Department of Morphology, Surgery and Experimental Medicine, Pharmacology Section, University of Ferrara, 44121 Ferrara, Italy.
Department of Biomedical and Specialist Surgical Sciences, Section of Neurological, Psychiatric and Psychological Sciences, University of Ferrara, 44121 Ferrara, Italy.
Cells. 2020 Apr 25;9(5):1072. doi: 10.3390/cells9051072.
Adenosine modulates many physiological processes through the interaction with adenosine receptors (ARs) named as A, A, A and AARs. During ischemic stroke, adenosine mediates neuroprotective and anti-inflammatory effects through ARs activation. One of the dominant pathways generating extracellular adenosine involves the dephosphorylation of ATP by ecto-nucleotidases CD39 and CD73, which efficiently hydrolyze extracellular ATP to adenosine. The aim of the study is to assess the presence of ARs in lymphocytes from ischemic stroke patients compared to healthy subjects and to analyze changes in CD39 and CD73 expression in CD4 and CD8 lymphocytes. Saturation binding experiments revealed that AARs affinity and density were significantly increased in ischemic stroke patients whilst no differences were found in A, A and AARs. These results were also confirmed in reverse transcription (RT)-polymerase chain reaction (PCR) assays where AAR mRNA levels of ischemic stroke patients were higher than in control subjects. In flow cytometry experiments, the percentage of CD73 cells was significantly decreased in lymphocytes and in T-lymphocyte subclasses CD4 and CD8 obtained from ischemic stroke patients in comparison with healthy individuals. These data corroborate the importance of the adenosinergic system in ischemic stroke and could open the way to more targeted therapeutic approaches and biomarker development for ischemic stroke.
腺苷通过与命名为 A、A、A 和 AAR 的腺苷受体(AR)相互作用,调节许多生理过程。在缺血性中风中,通过 AR 激活,腺苷介导神经保护和抗炎作用。产生细胞外腺苷的主要途径之一涉及细胞外核苷酸酶 CD39 和 CD73 将 ATP 去磷酸化,这两种酶有效地将细胞外 ATP 水解为腺苷。本研究旨在评估与健康受试者相比,缺血性中风患者淋巴细胞中 AR 的存在,并分析 CD4 和 CD8 淋巴细胞中 CD39 和 CD73 表达的变化。饱和结合实验表明,缺血性中风患者的 AAR 亲和力和密度显著增加,而 A、A 和 AAR 则没有差异。逆转录(RT)-聚合酶链反应(PCR)检测也证实了这一结果,其中缺血性中风患者的 AAR mRNA 水平高于对照组。在流式细胞术实验中,与健康个体相比,从缺血性中风患者获得的淋巴细胞和 T 淋巴细胞亚群 CD4 和 CD8 中 CD73 细胞的百分比显著降低。这些数据证实了腺苷能系统在缺血性中风中的重要性,并为缺血性中风的更有针对性的治疗方法和生物标志物的发展开辟了道路。