Institute of Endocrinology, Metabolism and Hypertension, Tel-Aviv Sourasky Medical Center and the Sackler Faculty of Medicine, Tel-Aviv, Israel.
J Cell Biochem. 2011 Feb;112(2):625-32. doi: 10.1002/jcb.22959.
In cultured human osteoblasts estradiol-17β (E2) modulated DNA synthesis, the specific activity of creatine kinase BB (CK), 12 and 15 lipoxygenase (LO) mRNA expression and formation of 12- and 15-hydroxyeicosatetraenoic acid (HETE). We now investigate the response of human bone cell line (SaOS2) to phytoestrogens and estrogen receptors (ER)-specific agonists and antagonists. Treatment of SaSO2 with E2, 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; ERβ-specific agonist), 4,4',4″-[4-propyl-(1H)-pyrazol-1,3,5-triyl] tris-phenol (PPT; ERα-specific agonist), biochainin A (BA), daidzein (D), genistein (G) and raloxifene (Ral) showed increased DNA synthesis and CK. Ral inhibited completely all stimulations except DPN and to some extent D. The ERα-specific antagonist methyl-piperidino-pyrazole (MPP) and the ERβ-specific antagonist 4-[2-phenyl-5,7-bis (tri-fluoro-methyl) pyrazolo [1,5-a]pyrimidin-3-yl] phenol (PTHPP) inhibited DNA synthesis, CK and reactive oxygen species (ROS) formation induced by estrogens according to their receptors affinity. The LO inhibitor baicaleine inhibited only E2, DPN and G's effects. E2 and Ral unlike all other compounds had no effect on ERα mRNA expression, while ERβ mRNA expression was stimulated by all compounds. All compounds modulated the expression of 12LO and 15LO mRNA, except E2, PPT and Ral for 12LO, and 12- and 15-HETE productions and stimulated ROS formation which was inhibited by NADPH oxidase inhibitors diphenyleneiodonium chloride (DPI) and N-acetyl cysteine and the estrogen inhibitor ICI. DPI did not affect hormonal-induced DNA and CK. In conclusion, we provide evidence for the separation of mediation via ERα and ERβ pathways in the effects of estrogenic compounds on osteoblasts, but the role of LO/HETE/ROS is unclear.
在培养的人成骨细胞中，雌二醇-17β（E2）调节 DNA 合成、肌酸激酶 BB（CK）的比活性、12 和 15 脂氧合酶（LO）mRNA 的表达以及 12-和 15-羟基二十碳四烯酸（HETE）的形成。我们现在研究植物雌激素和雌激素受体（ER）特异性激动剂和拮抗剂对人骨细胞系（SaOS2）的反应。用雌二醇、2,3-双（4-羟苯基）-丙腈（DPN；ERβ特异性激动剂）、4,4',4″-[4-丙基-（1H）-吡唑-1,3,5-三基]三苯酚（PPT；ERα特异性激动剂）、生物链素 A（BA）、大豆苷元（D）、染料木黄酮（G）和雷洛昔芬（Ral）处理 SaSO2 显示 DNA 合成和 CK 增加。Ral 完全抑制除 DPN 和在某种程度上 D 以外的所有刺激。ERα 特异性拮抗剂甲基哌啶基吡唑（MPP）和 ERβ 特异性拮抗剂 4-[2-苯基-5,7-双（三氟甲基）吡唑并[1,5-a]嘧啶-3-基]苯酚（PTHPP）根据其受体亲和力抑制雌激素诱导的 DNA 合成、CK 和活性氧（ROS）形成。LO 抑制剂黄芩素仅抑制 E2、DPN 和 G 的作用。E2 和 Ral 与所有其他化合物不同，对 ERα mRNA 表达没有影响，而所有化合物均刺激 ERβ mRNA 表达。除 E2、PPT 和 Ral 对 12LO 外，所有化合物均调节 12LO 和 15LO mRNA 的表达，并刺激 ROS 形成，而 NADPH 氧化酶抑制剂二苯基碘氯化物（DPI）和 N-乙酰半胱氨酸和雌激素抑制剂 ICI 抑制 ROS 形成。DPI 不影响激素诱导的 DNA 和 CK。总之，我们为雌激素化合物对成骨细胞的作用通过 ERα 和 ERβ 途径的分离提供了证据，但 LO/HETE/ROS 的作用尚不清楚。
