LncRNA MALAT1 facilitates BM-MSCs differentiation into endothelial cells and ameliorates erectile dysfunction via the miR-206/CDC42/PAK1/paxillin signalling axis.

BACKGROUND: Erectile dysfunction (ED) is a common male sexual dysfunction, with an increasing incidence, and the current treatment is often ineffective. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Pseudomonas zeshuii sp. nov., isolated from herbicide-contaminated soil.

A Gram-stain-negative, rod-shaped, non-motile, non-spore-forming bacterium, designated strain BY-1(T), was isolated from a soil sample from the city of Qiqihar in Heilongjiang Province, PR China. Strain BY-1(T) grew optimally at pH 7.0 and 30-35 °C in the presence of 0.5% (w/v) NaCl. Analysis of 16S rRNA gene sequences revealed that strain BY-1(T) fell within the radiation of the genus Pseudomonas, and showed highest 16S rRNA gene sequence similarities to Pseudomonas luteola IAM 13000(T) (99.5%) and Pseudomonas duriflava HR2(T) (97.3%); the levels of sequence similarity with respect to other recognized species of the genus Pseudomonas were <96.7%. Strain BY-1(T) showed low DNA-DNA relatedness values with Pseudomonas luteola IAM 13000(T) (29 ± 3.1%) and Pseudomonas duriflava HR2(T) (21 ± 1.5 %). The G+C content of the genomic DNA of strain BY-1(T) was 55.3 mol%. The major fatty acids were C(18:1), C(16:0) and summed feature 3 (C(16:1)ω6c and/or C(16:1)ω7c). Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine, and the major ubiquinone was Q-9. Data obtained in this study indicated that this isolate represents a novel species of the genus Pseudomonas, for which the name Pseudomonas zeshuii sp. nov. is proposed. The type strain is BY-1(T) (=KACC 15471(T)=ACCC 05688(T)).