Gdf11 Overexpression Alleviates Sepsis-Induced Lung Microvascular Endothelial Barrier Damage by Activating Sirt1/Nox4 Signaling to Inhibit Ferroptosis

Abstract

Sepsis is a lethal clinical syndrome, and acute lung injury (ALI) is the earliest and most serious complication. We aimed to explore the role of growth differentiation factor 11 (GDF11) in sepsis-induced dysfunction of lung microvascular endothelial barrier in vivo and in vitro to elucidate its potential mechanism related to sirtuin 1 (SIRT1)/NADPH oxidase 4 (NOX4) signaling. Cecal ligation and puncture (CLP)-induced sepsis mice and lipopolysaccharide (LPS)-induced pulmonary microvascular endothelial cells (PMECs) were use in this study. Histopathological changes in lung tissues were tested by H&E staining. Lung wet-to-dry weight ratio and inflammatory factors contents in bronchoalveolar lavage fluid (BALF) were assessed. Evens blue index, TEER and expression of zona occludens 1 (ZO-1), occludin-1 and Claudin-1 were used to evaluate alveolar barrier integrity. Reactive oxygen species (ROS), lipid peroxidation and ferroptosis markers were analyzed. Iron deposition in the lung tissues was assessed using Prussian blue staining. Intracellular Fe2+ level was detected using FerroOrange staining. Additionally, expression of GDF11, SIRT1 and NOX4 was estimated with western blot. Then, EX527, a SIRT1 inhibitor, was employed to treat GDF11-overexpressed PMECs with LPS stimulation to clarify the regulatory mechanism. Results showed that GDF11 overexpression attenuated sepsis-induced pathological changes and inflammation and maintained alveolar barrier integrity. Moreover, GDF11 overexpression inhibited ferroptosis, upregulated SIRT1 expression and downregulated NOX4 expression. Additionally, EX527 treatment relieved the impacts of GDF11 overexpression on ferroptosis and destruction of integrity of HPMVECs exposed to LPS. Taken together, GDF11 overexpression could alleviate sepsis-induced lung microvascular endothelial barrier damage by activating SIRT1/NOX4 signaling to inhibit ferroptosis. Our findings potentially provide new molecular target for clinical therapy of ALI.