Rybakowska Iwona, Romaszko Pawel, Zabielska Magdalena, Turyn Jacek, Kaletha Krystian, Barton Paul J, Slominska Ewa M, Smolenski Ryszard T
a Department of Biochemistry and Clinical Physiology , Medical University of Gdansk , Gdansk , Poland.
Nucleosides Nucleotides Nucleic Acids. 2014;33(4-6):319-22. doi: 10.1080/15257770.2014.880481.
Recent findings suggest that inhibition of AMP-deaminase (AMPD) could be effective therapeutic strategy in heart disease associated with cardiac ischemia. To establish experimental model to study protective mechanisms of AMPD inhibition we developed conditional, cardiac specific knock-outs in Cre recombinase system. AMPD3 floxed mice were crossed with Mer-Cre-Mer mice. Tamoxifen was injected to induce Cre recombinase. After two weeks, hearts, skeletal muscle, liver, kidney, and blood were collected and activities of AMPD and related enzymes were analyzed using HPLC-based procedure. We demonstrate loss of more than 90% of cardiac AMPD activity in the heart of AMPD3-/-mice while other enzymes of nucleotide metabolism such as adenosine deaminase, purine nucleoside phosphorylase were not affected. Surprisingly, activity of AMPD was also reduced in the erythrocytes and in the kidney by 20%-30%. No change of AMPD activity was observed in the skeletal muscle and the liver.
最近的研究结果表明，抑制AMP脱氨酶（AMPD）可能是治疗与心肌缺血相关心脏病的有效策略。为了建立实验模型来研究AMPD抑制的保护机制，我们在Cre重组酶系统中开发了条件性、心脏特异性基因敲除小鼠。将AMPD3基因floxed小鼠与Mer-Cre-Mer小鼠杂交。注射他莫昔芬以诱导Cre重组酶。两周后，收集心脏、骨骼肌、肝脏、肾脏和血液，并使用基于高效液相色谱的方法分析AMPD和相关酶的活性。我们证明，在AMPD3-/-小鼠的心脏中，心脏AMPD活性丧失超过90%，而核苷酸代谢的其他酶，如腺苷脱氨酶、嘌呤核苷磷酸化酶不受影响。令人惊讶的是，红细胞和肾脏中的AMPD活性也降低了20%-30%。在骨骼肌和肝脏中未观察到AMPD活性的变化。
