Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, People's Republic of China.
Department of Obstetrics and Gynecology, Cixi Maternity and Child Health Hospital, Ningbo 315300, Zhejiang, People's Republic of China.
Drug Des Devel Ther. 2020 Sep 21;14:3815-3826. doi: 10.2147/DDDT.S265066. eCollection 2020.
To evaluate the effects of quercetin on proliferation, invasion and migration of endometrial stromal cells (ESCs) from adenomyosis patients.
Primary ectopic ESCs (EESCs) and eutopic ESCs (EuESCs) were obtained and purified from patients undergoing total hysterectomy for adenomyosis and identified by immunocytochemistry staining. The cytotoxicity and inhibition rate were determined by CCK-8 assay to obtain the IC value. Cell proliferative, migratory, and invasive abilities were detected by BrdU, wound scratch, transwell assays, respectively. Western blot analysis was employed to explore the effects of quercetin on the expression of MMP-2, MMP-9, Ezrin and Fascin proteins in cells.
Both EESCs and EuESCs were characterized with strongly positive staining for vimentin and almost negative for cytokeratin. Quercetin inhibited the viability of EESCs and EuESCs in a dose- and time-dependent manner, with an IC = 33.00 μM for EuESCs and IC = 74.88 μM for EESCs at 72 h. Thus, the final concentrations and action time of quercetin in EuESCs (0, 20, 40, and 80 μM for 72 h) and EESCs (0, 40, 80, and 160 μM for 72 h) were selected. BrdU assay showed that quercetin dose-dependently suppressed the proliferation of EESCs and EuESCs, while the inhibition rate in EESCs was higher. Similarly, administration of quercetin in EESCs and EuESCs significantly decreased the motility and invasiveness in a dose-dependent fashion, with stronger inhibitory effects on EESCs. Finally, Western blot analysis demonstrated that invasion- and migration-related proteins (MMP-2, MMP-9, Erzin, and Fascin) were significantly downregulated with the quercetin concentration increasing. Moreover, the decreased level of these proteins in EESCs under quercetin exposure was greater than that in EuESCs.
Quercetin can inhibit the proliferation of EESCs in adenomyosis and reduce their mobility and invasiveness. These inhibitory effects may be related to the downregulation of MMP-2, MMP-9, Fascin, and Erzin proteins.
评估槲皮素对子宫腺肌病患者子宫内膜基质细胞（ESCs）增殖、侵袭和迁移的影响。
从接受全子宫切除术治疗子宫腺肌病的患者中获得并纯化原代异位 ESCs（EESCs）和在位 ESCs（EuESCs），并通过免疫细胞化学染色进行鉴定。通过 CCK-8 测定确定细胞毒性和抑制率，以获得 IC 值。通过 BrdU、划痕实验和 Transwell 实验分别检测细胞的增殖、迁移和侵袭能力。Western blot 分析用于探讨槲皮素对细胞中 MMP-2、MMP-9、Ezrin 和 Fascin 蛋白表达的影响。
EESCs 和 EuESCs 均表现出波形蛋白强烈阳性染色，而细胞角蛋白几乎阴性。槲皮素呈剂量和时间依赖性地抑制 EESCs 和 EuESCs 的活力，在 72 小时时 EuESCs 的 IC = 33.00 μM，EESCs 的 IC = 74.88 μM。因此，选择了槲皮素在 EuESCs（0、20、40 和 80 μM 作用 72 小时）和 EESCs（0、40、80 和 160 μM 作用 72 小时）中的最终浓度和作用时间。BrdU 实验表明，槲皮素呈剂量依赖性地抑制 EESCs 和 EuESCs 的增殖，而 EESCs 的抑制率更高。同样，槲皮素在 EESCs 和 EuESCs 中的给药显著降低了迁移和侵袭的活力，呈剂量依赖性，对 EESCs 的抑制作用更强。最后，Western blot 分析表明，侵袭和迁移相关蛋白（MMP-2、MMP-9、Ezrin 和 Fascin）随槲皮素浓度的增加而显著下调，并且 EESCs 在槲皮素暴露下这些蛋白的下调水平大于 EuESCs。
槲皮素可抑制子宫腺肌病患者 EESCs 的增殖，并降低其迁移和侵袭能力。这些抑制作用可能与 MMP-2、MMP-9、Fascin 和 Ezrin 蛋白的下调有关。
