂量DES可导致大鼠乳腺上皮细胞增生,但不导致乳腺组织癌变;在DMBA诱发大鼠乳腺上皮细胞癌变过程中,小剂量DES可以促使乳腺导管上皮细胞增生、乃至癌变;而中等剂量DES可以明显抑制大鼠乳腺上皮细胞增生,加大DES剂量,抑制作用不增强。     

HPLC法测定氯霉素己烯雌酚乳膏中两主组分的含量

建立高效液相色谱法测定氯霉素己烯雌酚乳膏中氯霉素和己烯雌酚的含量。色谱柱为Kromasil C18,流动相为甲醇-水（70∶30）,流速为1.0mL.min-1,检测波长为241nm。氯霉素、己烯雌酚线性范围分别为249.1-996.5μg.mL-1,r=0.9999和4.96-19.84μg.mL-1,r=0.9999,平均回收率分别为99.81%（RSD=0.87%）和99.45%（RSD=1.03%）。该方法简便、精密度高、结果准确可靠。