Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing, China.
Front Endocrinol (Lausanne). 2020 Nov 25;11:586242. doi: 10.3389/fendo.2020.586242. eCollection 2020.
To assess the effects of 17β-estradiol (E2) on proliferation, apoptosis, and protein expressions of fibroblasts at different concentrations and time intervals to reveal the mechanism of E2 in the treatment of pelvic organ prolapse (POP).
The uterosacral ligament fibroblasts were collected from seven POP patients for primary culture of fibroblasts. The culture media containing 0, 10, 10, 10, and 10 mol/L E2 were used for 24, 48, 72, and 96 h.
The cells were collected for cell counting kit-8 (CCK-8), apoptosis, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and Western blotting assays.
Compared with the control group, in the values of fibroblasts cultured in 10 mol/L E2 for 72 h, the proliferation, mRNA and protein expression of Mitofusin-2 (Mfn2) separately increased (P < 0.05), decreased (P<0.001) and decreased (P<0.001). However, the expression level of procollagen 1A1/1A2/3A1 and cyclinD1 markedly increased (P<0.001, all), which was consistent with the results of protein level. What's more, the expression of estrogen receptor α(ERα), estrogen receptor β(ERβ) and G protein-coupled receptor 30(GPR30) were significantly increased in 10 mol/L E2 group.
E2 can inhibit the progress of POP by inhibiting the expression level of Mfn2, as well as promoting expression of procollagens and proliferation of fibroblasts. This effect is time- and concentration-dependent. Only when the estrogen concentration reaches 10 mol/L, the therapeutic effect is the greatest after 72 h.
评估不同浓度和时间间隔的 17β-雌二醇（E2）对成纤维细胞增殖、凋亡和蛋白表达的影响，揭示 E2 治疗盆腔器官脱垂（POP）的机制。
从 7 名 POP 患者中采集子宫骶骨韧带成纤维细胞进行原代培养。使用含 0、10、10、10 和 10 mol/L E2 的培养基分别培养 24、48、72 和 96 h。
收集细胞进行细胞计数试剂盒-8（CCK-8）、细胞凋亡、实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测。
与对照组相比，在 10 mol/L E2 培养 72 h 的成纤维细胞中，Mfn2 的增殖、mRNA 和蛋白表达分别增加（P < 0.05）、减少（P<0.001）和减少（P<0.001）。然而，前胶原 1A1/1A2/3A1 和细胞周期蛋白 D1 的表达水平明显增加（P<0.001，均），这与蛋白水平的结果一致。此外，在 10 mol/L E2 组中，雌激素受体 α（ERα）、雌激素受体 β（ERβ）和 G 蛋白偶联受体 30（GPR30）的表达显著增加。
E2 通过抑制 Mfn2 的表达水平，同时促进前胶原的表达和成纤维细胞的增殖，从而抑制 POP 的进展。这种作用具有时间和浓度依赖性。只有当雌激素浓度达到 10 mol/L 时，72 h 后才达到最大治疗效果。
