Jablonska Patrycja, Mierzejewska Paulina, Tomczyk Marta, Koszalka Patrycja, Franczak Marika, Kawecka Ada, Kutryb-Zajac Barbara, Braczko Alicja, Smolenski Ryszard T, Slominska Ewa M
Department of Biochemistry, Medical University of Gdansk, 80-211 Gdansk, Poland.
Institute of Medical Biotechnology and Experimental Oncology, Intercollegiate Faculty of Biotechnology of University of Gdansk and Medical University of Gdansk, Medical University of Gdansk, 80-211 Gdansk, Poland.
Biology (Basel). 2022 Apr 27;11(5):675. doi: 10.3390/biology11050675.
The disruption of the metabolism of extracellular NAD and NMN may affect related signaling cascades and pathologies, such as cardiovascular or respiratory system diseases. We aimed to study NAD and NMN hydrolysis on surface endothelial cells of diverse origins and with genetically modified nucleotide catabolism pathways. We tested lung endothelial cells isolated from C57BL/6 J wild-type (WT) and C57BL/6 J CD73 knockout (CD73 KO) mice, the transfected porcine iliac artery endothelial cell line (PIEC) with the human E5NT gene for CD73 (PIEC CD73), and a mock-transfected control (PIEC MOCK), as well as HMEC-1 and H5V cells. Substrate conversion into the product was followed by high-performance liquid chromatography (HPLC). We showed profound differences in extracellular NAD and NMN metabolism related to the vessel origin, species diversity, and type of culture. We also confirmed the involvement of CD38 and CD73 in NAD and NMN cleavage.
细胞外烟酰胺腺嘌呤二核苷酸（NAD）和烟酰胺单核苷酸（NMN）代谢的破坏可能会影响相关信号级联反应和病理状况，如心血管或呼吸系统疾病。我们旨在研究不同来源且具有基因修饰核苷酸分解代谢途径的表面内皮细胞上的NAD和NMN水解情况。我们测试了从C57BL / 6 J野生型（WT）和C57BL / 6 J CD73基因敲除（CD73 KO）小鼠分离的肺内皮细胞、转染了人E5NT基因以表达CD73的猪髂动脉内皮细胞系（PIEC CD73）、 mock转染对照（PIEC MOCK），以及人微血管内皮细胞系-1（HMEC-1）和人脐静脉内皮细胞系-5（H5V）细胞。通过高效液相色谱法（HPLC）追踪底物向产物的转化。我们发现细胞外NAD和NMN代谢在血管来源、物种多样性和培养类型方面存在显著差异。我们还证实了CD38和CD73参与NAD和NMN的裂解。
