Departamento de Ciências Básicas da Saúde and Laboratório de Biologia Celular, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Porto Alegre, RS, Brazil.
Laboratório de Sinalização e Plasticidade Celular, Departamento de Biofísica, Instituto de Biociências UFRGS, Porto Alegre, RS, Brazil.
Life Sci. 2020 Sep 1;256:117862. doi: 10.1016/j.lfs.2020.117862. Epub 2020 May 28.
Vascular smooth muscle cells (VSMCs) exhibit a high degree of plasticity when they undergo the progression from a normal to a disease condition, which makes them a potential target for evaluating early markers and for the development of new therapies. Purinergic signalling plays a key role in vascular tonus control, ATP being an inductor of vasoconstriction, whereas adenosine mediates a vasodilation effect antagonising the ATP actions. The control of extracellular ATP and adenosine levels is done by ectonucleotidases, which represent a potential target to be evaluated in the progression of cardiovascular diseases. In this study, we analysed the basal activity and expression of the ectonucleotidases in aortic rat VSMCs, and we further performed in silico analysis to determine the expression of those enzymes in conditions that mimicked vascular diseases. Cultured in vitro VSMCs showed a prominent expression of Entpd1 followed by Entpd2 and Nt5e (CD73) and very low levels of Entpd3. Slightly faster AMP hydrolysis was observed when compared to ATP and ADP nucleotides. In silico analysis showed that the ectonucleotidases were modulated after induction of conditions that can lead to vascular diseases such as, hypertensive and hypotensive mice models (Nt5e); exposition to high-fat (Entpd1 and Entpd2) or high-phosphate (Nt5e) diet; mechanical stretch (Entpd1, Entpd2 and Nt5e); and myocardial infarction (Entpd1). Our data show that VSMCs are able to efficiently metabolise the extracellular nucleotides generating adenosine. The modulation of Entpd1, Entdp2 and Nt5e in vascular diseases suggests these ectoenzymes as potential targets or markers to be investigated in future studies.
血管平滑肌细胞（VSMCs）在从正常状态向疾病状态发展时表现出高度的可塑性，这使它们成为评估早期标志物和开发新疗法的潜在靶点。嘌呤能信号在血管张力控制中起着关键作用，ATP 是血管收缩的诱导剂，而腺苷则介导血管舒张作用，拮抗 ATP 的作用。细胞外 ATP 和腺苷水平的控制是通过核苷酸酶完成的，核苷酸酶是评估心血管疾病进展的潜在靶点。在这项研究中，我们分析了大鼠主动脉 VSMCs 中核苷酸酶的基础活性和表达，并进一步进行了计算机模拟分析，以确定在模拟血管疾病的条件下这些酶的表达情况。体外培养的 VSMCs 表现出 Entpd1 的显著表达，其次是 Entpd2 和 Nt5e（CD73），Entpd3 的表达水平非常低。与 ATP 和 ADP 核苷酸相比，AMP 的水解速度稍快。计算机模拟分析表明，在诱导可能导致血管疾病的条件后，核苷酸酶被调节，例如高血压和低血压小鼠模型（Nt5e）；暴露于高脂肪（Entpd1 和 Entpd2）或高磷（Nt5e）饮食；机械拉伸（Entpd1、Entpd2 和 Nt5e）；和心肌梗死（Entpd1）。我们的数据表明，VSMCs 能够有效地代谢细胞外核苷酸生成腺苷。在血管疾病中 Entpd1、Entdp2 和 Nt5e 的调节表明这些外切酶作为潜在的靶点或标志物有待进一步研究。
