Sex-specific response of rat costochondral cartilage growth plate chondrocytes to 17β-estradiol involves differential regulation of plasma membrane associated estrogen receptors
Sex-specific response of rat costochondral cartilage growth plate chondrocytes to 17β-estradiol involves differential regulation of plasma membrane associated estrogen receptors
复制标题
DOI：
10.1016/j.bbamcr.2012.12.022
复制
发表时间：
2013-05-01
影响因子：
5.1
通讯作者：
Schwartz, Zvi
中科院分区：
文献类型：
--
作者：
Elbaradie, Khairat B. Y.;Wang, Yun;Schwartz, Zvi
雄性和雌性大鼠生长板软骨细胞均表达雌激素受体（ER）；然而，17 β-雌二醇 (E-2) 会诱导膜反应，从而导致磷脂酶 A(2) (PLA(2))、磷脂酶 C (PLC)、前列腺素 E-2 (PGE(2)) 产生、蛋白激酶 C (PKC) 和最终丝裂原蛋白激酶 (MAPK) 的激活（仅在雌性细胞中）。这项研究调查了这些性别特异性反应是否是由于实际 ER 或下游信号传导的差异造成的。肋软骨休息区软骨细胞 (RC) 的蛋白质印迹和流式细胞术显示，雌性细胞质膜 (PM) 中的 ERot 比雄性细胞多 2-3 倍。衣霉素阻断 E2 依赖性 ER 易位至 PM，表明需要棕榈酰化。免疫共沉淀显示 E-2 仅在雌性 RC 中诱导 ER 异构体之间的复合物形成。为了检查男性 PKC 和 PGE(2) 缺乏反应是否是由于信号传导差异所致，我们检查了 ER 的参与以及 PLC 和 PLA(2) 的作用。选择性 ER α（丙基吡唑三醇，PPT）和 ER β（二芳基丙腈，DPN）激动剂仅在女性 RC 中激活 PKC。 PLC 抑制剂 U73122 阻断 E-2 对 PKC 的作用，而胞质 PIA(2) 抑制剂 AACOCF(3) 抑制女性 RC 中对 PGE(2) 的作用，证实了 PLC 和 PLA(2) 参与该机制。 PLC 激活剂、m-3M3F beta S 激活的 PKC 和 PLAA 肽增加了男性和女性 RC 中的 PGE(2) 水平，表明信号传导通路的存在。这些数据表明膜 ER 数量、定位、易位和相互作用的差异是对 E-2 的两性二态性反应的原因。 (C) 2013 Elsevier B.V. 保留所有权利。
Both male and female rat growth plate chondrocytes express estrogen receptors (ERs); however 17 beta-estradiol (E-2) induces membrane responses leading to activation of phospholipase A(2) (PLA(2)), phospholipase C (PLC), prostaglandin E-2 (PGE(2))production, protein kinase C (PKC), and ultimately mitogen protein kinase (MAPK) only in female cells. This study investigated if these sex-specific responses are due to differences in the actual ERs or in downstream signaling. Western blots and flow cytometry of costochondral cartilage resting zone chondrocytes (RCs) showed 2-3 times more ERot in plasma membranes (PMs) from female cells than male cells. Tunicamycin blocked E2-dependent ER-translocation to the PM, indicating palmitoylation was required. Co-immunoprecipitation showed E-2 induced complex formation between ER isoforms only in female RCs. To examine if the lack of response in PKC and PGE(2) in males is due to differences in signaling, we examined involvement of ERs and the role of PLC and PLA(2). Selective ER alpha (propylpyrazole triol, PPT) and ER beta (diarylproprionitrile, DPN) agonists activated PKC in female RCs only. The PLC inhibitor, U73122 blocked E-2's effect on PKC and the cytosolic PIA(2) inhibitor, AACOCF(3) inhibited the effect on PGE(2) in female RCs, confirming involvement of PLC and PLA(2) in the mechanism. The PLC activator, m-3M3F beta S activated PKC and PLAA peptide increased PGE(2) levels in male and female RCs, showing that the signaling pathways are present. These data indicate that differences in membrane ER amount, localization, translocafion and interaction are responsible for the sexual dimorphic response to E-2. (C) 2013 Elsevier B.V. All rights reserved.