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[目的]利用小鼠胚胎成骨细胞前体细胞MC3T3-E1为细胞模型,研究雌激素与IL-6和TGF-β三者在成骨细胞分化中的具体作用机制,进一步在细胞水平探讨雌激素受体ERβ在雌激素功能中的作用。[方法]17β-雌二醇处理MC3T3-E1细胞后检测碱性磷酸酶活性、骨桥蛋白、骨保护素、骨钙素等成骨细胞相关因子的表达。同时17β-雌二醇处理后检测细胞因子IL-6和TGF-β的表达是否改变。然后分别用IL-6和TGF-β单独处理后检测MC3T3-E1细胞成骨分化相关因子和破骨细胞分化因子的表达。进一步利用siRNA干扰ERβ的表达,检测在17β-雌二醇诱导下IL-6和TGF-β表达的改变。[结果]雌激素能够诱导MC3T3-E1细胞碱性磷酸酶活性、骨桥蛋白、骨保护素、骨钙素的生成。雌激素处理后细胞因子IL-6的生成受到抑制,而TGF-β的表达上调。干扰了ERβ后,17β-雌二醇处理MC3T3-E1细胞后白细胞介素6及TGF-βmRNA的表达无明显改变。[结论]雌激素通过ERβ,调控IL-6和TGF-β的表达,进而调节成骨细胞的功能和骨的形成。
[Objective] To investigate the specific mechanism of estrogen, IL-6 and TGF-β in osteoblast differentiation by using the mouse embryonic osteoblast precursor cell MC3T3-E1 as a cell model and to further explore the mechanism of estrogen at the cellular level Role of the hormone receptor ERβ in estrogen function. [Method] The expression of alkaline phosphatase activity, osteopontin, osteoprotegerin, osteocalcin and other related factors of osteoblasts in MC3T3-E1 cells were detected by 17β-estradiol treatment. At the same time, 17β-estradiol treatment was performed to detect whether the expression of cytokines IL-6 and TGF-β changed. The expressions of osteogenic differentiation-related factors and osteoclast differentiation factors of MC3T3-E1 cells were detected by IL-6 and TGF-β alone. SiRNA was further used to interfere with the expression of ERβ, and to detect the changes of the expression of IL-6 and TGF-β under the induction of 17β-estradiol. [Results] Estrogen could induce alkaline phosphatase activity, osteopontin, osteoprotegerin and osteocalcin production in MC3T3-E1 cells. After estrogen treatment, the production of cytokine IL-6 was inhibited, while the expression of TGF-β was up-regulated. Interference with ERβ, 17β-estradiol treatment of MC3T3-E1 cells after the expression of interleukin 6 and TGF-βmRNA no significant change. [Conclusion] Estrogen regulates the function of osteoblasts and the formation of bone by regulating the expression of IL-6 and TGF-β through ERβ.