Department of Gynecology and Obstetrics, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
Department of Anesthesiology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430060, P.R. China.
Int J Mol Med. 2017 Aug;40(2):347-356. doi: 10.3892/ijmm.2017.3042. Epub 2017 Jun 26.
The present study aimed to reveal the metabolic alterations of the extracellular matrix (ECM) in uterosacral ligament (USL) with pelvic organ prolapse (POP) and to explore the role of transforming growth factor‑β1 (TGF‑β1) in pathogenesis of POP. For this purpse, 60 participants who underwent hysterectomy for benign indications were enrolled, 30 of which had symptomatic POP (grade II, III or IV) and composed the POP group, and the other 30 had asymptomatic POP (grade I or less) and served as the controls. Collagen fibers, elastin,matrix metalloproteinase (MMP)‑2/9, tissue inhibitor of matrix metalloproteinases (TIMP)‑2 and TGF‑β1 were examined by Masson's trichrome staining, immunohistochemistry and RT-qPCR using USL biopsies. In vitro, human USL fibroblasts (hUSLFs) were primary cultured, pre-treated with recombinant TGF‑β1 (0, 5, or 10 ng/ml) and then subjected to cyclic mechanical stretching (CMS; 0 or 5,333 µε strain). Changes in the expression levels of collagen type I/III, elastin, TIMP‑2, MMP‑2/9 and Smad were detected. Our results revealed that at the tissue level, the expression of collagen fibers, elastin, TIMP‑2 and TGF‑β1 was significantly reduced in the POP group, while the activities of MMP‑2/9 were significantly upregulated, compared with the control group. Statistical analysis indicated that the mRNA expression of TGF‑β1 inversely correlated with the severity of POP partially. Our in vitro experimental data demonstrated that a CMS of 5333 µε strain promoted the degradation of ECM proteins, inhibited the synthesis of TIMP‑2, and upregulated the proteolytic activities of MMP‑2/9. Pre-treatment with TGF‑β1 attenuated the loss of ECM by stimulating the synthesis of TIMP‑2 and inhibiting the activities of MMP‑2/9 through the TGF‑β1/Smad3 signaling pathway. On the whole, our data indicate that the reduced anabolism and increased catabolism of ECM proteins in USL are the pathological characteristics of POP. TGF‑β1 not only has a specific value in predicting the severity of POP, but should also be considered as a novel therapeutic target for POP.
本研究旨在揭示患有盆腔器官脱垂(POP)的子宫骶韧带(USL)中细胞外基质(ECM)的代谢改变,并探讨转化生长因子-β1(TGF-β1)在 POP 发病机制中的作用。为此,纳入了 60 名因良性疾病接受子宫切除术的患者,其中 30 名患有有症状的 POP(II 级、III 级或 IV 级),组成 POP 组,另 30 名患有无症状的 POP(I 级或更轻),作为对照组。通过 Masson 三色染色、免疫组织化学和 RT-qPCR 检测 USL 活检中胶原蛋白纤维、弹性蛋白、基质金属蛋白酶-2/9(MMP-2/9)、基质金属蛋白酶抑制剂-2(TIMP-2)和 TGF-β1。体外原代培养人 USL 成纤维细胞(hUSLFs),用重组 TGF-β1(0、5 或 10ng/ml)预处理,然后进行周期性机械拉伸(CMS;0 或 5,333µε 应变)。检测胶原蛋白 I/III、弹性蛋白、TIMP-2、MMP-2/9 和 Smad 的表达水平变化。我们的结果表明,在组织水平上,与对照组相比,POP 组中胶原蛋白纤维、弹性蛋白、TIMP-2 和 TGF-β1 的表达显著降低,而 MMP-2/9 的活性显著上调。统计分析表明,TGF-β1 的 mRNA 表达与 POP 的严重程度呈负相关。我们的体外实验数据表明,5333µε 应变的 CMS 促进 ECM 蛋白降解,抑制 TIMP-2 合成,并上调 MMP-2/9 的蛋白水解活性。TGF-β1 预处理通过刺激 TIMP-2 的合成并抑制 MMP-2/9 的活性,通过 TGF-β1/Smad3 信号通路减弱 ECM 的丢失。总的来说,我们的数据表明,USL 中 ECM 蛋白的合成减少和分解增加是 POP 的病理特征。TGF-β1 不仅在预测 POP 的严重程度方面具有特定的价值,而且还应被视为 POP 的一种新的治疗靶点。