Liuwei Dihuang prevents postmenopausal atherosclerosis and endothelial cell apoptosis via inhibiting DNMT1-medicated ERα methylation
Liuwei Dihuang prevents postmenopausal atherosclerosis and endothelial cell apoptosis via inhibiting DNMT1-medicated ERα methylation
复制标题
六味地黄通过抑制 DNMT1 介导的 ERα 甲基化来预防绝经后动脉粥样硬化和内皮细胞凋亡。
DOI：
10.1016/j.jep.2019.112531
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发表时间：
2020-04-24
影响因子：
5.4
通讯作者：
Bian, Huimin
中科院分区：
文献类型：
--
作者：
Chen, Qi;Zhang, Yuhan;Bian, Huimin
民族药理学相关性：经典的中药方剂六味地黄丸（LWDH）常被用于治疗更年期综合征。据报道，六味地黄丸可提高雌激素受体α（ERα）的表达以预防动脉粥样硬化（AS），然而六味地黄丸调节ERα表达的机制仍不清楚。
研究目的：揭示六味地黄丸调节ERα表达的机制。
材料与方法：检测了六味地黄丸对同型半胱氨酸（Hcy）诱导的人脐静脉内皮细胞（HUVECs）凋亡的保护作用。通过蛋白质印迹法和实时聚合酶链反应（RT - PCR）检测ERα和DNA甲基转移酶1（DNMT1）的表达。采用亚硫酸氢盐测序PCR（BSP）测定ERα基因的甲基化率。应用高效液相色谱 - 串联质谱法（HPLC - MS）测定S - 腺苷甲硫氨酸（SAM）和S - 腺苷同型半胱氨酸（SAH）的水平。在体内，将载脂蛋白E（-/-）小鼠切除卵巢以建立绝经后动脉粥样硬化（AS）模型。
结果：体外研究表明，六味地黄丸可保护人脐静脉内皮细胞免受同型半胱氨酸诱导的凋亡。六味地黄丸治疗显著提高了ERα的表达，并通过抑制DNMT1的表达降低了ERα基因的甲基化率。六味地黄丸降低了主要甲基供体SAM的水平以及SAM/SAH的比率。在体内，六味地黄丸阻止了斑块的形成并降低了同型半胱氨酸的浓度。此外，六味地黄丸在动脉粥样硬化小鼠中上调了ERα的表达，同时抑制了DNMT1的表达。
结论：六味地黄丸对绝经后动脉粥样硬化小鼠以及同型半胱氨酸处理的人脐静脉内皮细胞具有保护作用。六味地黄丸通过抑制依赖DNMT1的ERα甲基化来提高ERα的表达。
Ethnopharmacological relevance: The classical and traditional Chinese medicine prescription, Liuwei Dihuang (LWDH), has been commonly used to treat the menopausal syndrome. It has been reported that LWDH could improve estrogen receptor alpha (ER alpha) expression to prevent atherosclerosis (AS), while the mechanism of LWDH on regulating ER alpha expression was still unknown.Aim of the study: To reveal the mechanism of LWDH on regulating the ER alpha expression.Materials and methods: The protective effect of LWDH on Hcy-induced apoptosis of human umbilical vein endothelial cells (HUVECs) was examined. The expression of ER alpha and DNA methyltransferases 1 (DNMT1) were detected by Western blot and real-time polymerase chain reaction (RT-PCR). The methylation rate of the ER alpha gene was assayed by the bisulfite sequencing PCR (BSP). High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS) was applied to determine the level of S-Adenosyl methionine (SAM) and S-Adenosyl homocysteine (SAH). In vivo, the ApoE(-/-) mice were ovariectomized to establish postmenopausal atherosclerosis (AS) model.Results: In vitro study showed that LWDH protects HUVECs from Hcy-induced apoptosis. Treatment with LWDH significantly increased the ER alpha expression and reduced the methylation rate of the ER alpha gene by inhibiting the DNMT1 expression. The level of main methyl donor SAM and the ration of SAM/SAH were reduced by LWDH. In vivo, LWDH prevented the formation of plaque and reduced the concentration of Hcy. In addition, LWDH up regulated the ER alpha expression, as well as inhibiting the expression of DNMT1 in atherosclerotic mice.Conclusions: LWDH exerted protective effects on postmenopausal AS mice, and HUVECs treated with Hcy. LWDH increased of ER alpha expression via inhibiting DNMT1-dependent ER alpha methylation.