Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine-University, Dusseldorf, Germany.
Department of Molecular Cardiology, Medical Faculty, Heinrich-Heine-University, Dusseldorf, Germany.
FASEB J. 2018 Aug;32(8):4356-4369. doi: 10.1096/fj.201701326R. Epub 2018 Mar 20.
Calcific aortic valve disease is an active disease process with lipoprotein deposition, chronic inflammation, and progressive leaflet degeneration. Expression of ectonucleotidases, a group of membrane-bound enzymes that regulate the metabolism of ATP and its metabolites, may coregulate the degeneration process of valvular interstitial cells (VICs). The aim of this study was to investigate the role of the enzymes of the purinergic system in the degeneration process of VICs. Ovine VICs were cultivated in vitro under different prodegenerative conditions and treated with inhibitors of ectonucleoside triphosphate diphosphohydrolase 1 (CD39)/ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), and 5'-nucleotidase (CD73), as well as with adenosine and adenosine receptor agonists. Experiments were performed both in 2-dimensional (2-D) and 3-dimensional (3-D) cell-culture models. Our main findings were that VICs continuously release ATP. Inhibition of ATP hydrolyzing enzymes (CD39 and ENPP1) resulted in profound prodegenerative effects with a vigorous up-regulation of CD39, ENPP1, and CD73, as well as TGF-β1 and osteopontin at the gene level. In our 3-D model, the effect was more pronounced than in 2-D monolayers. Increasing adenosine levels, as well as stimulating the adenosine receptors A and A, exhibited strong prodegenerative effects, whereas conversely, lowering adenosine levels by inhibition of CD73 resulted in protective effects against degeneration. Dysregulation of any one of these enzymes plays an important role in the degeneration process of VICs. Stimulation of ATP and adenosine has prodegenerative effects, whereas lowering the adenosine levels exerts a protective effect.-Weber, A., Barth, M., Selig, J. I., Raschke, S., Dakaras, K., Hof, A., Hesse, J., Schrader, J., Lichtenberg, A., Akhyari, P. Enzymes of the purinergic signaling system exhibit diverse effects on the degeneration of valvular interstitial cells in a 3-D microenvironment.
钙化性主动脉瓣疾病是一种脂蛋白沉积、慢性炎症和进行性瓣叶退行性变的主动疾病过程。细胞外核苷酸酶（一组调节 ATP 及其代谢物代谢的膜结合酶）的表达可能共同调节瓣膜间质细胞（VIC）的退行性变过程。本研究旨在探讨嘌呤能系统的酶在 VIC 退行性变过程中的作用。在不同的退行性变条件下，体外培养绵羊 VIC，并使用细胞外核苷酸三磷酸二磷酸水解酶 1（CD39）/核苷酸焦磷酸酶/磷酸二酯酶 1（ENPP1）和 5'-核苷酸酶（CD73）的抑制剂以及腺苷和腺苷受体激动剂进行处理。在 2 维（2-D）和 3 维（3-D）细胞培养模型中进行了实验。我们的主要发现是 VIC 持续释放 ATP。抑制 ATP 水解酶（CD39 和 ENPP1）会导致强烈的退行性变效应，基因水平上 CD39、ENPP1 和 CD73 以及 TGF-β1 和骨桥蛋白的上调非常明显。在我们的 3-D 模型中，这种效应比 2-D 单层更为明显。增加腺苷水平以及刺激腺苷受体 A 和 A2 表现出强烈的退行性变作用，而相反，通过抑制 CD73 降低腺苷水平则对退行性变具有保护作用。这些酶中的任何一种失调都会在 VIC 的退行性变过程中发挥重要作用。刺激 ATP 和腺苷具有退行性变作用，而降低腺苷水平则具有保护作用。-Weber，A.，Barth，M.，Selig，J. I.，Raschke，S.，Dakaras，K.，Hof，A.，Hesse，J.，Schrader，J.，Lichtenberg，A.，Akhyari，P. 嘌呤能信号系统的酶在 3-D 微环境中对瓣膜间质细胞的退行性变表现出不同的影响。
