Progesterone Increases Manganese Superoxide Dismutase Expression via a cAMP-Dependent Signaling Mediated by Noncanonical Wnt5a Pathway in Human Endometrial Stromal Cells
Progesterone Increases Manganese Superoxide Dismutase Expression via a cAMP-Dependent Signaling Mediated by Noncanonical Wnt5a Pathway in Human Endometrial Stromal Cells
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DOI：
10.1210/jc.2010-0619
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发表时间：
2010-11-01
影响因子：
5.8
通讯作者：
Sugino, Norihiro
中科院分区：
文献类型：
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作者：
Matsuoka, Aki;Kizuka, Fumie;Sugino, Norihiro
背景：锰超氧化物歧化酶 (Mn-SOD) 是线粒体中的一种抗氧化酶，通过清除人子宫内膜基质细胞 (ESC) 中的超氧自由基来保护细胞。黄体酮诱导ESCs蜕膜化过程中Mn-SOD表达增加。目的：探讨黄体酮诱导人ESCs中Mn-SOD表达的分子机制。方法：将ESCs与醋酸甲羟孕酮（MPA；10(-6)M）或二丁酰-cAMP（0.5 mM）一起孵育17 d。为了确定 cAMP 依赖性信号通路是否参与 MPA 诱导的 Mn-SOD 表达，用 H89（一种 cAMP 依赖性蛋白激酶 A 抑制剂）处理 ESC。进行染色质免疫沉淀测定以检查 cAMP 结合蛋白与 Mn-SOD 基因启动子上的 cAMP 反应元件的结合。为了检查 Wnt5a 信号传导的参与，使用抗 Wnt5a 抗体来中和 Wnt5a 活性。结果：MPA 显着增加 Mn-SOD 和 Wnt5a mRNA 水平以及细胞内 cAMP 浓度。这些增加伴随着 IGF 结合蛋白 1（蜕膜化标志物）mRNA 表达的增加。 MPA 或二丁酰-cAMP 引起的 Mn-SOD mRNA 水平的增加被 H89 完全抑制。染色质免疫沉淀分析表明，MPA 诱导 cAMP 结合蛋白与 Mn-SOD 基因启动子上的 cAMP 反应元件结合。 MPA 引起的细胞内 cAMP 浓度的增加被抗 Wnt5a 抗体处理完全抑制。 MPA 处理对 β-连环蛋白表达没有影响。结论：孕酮在蜕膜化过程中通过 cAMP 依赖性途径增加 ESC 中 Mn-SOD 的表达。黄体酮刺激的 cAMP 依赖性信号传导是由独立于 β-catenin 信号传导的非经典 Wnt5a 通路介导的。 （临床内分泌代谢杂志 95：E291-E299，2010）
Context: Manganese superoxide dismutase (Mn-SOD), an antioxidant enzyme in the mitochondria, protects cells by scavenging superoxide radicals in human endometrial stromal cells (ESCs). Mn-SOD increases in ESCs during decidualization induced by progesterone.Objective: The present study investigated the molecular mechanism for Mn-SOD expression induced by progesterone in human ESCs.Methods: ESCs were incubated with medroxyprogesterone acetate (MPA; 10(-6) M) or dibutyryl-cAMP (0.5 mM) for 17 d. To determine whether a cAMP-dependent signaling pathway is involved in the MPA-induced Mn-SOD expression, ESCs were treated with H89, an inhibitor of cAMP-dependent protein kinase A. A chromatin immunoprecipitation assay was performed to examine the binding of cAMP-binding protein to the cAMP-response element on the Mn-SOD gene promoter. To examine the involvement of Wnt5a signaling, anti-Wnt5a antibodies were used to neutralize the Wnt5a activities.Results: Mn-SOD and Wnt5a mRNA levels and intracellular cAMP concentrations were significantly increased by MPA. These increases were accompanied by an increase in the mRNA expression of IGF-binding protein-1, a marker of decidualization. The increase in Mn-SOD mRNA levels by MPA or dibutyryl-cAMP was completely inhibited by H89. The chromatin immunoprecipitation assay revealed that MPA induced cAMP-binding protein binding with cAMP-response element on the Mn-SOD gene promoter. The increase in intracellular cAMP concentrations by MPA was completely inhibited by treatment with anti-Wnt5a antibodies. MPA treatment had no effects on beta-catenin expression.Conclusions: Progesterone increased Mn-SOD expression via a cAMP-dependent pathway in ESCs during decidualization. cAMP-dependent signaling stimulated by progesterone is mediated by noncanonical Wnt5a pathways that signal independently of beta-catenin. (J Clin Endocrinol Metab 95: E291-E299, 2010)