Department of Surgery University of Virginia Charlottesville VA.
Department of Pharmacology Changzhi Medical College Changzhi City Shanxi Province China.
J Am Heart Assoc. 2021 Aug 3;10(15):e020754. doi: 10.1161/JAHA.121.020754. Epub 2021 Jul 30.
Background We previously demonstrated that ischemically injured cardiomyocytes release cell-free DNA and HMGB1 (high mobility group box 1 protein) into circulation during reperfusion, activating proinflammatory responses and ultimately exacerbating reperfusion injury. We hypothesize that cell-free DNA and HMGB1 mediate myocardial ischemia-reperfusion injury by stimulating plasmacytoid dendritic cells (pDCs) to secrete type I interferon (IFN-I). Methods and Results C57BL/6 and interferon alpha receptor-1 knockout mice underwent 40 minutes of left coronary artery occlusion followed by 60 minutes of reperfusion (40'/60' IR) before infarct size was evaluated by 2,3,5-Triphenyltetrazolium chloride-Blue staining. Cardiac perfusate was acquired in ischemic hearts without reperfusion by antegrade perfusion of the isolated heart. Flow cytometry in pDC-depleted mice treated with multiple doses of plasmacytoid dendritic cell antigen-1 antibody via intraperitoneal injection demonstrated plasmacytoid dendritic cell antigen-1 antibody treatment had no effect on conventional splenic dendritic cells but significantly reduced splenic pDCs by 60%. pDC-depleted mice had significantly smaller infarct size and decreased plasma interferon-α and interferon-β compared with control. Blockade of the type I interferon signaling pathway with cyclic GMP-AMP synthase inhibitor, stimulator of interferon genes antibody, or interferon regulatory factor 3 antibody upon reperfusion similarly significantly attenuated infarct size by 45%. Plasma levels of interferon-α and interferon-β were significantly reduced in cyclic GMP-AMP synthase inhibitor-treated mice. Infarct size was significantly reduced by >30% in type I interferon receptor monoclonal antibody-treated mice and interferon alpha receptor-1 knockout mice. In splenocyte culture, 40'/0' cardiac perfusate treatment stimulated interferon-α and interferon-β production; however, this effect disappeared in the presence of cyclic GMP-AMP synthase inhibitor. Conclusions Type I interferon production is stimulated following myocardial ischemia by cardiogenic cell-free DNA/HMGB1 in a pDC-dependent manner, and subsequently activates type I interferon receptors to exacerbate reperfusion injury. These results identify new potential therapeutic targets to attenuate myocardial ischemia-reperfusion injury.
我们之前的研究表明,在再灌注期间,缺血性损伤的心肌细胞将无细胞 DNA 和高迁移率族蛋白 B1(HMGB1)释放到循环中,激活促炎反应,最终加重再灌注损伤。我们假设无细胞 DNA 和 HMGB1 通过刺激浆细胞样树突状细胞(pDC)分泌 I 型干扰素(IFN-I)来介导心肌缺血再灌注损伤。
C57BL/6 和干扰素-α受体-1 敲除小鼠进行 40 分钟左冠状动脉结扎,随后进行 60 分钟再灌注(40'/60'IR),然后通过 2,3,5-三苯基四氮唑蓝染色评估梗死面积。在未再灌注的缺血心脏中通过逆行心脏灌注获得心脏灌流液。通过腹腔注射多次剂量的浆细胞样树突状细胞抗原-1 抗体处理 pDC 耗竭小鼠的流式细胞术显示,浆细胞样树突状细胞抗原-1 抗体处理对常规脾脏树突状细胞没有影响,但可使脾脏 pDC 减少 60%。pDC 耗竭小鼠的梗死面积明显较小,且血浆干扰素-α和干扰素-β水平降低。再灌注时用环鸟苷酸-腺苷酸合酶抑制剂、干扰素基因刺激剂抗体或干扰素调节因子 3 抗体阻断 I 型干扰素信号通路同样可使梗死面积显著减少 45%。环鸟苷酸-腺苷酸合酶抑制剂治疗小鼠的血浆干扰素-α和干扰素-β水平显著降低。I 型干扰素受体单克隆抗体处理和干扰素-α受体-1 敲除小鼠的梗死面积分别减少 30%以上。在脾细胞培养中,40'/0'心脏灌流液处理刺激了干扰素-α和干扰素-β的产生;然而,在存在环鸟苷酸-腺苷酸合酶抑制剂的情况下,这种作用消失了。
心肌缺血后,心脏来源的无细胞 DNA/HMGB1 以 pDC 依赖性方式刺激 I 型干扰素产生,随后激活 I 型干扰素受体,加重再灌注损伤。这些结果确定了新的潜在治疗靶点,以减轻心肌缺血再灌注损伤。