Silencing of LOX-1 attenuates high glucose-induced ferroptosis in THVECs via the HIF-1α/SLC7A11 signaling pathway

Abstract

Objectives

Diabetic osteoporosis (DOP) represents a significant and serious complication associated with diabetes, characterized by a complex and inadequately understood pathophysiological mechanism. Recent studies have highlighted a robust association between DOP and ferroptosis. H-type vessels play a critical role in osteoporosis, while lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is associated with endothelial dysfunction related to diabetes. In this study, we investigate how LOX-1 affects ferroptosis in H-type vascular endothelial cells (THVECs) under high glucose (HG) conditions, aiming to elucidate the molecular mechanisms involved.

Methods

THVECs were isolated from rats employing an enzymatic digestion method and subsequently validated through immunofluorescence analysis. The silencing of LOX-1 was achieved via transfection with a lentiviral vector. Cell viability was assessed using the CCK-8 assay, and ROS, MMP, GSH, MDA, and Fe²⁺ levels were assessed utilizing specific commercial kits. Western blotting and PCR assessed LOX-1, HIF-1α, SLC7A11, and GPX4 expression levels.

Results

In high glucose conditions, LOX-1 expression at both protein and mRNA levels increased, while ROS, MDA, and Fe²⁺ rose, and MMP and GSH levels fell, resulting in ferroptosis in THVECs. This condition could be reversed by silencing LOX-1 or by administering the ferroptosis inhibitor (Fer-1). Further analysis showed that silencing LOX-1 enhanced the expression of HIF-1α, SLC7A11, and GPX4, which mitigated ferroptosis in THVECs.

Conclusions

Downregulation of LOX-1 alleviates high glucose-induced ferroptosis in THVECs via the HIF-1α/SLC7A11 pathway. This suggests that LOX-1 functions as a critical target for regulating ferroptosis in THVECs, providing a novel insight into the pathological mechanisms associated with DOP.