Department of Clinical Infection Microbiology and Immunology, University of Liverpool, Liverpool L69 7BE, UK.
Coagulation Department, Liverpool University Hospitals NHS Foundation Trust, Liverpool L7 8XP, UK.
Int J Mol Sci. 2023 Feb 6;24(4):3225. doi: 10.3390/ijms24043225.
Reduction in cardiac contractility is common in severe sepsis. However, the pathological mechanism is still not fully understood. Recently it has been found that circulating histones released after extensive immune cell death play important roles in multiple organ injury and disfunction, particularly in cardiomyocyte injury and contractility reduction. How extracellular histones cause cardiac contractility depression is still not fully clear. In this work, using cultured cardiomyocytes and a histone infusion mouse model, we demonstrate that clinically relevant histone concentrations cause significant increases in intracellular calcium concentrations with subsequent activation and enriched localization of calcium-dependent protein kinase C (PKC) α and βII into the myofilament fraction of cardiomyocytes in vitro and in vivo. Furthermore, histones induced dose-dependent phosphorylation of cardiac troponin I (cTnI) at the PKC-regulated phosphorylation residues (S43 and T144) in cultured cardiomyocytes, which was also confirmed in murine cardiomyocytes following intravenous histone injection. Specific inhibitors against PKCα and PKCβII revealed that histone-induced cTnI phosphorylation was mainly mediated by PKCα activation, but not PKCβII. Blocking PKCα also significantly abrogated histone-induced deterioration in peak shortening, duration and the velocity of shortening, and re-lengthening of cardiomyocyte contractility. These in vitro and in vivo findings collectively indicate a potential mechanism of histone-induced cardiomyocyte dysfunction driven by PKCα activation with subsequent enhanced phosphorylation of cTnI. These findings also indicate a potential mechanism of clinical cardiac dysfunction in sepsis and other critical illnesses with high levels of circulating histones, which holds the potential translational benefit to these patients by targeting circulating histones and downstream pathways.
心肌收缩力降低在严重脓毒症中很常见。然而,其病理机制仍不完全清楚。最近发现,广泛的免疫细胞死亡后释放的循环组蛋白在多个器官损伤和功能障碍中发挥重要作用,特别是在心肌细胞损伤和收缩力降低中。细胞外组蛋白如何引起心肌收缩力降低仍不完全清楚。在这项工作中,我们使用培养的心肌细胞和组蛋白输注小鼠模型,证明临床相关浓度的组蛋白会导致细胞内钙离子浓度显著增加,随后钙离子依赖性蛋白激酶 C(PKC)α和βII在体外和体内的心肌细胞肌丝部分被激活并富集定位。此外,组蛋白诱导培养的心肌细胞中心肌肌钙蛋白 I(cTnI)在 PKC 调节的磷酸化残基(S43 和 T144)处发生剂量依赖性磷酸化,静脉注射组蛋白后也在小鼠心肌细胞中得到证实。PKCα 和 PKCβII 的特异性抑制剂表明,组蛋白诱导的 cTnI 磷酸化主要是通过 PKCα 的激活介导的,而不是 PKCβII。阻断 PKCα 还显著阻断了组蛋白诱导的心肌细胞收缩力峰值缩短、持续时间和缩短速度以及再伸长的恶化。这些体内外研究结果共同表明,组蛋白通过激活 PKCα 诱导心肌细胞功能障碍的潜在机制,随后 cTnI 的磷酸化增强。这些发现还表明,在循环组蛋白水平较高的脓毒症和其他危重病中,临床心脏功能障碍的潜在机制,通过靶向循环组蛋白和下游途径,为这些患者带来潜在的转化益处。