Direct thrombin inhibitors fail to reverse the negative effects of heparin on lung growth and function after murine left pneumonectomy

• Published in: American journal of physiology. Lung cellular and molecular physiology Vol. 326; no. 3; pp. L213 - L225
• Main Authors: Tsikis, Savas T, Hirsch, Thomas I, Klouda, Timothy, Fligor, Scott C, Pan, Amy, Joiner, Malachi M, Wang, Sarah Z, Quigley, Mikayla, Devietro, Angela, Mitchell, Paul D, Kishikawa, Hiroko, Yuan, Ke, Puder, Mark
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.03.2024
• Subjects: Angiogenesis; Animal models; Animals; Anticoagulants; Anticoagulants - therapeutic use; Antithrombins - pharmacology; Antithrombins - therapeutic use; Apoptosis; Argatroban; Arginine - analogs & derivatives; Cell growth; Cell proliferation; Diaphragm; Disease Models, Animal; Endothelial cells; Fibrinolytic Agents; Gene expression; Heparin; Heparin - pharmacology; Heparin - therapeutic use; Hernia; Hirudins - pharmacology; Humans; In vivo methods and tests; Inhibitors; Lung - metabolism; Lungs; Mice; Mice, Inbred C57BL; Neonates; Peptide Fragments - pharmacology; Pipecolic Acids; Pneumonectomy; Pulmonary functions; Pumps; Recombinant Proteins - pharmacology; Respiratory function; Sulfonamides; Thrombin; Thrombin - metabolism; Thrombin - pharmacology; Vascularization; heparin; congenital diaphragmatic hernia; compensatory lung growth; pulmonary hypoplasia; anticoagulation
• ISSN: 1040-0605; 1522-1504; 1522-1504
• DOI: 10.1152/ajplung.00096.2023

Neonates with congenital diaphragmatic hernia (CDH) frequently require cardiopulmonary bypass and systemic anticoagulation. We previously demonstrated that even subtherapeutic heparin impairs lung growth and function in a murine model of compensatory lung growth (CLG). The direct thrombin inhibitors (DTIs) bivalirudin and argatroban preserved growth in this model. Although DTIs are increasingly used for systemic anticoagulation clinically, patients with CDH may still receive heparin. In this experiment, lung endothelial cell proliferation was assessed following treatment with heparin-alone or mixed with increasing concentrations of bivalirudin or argatroban. The effects of subtherapeutic heparin with or without DTIs in the CLG model were also investigated. C57BL/6J mice underwent left pneumonectomy and subcutaneous implantation of osmotic pumps. Pumps were preloaded with normal saline, bivalirudin, or argatroban; treated animals received daily intraperitoneal low-dose heparin. In vitro, heparin-alone decreased endothelial cell proliferation and increased apoptosis. The effect of heparin on proliferation, but not apoptosis, was reversed by the addition of bivalirudin and argatroban. In vivo, low-dose heparin decreased lung volume compared with saline-treated controls. All three groups that received heparin demonstrated decreased lung function on pulmonary function testing and impaired exercise performance on treadmill tolerance testing. These findings correlated with decreases in alveolarization, vascularization, angiogenic signaling, and gene expression in the heparin-exposed groups. Together, these data suggest that bivalirudin and argatroban fail to reverse the inhibitory effects of subtherapeutic heparin on lung growth and function. Clinical studies on the impact of low-dose heparin with DTIs on CDH outcomes are warranted. Infants with pulmonary hypoplasia frequently require cardiopulmonary bypass and systemic anticoagulation. We investigate the effects of simultaneous exposure to heparin and direct thrombin inhibitors (DTIs) on lung growth and pulmonary function in a murine model of compensatory lung growth (CGL). Our data suggest that DTIs fail to reverse the inhibitory effects of subtherapeutic heparin on lung growth and function. Clinical studies on the impact of heparin with DTIs on clinical outcomes are thus warranted.