HMOX1 upregulation promotes ferroptosis in diabetic atherosclerosis
Introduction
Cardiovascular diseases (CVDs) are the leading causes of morbidity and mortality worldwide [1]. Reducing atherosclerotic cardiovascular disease is a major clinical imperative. However, when the risks of atherosclerosis concordance with diabetes, the development of atherosclerosis is accelerated and more diffusely distributed than in individuals without diabetes [2], [3], [4]. Metabolic disorders are closely involved in the pathogenesis of atherosclerosis [5], especially in the arterial wall and front line endothelial cells (ECs). To explore an effective treatment strategy for diabetic atherosclerosis, it is important to dissect pathogenic mechanisms and identify meaningful targets.
Accelerated diabetic atherosclerosis occurs when oxidized low-density lipoproteins (oxLDLs) or oxidized lipids trapped in the vessel wall induce the overlying ECs to express adhesion molecules and cytokines, promoting the recruitment of monocytes and lymphocytes to the vessel wall [6], [7]. Although oxidized 1-palmitoyl-2-arachidonoyl-snglycero-3-phosphatidylcholine (Ox-PAPC, a component in atherosclerotic lesions [8]) is a critical factor in atherosclerosis [9], it is also a key factor associated with diabetic atherosclerosis development [10], [11]. However, the precise pathological alterations in the response of ECs to diabetic risk factors and the underlying mechanism are still poorly understood. Hence, we attempted to clarify the genetic characteristics and signaling pathways to study how atherosclerosis affects front-line ECs in human samples and then determine whether the identified genes are involved in diabetes. Completing these experiments will be helpful to enhance our understanding of diabetic atherosclerosis and to identify novel interventions preventing diabetic atherosclerosis.
We built our investigation on gene microarray technology (mRNA expression profiles), which has been widely applied in recent decades [12], [13]. In recent years, numerous studies on the gene expression profile of atherosclerosis have revealed hundreds of differentially expressed genes (DEGs), which are the basis for gene regulatory network analysis [13], [14] and avoid limitations or inconsistencies due to tissue or sample heterogeneity. In this study, we conducted in-depth analysis of two microarray datasets, GSE30169 [15] and GSE6584 [16], from the Gene Expression Omnibus database (GEO) [7] in an attempt to identify the candidate genes and pathological pathways that may contribute to diabetic atherosclerotic endothelial cell injury and could be used to prevent diabetic atherosclerosis. Moreover, the causative roles of genes identified from these databases were investigated by in vitro and in vivo diabetic animal models.
Therefore, the objectives of this study were first to identify the EC injury mode in the development of diabetic atherosclerosis and second to investigate the candidate genes and the mechanisms responsible for the development of diabetic atherosclerosis.
Section snippets
Screening of DEGs and performing pathway enrichment analysis
The GSE30169 and GSE6584 gene expression profiles and their relevant platform annotation files were obtained from the GEO database. The GSE30169 dataset was submitted by Professors Romanoski and Lusis on June 23, 2011, last updated on January 17, 2017, and stockpiled on the GPL3921 platform (HT_HG-U133A) Affymetrix HT Human Genome U133A Array (Affymetrix; Thermo Fisher Scientific, Inc., Waltham, MA, USA). [17] GSE30169 consisted of 629 samples from primary human aortic endothelial cells (HAECs)
Bioinformatic assays of human atherosclerotic vascular injury identified ferroptosis as an important form of cell death
To identify potential novel mechanisms contributing to atherosclerotic vascular injury in a nonbiased fashion, we first gathered information from a human bioinformatic database to identify the major form of cell death in the human atherosclerotic vascular injury database (gene expression profiles of GSE30169 and GSE6584 obtained from the NCBI-GEO free database). With pĀ <Ā 0.05 and [log2 FC]Ā ā„Ā 0.6 as cutoff criteria, we extracted 166 and 2604 differentially expressed genes (DEGs), respectively,
Discussion
In the present study, we reported that ferroptosis contributes to the progression of diabetic atherosclerosis. The increase in HMOX1 contributes to endothelial cell ferroptosis by promoting iron overload, ROS generation and lipid peroxide (Fig. 8). A diabetic increase in HMOX-1 indicates that HMOX-1 is a novel marker for diabetic endothelial dysfunction.
Atherosclerosis is the pathological basis of diabetic macrovascular complications, and it can evaluate the severity of macroangiopathy in T2DM.
Conclusions
Utilizing nonbiased in silico analysis followed by in vitro molecular investigation and in vivo concept-proven demonstration, we demonstrate that HMOX1 upregulation and resultant ferroptosis are involved in diabetic atherosclerosis. These results suggest that interventions blocking ferroptosis, such as Fer-1 administration and genetic/pharmacologic HMOX1 inhibition, may be novel approaches to attenuate diabetic atherosclerosis.
The following is the supplementary data related to this article.
Declaration of competing interest
The authors have declared that no competing interest exists.
Acknowledgements
This work was supported by the Scientific Research Project of the Shanxi Provincial Department of Health (Grant No. 2018023); Natural Science Foundation of China (81700327, 81970391, 82000799); Shanxi ā1331 Projectā Key Subjects Construction (1331KSC); Shanxi āSanjin Scholarsā Program; Key Laboratory of Cellular Physiology (Shanxi Medical University); Applied Basic Research Program of Shanxi Province (201801D221269); Scientific and Technological Innovation Programs of Higher Education
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