Jiangxi Provincial Key Laboratory of Urinary System Diseases, Department of Urology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, No. 17, Yongwai Zheng Street, Nanchang, 330006, China.
Nanchang University, No. 999 Xuefu Avenue, Honggutan District, Nanchang City, 330006, Jiangxi Province, China.
Reprod Biol Endocrinol. 2024 Jun 25;22(1):74. doi: 10.1186/s12958-024-01240-8.
Erectile dysfunction (ED) is a common male sexual dysfunction, with an increasing incidence, and the current treatment is often ineffective.
Vascular endothelial growth factor (VEGFA) was used to treat bone marrow-derived mesenchymal stem cells (BM-MSCs), and their cell migration rates were determined by Transwell assays. The expression of the von Willebrand Factor (vWF)VE-cadherin, and endothelial nitric oxide synthase(eNOS) endothelial markers was determined by qRT‒PCR and Western blot analyses. The MALAT1-induced differentiation of BM-MCs to ECs via the CDC42/PAK1/paxillin pathway was explored by transfecting VEGFA-induced BM-MSC with si-MALAT1 and overexpressing CDC42 and PAK1. The binding capacity between CDC42, PAK1, and paxillin in VEGFA-treated and non-VEGFA-treated BM-MSCs was examined by protein immunoprecipitation. MiR-206 was overexpressed in VEGFA-induced BM-MSC, and the binding sites of MALAT1, miR-206, and CDC42 were identified using a luciferase assay. Sixty male Sprague‒Dawley rats were divided into six groups (n = 10/group). DMED modelling was demonstrated by APO experiments and was assessed by measuring blood glucose levels. Erectile function was assessed by measuring the intracavernosa pressure (ICP) and mean arterial pressure (MAP). Penile erectile tissue was analysed by qRT‒PCR, Western blot analysis, and immunohistochemical staining.
MALAT1 under VEGFA treatment conditions regulates the differentiation of BM-MSCs into ECs by modulating the CDC42/PAK1/paxillin axis. In vitro experiments demonstrated that interference with CDC42 and MALAT1 expression inhibited the differentiation of BM-MSCs to ECs. CDC42 binds to PAK1, and PAK1 binds to paxillin. In addition, CDC42 in the VEGFA group had a greater ability to bind to PAK1, whereas PAK1 in the VEGFA group had a greater ability to bind to paxillin. Overexpression of miR-206 in VEGFA-induced BM-MSCs demonstrated that MALAT1 competes with the CDC42 3'-UTR for binding to miR-206, which in turn is involved in the differentiation of BM-MSCs to ECs. Compared to the DMED model group, the ICP/MAP ratio was significantly greater in the three BM-MSCs treatment groups.
MALAT1 facilitates BM-MSC differentiation into ECs by regulating the miR-206/CDC42/PAK1/paxillin axis to improve ED. The present findings revealed the vital role of MALAT1 in the repair of BM-MSCs for erectile function and provided new mechanistic insights into the BM-MSC-mediated repair of DMED.
勃起功能障碍（ED）是一种常见的男性性功能障碍，发病率不断增加，目前的治疗方法往往效果不佳。
使用血管内皮生长因子（VEGFA）处理骨髓间充质干细胞（BM-MSCs），并通过 Transwell 测定其细胞迁移率。通过 qRT-PCR 和 Western blot 分析测定血管性血友病因子（vWF）VE-钙粘蛋白和内皮型一氧化氮合酶（eNOS）内皮标志物的表达。通过转染 VEGFA 诱导的 BM-MSC 中的 si-MALAT1 和过表达 CDC42 和 PAK1，探讨 MALAT1 通过 CDC42/PAK1/paxillin 途径诱导 BM-MCs 向 ECs 分化。通过蛋白质免疫沉淀检测 VEGFA 处理和未处理的 BM-MSCs 中 CDC42、PAK1 和 paxillin 之间的结合能力。在 VEGFA 诱导的 BM-MSC 中转染 miR-206 过表达，通过荧光素酶测定鉴定 MALAT1、miR-206 和 CDC42 的结合位点。60 只雄性 Sprague-Dawley 大鼠分为 6 组（n=10/组）。通过 APO 实验证实 DMED 模型，并通过测量血糖水平进行评估。通过测量海绵体压（ICP）和平均动脉压（MAP）评估阴茎勃起功能。通过 qRT-PCR、Western blot 分析和免疫组织化学染色分析阴茎勃起组织。
MALAT1 在 VEGFA 处理条件下通过调节 CDC42/PAK1/paxillin 轴调节 BM-MSCs 向 ECs 的分化。体外实验表明，干扰 CDC42 和 MALAT1 的表达抑制了 BM-MSCs 向 ECs 的分化。CDC42 与 PAK1 结合，PAK1 与 paxillin 结合。此外，VEGFA 组中的 CDC42 与 PAK1 结合的能力更强，而 VEGFA 组中的 PAK1 与 paxillin 结合的能力更强。在 VEGFA 诱导的 BM-MSCs 中转染 miR-206 过表达表明，MALAT1 与 CDC42 3'-UTR 竞争结合 miR-206，从而参与 BM-MSCs 向 ECs 的分化。与 DMED 模型组相比，三个 BM-MSCs 治疗组的 ICP/MAP 比值显著增大。
MALAT1 通过调节 miR-206/CDC42/PAK1/paxillin 轴促进 BM-MSC 向 ECs 分化，从而改善 ED。本研究结果揭示了 MALAT1 在 BM-MSCs 修复勃起功能中的重要作用，并为 BM-MSC 介导的 DMED 修复提供了新的机制见解。
