Department of Pathology and Pathophysiology, Xiangya Hospital, Central South University, Changsha, China.
Key Laboratory of Tumor Immunology and Microenvironmental Regulation, Guilin Medical University, Guilin, China.
J Cell Mol Med. 2020 Jan;24(1):875-885. doi: 10.1111/jcmm.14797. Epub 2019 Nov 19.
Formononetin is a natural isoflavone compound found mainly in Chinese herbal medicines such as astragalus and red clover. It is considered to be a typical phytooestrogen. In our previous experiments, it was found that formononetin has a two-way regulatory effect on endothelial cells (ECs): low concentrations promote the proliferation of ECs and high concentrations have an inhibitory effect. To find a specific mechanism of action and provide a better clinical effect, we performed a structural transformation of formononetin and selected better medicinal properties for formononetin modifier J1 and J2 from a variety of modified constructs. The MTT assay measured the effects of drugs on human umbilical vein endothelial cell (HUVEC) activity. Scratch and transwell experiments validated the effects of the drugs on HUVEC migration and invasion. An in vivo assessment effect of the drugs on ovariectomized rats. Long-chain non-coding RNA for EWSAT1, which is abnormally highly expressed in HUVEC, was screened by gene chip, and the effect of the drug on its expression was detected by PCR after the drug was applied. The downstream factors and their pathways were analysed, and the changes in the protein levels after drug treatment were evaluated by Western blot. In conclusion, the mechanism of action of formononetin, J1 and J2 on ECs may be through EWSAT1-TRAF6 and its downstream pathways.
芒柄花素是一种天然异黄酮化合物，主要存在于中药如黄芪和红三叶草中。它被认为是一种典型的植物雌激素。在我们之前的实验中，发现芒柄花素有双向调节内皮细胞（ECs）的作用：低浓度促进 ECs 的增殖，高浓度则有抑制作用。为了找到一个特定的作用机制，并提供更好的临床效果，我们对芒柄花素进行了结构转化，从多种修饰结构中选择了更好的药用性质的芒柄花素修饰剂 J1 和 J2。MTT 检测法测量药物对人脐静脉内皮细胞（HUVEC）活性的影响。划痕和 Transwell 实验验证了药物对 HUVEC 迁移和侵袭的影响。药物对去卵巢大鼠的体内评估作用。通过基因芯片筛选出异常高表达于 HUVEC 的 EWSAT1 的长链非编码 RNA，并在应用药物后通过 PCR 检测药物对其表达的影响。分析下游因子及其通路，并通过 Western blot 评估药物处理后蛋白水平的变化。总之，芒柄花素、J1 和 J2 对 ECs 的作用机制可能是通过 EWSAT1-TRAF6 及其下游通路。
