Epigallocatechin-3-Gallate Promotes Recanalization in Deep Vein Thrombosis by Modulating Endothelial Progenitor Cell Ferroptosis Through the Nrf2 Pathway.

Abstract

Endothelial progenitor cells (EPCs) provide a promising therapeutic choice for deep venous thrombosis (DVT). Their number is increased by epigallocatechin-3-gallate (EGCG) in patients with diabetes. Although EGCG is effective against doxorubicin-induced ferroptosis and cardiotoxicity, its efficacy in DVT treatment has not been well studied. This study was aimed at assessing the effects of EGCG on EPC ferroptosis and recanalization in a DVT model. EPCs were treated with EGCG, and their proliferation and migration, angiogenesis, and apoptosis were evaluated using cell counting kit-8 and colony formation, Transwell, tube formation, and flow cytometry assays. Levels of iron, ferroptosis markers, and reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨm) were measured. Expression of ferroptosis-related genes and proteins was analyzed using qRT-PCR and western blotting, respectively. Promoter activation was evaluated using a dual-luciferase reporter system. Thrombus recanalization was examined in the DVT mouse model via hematoxylin and eosin staining and digital subtraction angiography. EGCG promoted EPC proliferation, migration, and angiogenesis and suppressed apoptosis. It attenuated ferroptosis by reducing iron and ROS accumulation, increasing ΔΨm, and regulating the expression of ferroptosis-related genes and proteins (ALOX15, ACSL4, and FTH1). EGCG enhanced the expression of Nrf2 and its targets, Slc7A11, HO-1, and GPX4. EGCG inhibited thrombogenesis and promoted recanalization in DVT mice, an effect mediated through the Nrf2 pathway and enhanced upon EPC transplantation. Transplantation of EGCG-pretreated EPCs facilitates DVT resolution via ferroptosis blockade. EGCG-pretreated EPC-based therapy may provide a novel option for patients with DVT.