Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing 100034, China.
Department of Obstetrics and Gynecology, Beijing Tsinghua Changgeng Hospital, Beijing 102218, China.
Chin Med J (Engl). 2017 Dec 20;130(24):2951-2959. doi: 10.4103/0366-6999.220307.
Both Mitofusin 2 (Mfn2) and pelvic organ prolapse (POP) are related to aging. The aim of the present study was to investigate the variations of Mfn2 expression in the uterosacral ligaments of patients with and/or without POP and their correlations with the expression of procollagen.
Fibroblasts were cultured using tissue specimens that were harvested from the uterosacral ligaments of POP and non-POP (NPOP) patients (n = 10 for each group) from September 2016 to December 2016. The Cell Counting Kit-8 (CCK-8) assay was used to compare the differences in cell proliferation between the two groups. Relative quantitative reverse transcription-polymerase chain reaction and Western blotting assays were employed to assess the differences in the mRNA and protein expression levels of Mfn2 and procollagen 1A1/1A2/3A1 between the two groups. The changes in procollagen expression were assessed following the downregulation of Mfn2 in the POP group using RNAi. The data were assessed with independent sample t- test or general linear model univariate analysis using the SPSS 13.0 software.
The results from CCK-8 assay indicated that cell viability in the POP group was significantly lower compared with that of the NPOP group (td5, 7, 9, 11= -5.925, -6.851, -9.129, and -9.661, respectively, all P < 0.001, from D5 to D11). The mRNA and protein expression levels of Mfn2 in the cultured fibroblasts of the POP group were significantly higher compared with those of the NPOP group (mRNA: t = 2.425, P = 0.032; protein: t = 2.392, P = 0.037, respectively), whereas only the expression levels of procollagen 1A1/1A2/3A1 were significantly higher in the NPOP group (mRNA: t = -2.165, P1A1 = 0.041; t = -2.741, P1A2 = 0.026; t = -2.147, P3A1 = 0.045, respectively; protein: t = -2.418, P1A1 = 0.029; t = -2.405, P1A2 = 0.033; t = -2.470, P3A1 = 0.012, respectively). The expression levels of procollagen in the POP group increased following the downregulation of Mfn2.
The proliferation rate and cell viability of the fibroblasts in the POP group were significantly lower compared with those in the NPOP group. In the POP fibroblasts, Mfn2 expression was increased, while procollagen expression was decreased.
线粒体融合蛋白 2（Mfn2）和盆腔器官脱垂（POP）均与衰老有关。本研究旨在探讨 Mfn2 在 POP 患者和非 POP（NPOP）患者子宫骶韧带中的表达变化及其与前胶原表达的相关性。
本研究于 2016 年 9 月至 12 月期间，采用组织样本培养法，从 POP 和 NPOP 患者（每组 10 例）的子宫骶韧带中分离出成纤维细胞。采用细胞计数试剂盒-8（CCK-8）法比较两组细胞增殖的差异。采用相对定量逆转录聚合酶链反应和 Western blot 检测法比较两组 Mfn2 和前胶原 1A1/1A2/3A1 mRNA 和蛋白表达水平的差异。采用 RNAi 下调 POP 组 Mfn2 的表达，观察前胶原表达的变化。采用 SPSS 13.0 软件进行独立样本 t 检验或单因素方差分析。
CCK-8 检测结果显示，POP 组细胞活力明显低于 NPOP 组（t d5,7,9,11= -5.925、-6.851、-9.129、-9.661，均 P < 0.001，从 D5 到 D11）。POP 组培养的成纤维细胞中 Mfn2 的 mRNA 和蛋白表达水平明显高于 NPOP 组（mRNA：t = 2.425，P = 0.032；蛋白：t = 2.392，P = 0.037），而 NPOP 组前胶原 1A1/1A2/3A1 的表达水平明显升高（mRNA：t = -2.165，P1A1 = 0.041；t = -2.741，P1A2 = 0.026；t = -2.147，P3A1 = 0.045，均 P < 0.05；蛋白：t = -2.418，P1A1 = 0.029；t = -2.405，P1A2 = 0.033；t = -2.470，P3A1 = 0.012，均 P < 0.05）。下调 Mfn2 后，POP 组前胶原的表达增加。
POP 组成纤维细胞的增殖率和细胞活力明显低于 NPOP 组。在 POP 成纤维细胞中，Mfn2 表达增加，而前胶原表达减少。
