Hou Ning Ning, Zhu Yi Min, Huang He Feng
Department of Obstetrics and Gynecology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou 310006.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2006 Aug;39(4):289-96.
In this present study, female rats were ovariectomized (OVX) as the models of osteoporosis. The aim is to determine the different mechanisms of estrogen receptor(ER) alpha and beta pathway in mediating estrogen to participate in trabecular bone metabolism, and to further explore the distinction of modulation on ER alpha or ER beta between estrogens with different components. Mature female Sprague-Dawley rats (n=40) were randomly divided into four groups: group Control (sham operated), group OVX (only ovariectomized), group CEE (OVX rats treated with conjugated equine estrogens) and group EV (OVX rats treated with estradiol valerate). Sham operation and OVX were performed 48 days (12 estrums) before different liquid diet. The rats in group Control and group OVX were orally administrated with physiological saline solution and the rats in group CEE or group EV were orally administrated with CEE or EV for 12 days (3 estrums) before sacrifice. Relative quantitative reverse transcription- polymerase chain reaction (RT-PCR) and western blot techniques were utilized to compare the levels of ER alpha and ER beta mRNA and proteins in trabecular bone among groups. The results showed that in rat trabecular bone of group Control, the expression of ER alpha protein (1.433 +/- 0.250) was significantly higher than that of ER beta(0.687 +/- 0.120), whereas the ER alpha mRNA (0.285 +/- 0.033) was much lower than ERbeta mRNA(0.590 +/- 0.044). Following OVX, the levels of ER alpha protein (0.685 +/- 0.103) declined significantly, whereas mRNA levels (0.405 +/- 0.036) markedly increased. Both the protein (1.091 +/- 0.078) and mRNA (0.729 +/- 0.030) levels of ER beta significantly increased after OVX. After treatment with CEE, the expression of ER beta protein (0.583 +/- 0.129) and mRNA (0.618 +/- 0.043) were markedly down-regulated compared with group OVX. After treatment with EV, the ER alpha protein expression (1.272 +/- 0.247) was markedly up-regulated, while ERa mRNA (0.277 +/- 0.040) and ER beta protein (0.620 +/- 0.174) were markedly down-regulated compared with group OVX. On the other hand, the loss of bone mass in rat trabecular bone by OVX (0.192 +/- 0.010) was markedly gained after CEE (0.228 +/- 0.012) and EV (0.259 +/- 0.007) treatment. These results demonstrate that ERalpha may be the predominant subtype expressed in female rat trebecular bone and play a key role in the bone metabolism. This study also shows that estrogens with different components protect bone through variant ERs pathway.
在本研究中,将雌性大鼠去卵巢(OVX)作为骨质疏松症模型。目的是确定雌激素受体(ER)α和β途径介导雌激素参与小梁骨代谢的不同机制,并进一步探索不同成分雌激素对ERα或ERβ调节作用的差异。将40只成熟雌性Sprague-Dawley大鼠随机分为四组:对照组(假手术)、OVX组(仅去卵巢)、CEE组(用结合马雌激素治疗的OVX大鼠)和EV组(用戊酸雌二醇治疗的OVX大鼠)。在给予不同液体饮食前48天(12个发情周期)进行假手术和OVX手术。对照组和OVX组大鼠口服生理盐水,CEE组或EV组大鼠在处死前12天(3个发情周期)口服CEE或EV。采用相对定量逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹技术比较各组小梁骨中ERα和ERβ mRNA及蛋白质水平。结果显示,在对照组大鼠小梁骨中,ERα蛋白表达(1.433±0.250)显著高于ERβ(0.687±0.120),而ERα mRNA(0.285±0.033)远低于ERβ mRNA(0.590±0.044)。OVX后,ERα蛋白水平(0.685±0.103)显著下降,而mRNA水平(0.405±0.036)显著升高。OVX后ERβ的蛋白(1.091±0.078)和mRNA(0.729±0.030)水平均显著升高。CEE治疗后,与OVX组相比,ERβ蛋白(0.583±0.129)和mRNA(0.618±0.043)表达显著下调。EV治疗后,与OVX组相比,ERα蛋白表达(1.272±0.247)显著上调,而ERα mRNA(0.277±0.040)和ERβ蛋白(0.620±0.174)显著下调。另一方面,OVX导致大鼠小梁骨骨量丢失(0.192±0.010),CEE(0.228±0.012)和EV(0.259±0.007)治疗后骨量显著增加。这些结果表明,ERα可能是雌性大鼠小梁骨中表达的主要亚型,在骨代谢中起关键作用。本研究还表明,不同成分的雌激素通过不同的ER途径保护骨骼。