Klotho Protein Reduced the Expression of Matrix Metalloproteinase-1 (MMP-1) and Matrix Metalloproteinase-3 (MMP-3) in Fibroblasts from Patients with Pelvic Organ Prolapse (POP) by Down-Regulating the Phosphorylation of ERK1/2
Klotho Protein Reduced the Expression of Matrix Metalloproteinase-1 (MMP-1) and Matrix Metalloproteinase-3 (MMP-3) in Fibroblasts from Patients with Pelvic Organ Prolapse (POP) by Down-Regulating the Phosphorylation of ERK1/2
复制标题
Klotho 蛋白通过下调 ERK1/2 磷酸化降低盆腔器官脱垂 (POP) 患者成纤维细胞中基质金属蛋白酶-1 (MMP-1) 和基质金属蛋白酶-3 (MMP-3) 的表达
DOI：
10.12659/msm.913623
复制
发表时间：
2019-05-22
影响因子：
3.1
通讯作者：
Chen, Xinliang
中科院分区：
文献类型：
--
作者：
Qiu, Jun;Qin, Menglu;Chen, Xinliang
背景：盆腔器官脱垂（POP）是由于年龄相关的萎缩和盆底组织的削弱，胶原蛋白和细胞外基质（ECM）被金属蛋白酶（MMPs）降解。本研究旨在探讨由KL基因编码的年龄相关酶klotho在从POP患者获得的培养成纤维细胞中的作用以及活性氧（ROS）、白细胞介素-6 （IL-6）和MMPs的水平。材料/方法：从3例POP患者和3例正常人的结缔组织中获得盆底成纤维细胞。采用细胞计数试剂盒-8 （CCK-8）和流式细胞术检测细胞增殖和ROS生成。采用酶联免疫分析法（ELISA）、定量实时聚合酶链式反应（qRT-PCR）和Western blot检测白细胞介素-6 （IL-6）、klotho、金属蛋白酶-1 （MMP-1）、MMP-3、细胞外信号调节激酶1/2 （ERK1/2）和p-ERK1/2的水平。结果：在体外培养的POP患者盆底成纤维细胞中，与正常成纤维细胞相比，POP患者成纤维细胞中klotho蛋白和klotho mRNA的表达明显下调。在POP患者的培养成纤维细胞中补充Klotho可以促进细胞生长，减少ROS的表达，减少IL-6的分泌。利用qRT-PCR和Western blot技术，我们发现，在POP患者的成纤维细胞中添加klotho可增加细胞生长，并以剂量依赖的方式降低IL-6和ROS的水平。结论：Klotho蛋白通过下调ERK1/2磷酸化，降低了POP患者成纤维细胞中MMP-1和MMP-3的表达。
Background: Pelvic organ prolapse (POP) is due to age-related atrophy and the weakening of the tissues of the pelvic floor, with degradation of collagen and extracellular matrix (ECM) by metalloproteinases (MMPs). This study aimed to investigates the role of the age-related enzyme klotho, encoded by the KL gene, in cultured fibroblasts obtained from patients with POP and the levels of reactive oxygen species (ROS), interleukin-6 (IL-6), and MMPs.Material/Methods: Pelvic floor fibroblasts were obtained from connective tissue from three patients with POP and three normal subjects. Cell proliferation and ROS production were measured using a cell counting kit-8 (CCK-8) assay and flow cytometry. Levels of interleukin-6 (IL-6), klotho, metalloproteinase-1 (MMP-1), MMP-3, extracellular signalre-gulated kinases 1/2 (ERK1/2), and p-ERK1/2 were measured by enzyme-linked immunoassay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot.Results: In cultured pelvic floor fibroblasts from patients with POP, the expression of klotho protein and klotho mRNA were significantly down-regulated in fibroblasts from patients with POP compared with normal fibroblasts. Klotho supplementation in cultured fibroblasts for patients with POP included increased cell growth, reduced expression of ROS reduction, and reduced the secretion of IL-6. Using qRT-PCR and Western blot, klotho supplementation of fibroblasts from patients with POP increased cell growth and reduced the levels of IL-6 and ROS in a dose-dependent way.Conclusions: Klotho protein reduced the expression of MMP-1 and MMP-3 in fibroblasts from patients with POP by down-regulating the phosphorylation of ERK1/2.