Puerarin suppresses proliferation of endometriotic stromal cells partly via the MAPK signaling pathway induced by 17ß-estradiol-BSA.
Puerarin suppresses proliferation of endometriotic stromal cells partly via the MAPK signaling pathway induced by 17ß-estradiol-BSA.
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DOI：
10.1371/journal.pone.0045529
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Cai Z
中科院分区：
文献类型：
--
作者：
Cheng W;Chen L;Yang S;Han J;Zhai D;Ni J;Yu C;Cai Z
葛根素是从葛根中提取的一种主要的黄酮类化合物。它通过与雌激素受体（ER）结合而具有弱的雌激素作用。在早期临床治疗子宫内膜异位症的实践中，中药方剂中加入葛根，取得了较好的治疗效果。我们实验室研究了葛根素的基因组和非基因组效应。本研究旨在探讨葛根素与人胚胎干细胞（ESCs）中ER竞争性结合的能力，以及葛根素对17 β-雌二醇结合牛血清白蛋白（E2-BSA）诱导的非基因组信号通路磷酸化的影响。成功建立ESC。通过ESCs中的放射性竞争性结合试验评估葛根素与ER的结合。通过人磷酸激酶阵列筛选信号通路的激活，并通过蛋白质印迹进一步证实。根据CCK-8方案分析细胞增殖。实时荧光定量PCR和Western blotting检测细胞周期蛋白D1、考克斯-2和Cyp 19的mRNA和蛋白水平。用MEK 1/2抑制剂或ER拮抗剂证实所涉及的信号通路。我们的数据表明，葛根素与活细胞上ER的总结合能力约为17 β-雌二醇（E2）的1/3。E2-BSA能够触发ESCs中ERK 1/2的快速的、非基因组的、膜介导的激活，并且这种现象与ESCs的增殖增加相关。用葛根素处理ESCs可以消除ERK的磷酸化，显著降低细胞增殖，以及E2-BSA增强的相关基因表达水平。葛根素抑制E2-BSA诱导的ESCs增殖，部分是通过阻断一条快速的、非基因组的、膜启动的ERK通路，Cyclin D1、考克斯-2和Cyp 19的表达下调参与了这一过程。我们的数据进一步表明葛根素可能是治疗子宫内膜异位症的新候选药物。
Puerarin is a major isoflavonoid compound extracted from Radix puerariae. It has a weak estrogenic action by binding to estrogen receptors (ERs). In our early clinical practice to treat endometriosis, a better therapeutic effect was achieved if the formula of traditional Chinese medicine included Radix puerariae. The genomic and non-genomic effects of puerarin were studied in our Lab. This study aims to investigate the ability of puerarin to bind competitively to ERs in human endometriotic stromal cells (ESCs), determine whether and how puerarin may influence phosphorylation of the non-genomic signaling pathway induced by 17ß-estradiol conjugated to BSA (E2-BSA). ESCs were successfully established. Binding of puerarin to ERs was assessed by a radioactive competitive binding assay in ESCs. Activation of the signaling pathway was screened by human phospho-kinase array, and was further confirmed by western blot. Cell proliferation was analyzed according to the protocol of CCK-8. The mRNA and protein levels of cyclin D1, Cox-2 and Cyp19 were determined by real-time PCR and western blotting. Inhibitor of MEK1/2 or ER antagonist was used to confirm the involved signal pathway. Our data demonstrated that the total binding ability of puerarin to ERs on viable cells is around 1/3 that of 17ß-estradiol (E2). E2-BSA was able to trigger a rapid, non-genomic, membrane-mediated activation of ERK1/2 in ESCs and this phenomenon was associated with an increased proliferation of ESCs. Treating ESCs with puerarin abrogated the phosphorylation of ERK and significantly decreased cell proliferation, as well as related gene expression levels enhanced by E2-BSA. Puerarin suppresses proliferation of ESCs induced by E2-BSA partly via impeding a rapid, non-genomic, membrane-initiated ERK pathway, and down-regulation of Cyclin D1, Cox-2 and Cyp19 are involved in the process. Our data further show that puerarin may be a new candidate to treat endometriosis.
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影响因子：
56.9
作者：
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通讯作者：
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影响因子：
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作者：
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DOI：
10.1073/pnas.91.18.8517
发表时间：
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影响因子：
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作者：
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DOI：
10.1073/pnas.0404889101
发表时间：
2004-08-17
影响因子：
11.1
作者：
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