Abstract:Atherosclerosis (As) is a chronic inflammatory arterial wall injury process, and vessel wall cells play an important role in the occurrence and development of As. Vascular endothelial cell (VEC) act as a semi-permeable barrier between vascular smooth muscle cell (VSMC) and vascular lumen, and its injury is the initial stage of As. In addition, Through phenotypic transformation, VSMC could transform into many cell phenotype of the plaques, including macrophage, foam cell, mesenchymal stem cell and so on, and these cells further involved in the occurrence of As. Fibroblast is the main component of vascular adventitia,in pathological conditions, fibroblast differentiate into myofibroblast and participate in the occurrence of As. In this article, we will review the involvement of vascular wall cells in the mechanism of As and its potential therapeutic targets for the treatment of As, which provide new therapeutic ideas for As.

Abstract:Aim To investigate whether disturbed flow affects endothelial cell function and atherosclerotic plaque formation by regulating histone demethylase KDM5B and epigenetic modification. Methods After partial carotid artery ligation (PCL), single-cell data analysis and immunofluorescence staining were used to investigate the changes of histone methylation level and histone demethylase expression in carotid endothelial cells of wild type mice under perturbed flow. qPCR and Western blot were used to detect the expression of KDM5B and H3K4me3 in endothelial cells exposed to disturbed flow. Transcriptome sequencing was used to analyze the effect of KDM5B knockdown on endothelial cell function.Endothelial cell ring formation assay was used to verify the effect of KDM5B on angiogenesis. PCL combined with high-fat diet for 2 weeks was used to establish a carotid artery plaque model to analyze the effect of KDM5B knockdown on plaque formation. Results There was a large amount of H3K4me3 methylation in vascular endothelial cells. Blood disturbed flow reduced the methylation of H3K4me3 (P＜0.01) and promoted the expression of histone demethylases KDM5B in endothelial cells (P＜0.05). Compared with control group, inhibition of KDM5B activity or knockdown of KDM5B increased H3K4me3 level in endothelial cells (P＜0.05). Compared with Con313 control group, KDM5B knockdown reduced atherosclerotic plaque formation by 41.45% (Con313 control group:42.17%±1.90%, shKDM5B knockdown group:24.69%±1.60%, P＜0.01) by inhibiting angiogenesis. Conclusions Blood disturbed flow promotes KDM5B expression, reduces H3K4me3 modification, and promotes angiogenesis and atherosclerotic plaque formation. Targeting the KDM5B-H3K4me3 axis can be used as a candidate therapeutic target related to cardiovascular diseases.

Abstract:Endothelial dysfunction is a pivotal contributor to atherosclerosis (As) pathogenesis. A comprehensive understanding of the mechanisms of endothelial dysfunction would provide novel insights into effective treatment of As. Recent advances in genome and transcripome technology have enabled researchers to further explore the molecular mechanisms of endothelial dysfunction. It has been found that the regulatory network of competitive endogenous RNA (ceRNA) mediated by long non-coding RNA (lncRNA) plays a key role in endothelial dysfunction. lncRNA acts as a “molecular sponge” for microRNA (miRNA) to block the post-transcriptional repression of miRNA on downstream target gene messenger RNA (mRNA) by binding to miRNA, thereby regulating the function and phenotypic conversion of endothelial cell (EC) lncRNA-miRNA-mRNA interactions are widely involved in play an essential role EC inflammatory responses, apoptosis, autophagy, angiogenesis, and endothelial-mesenchymal transition (EndMT). Which suggests that it may be a potential therapeutic targets for As.

Abstract:Aim To explore the specific mechanism by which scutellarin (Scu) antagonizes the injury of human aortic endothelial cells (HAEC) induced by 2,2-azobis(2-methylpropylimidate) dihydrochloride (AAPH) by regulating the protein kinase RNA-like endoplasmic reticulum kinase (PERK)-nuclear factor erythroid 2-related factor 2 (NRF2)/activating transcription factor 4 (ATF4)-C/EBP homology protein (CHOP) pathway. Methods HAEC were pre-protected by Scu and then injured by AAPH to explore the molecular mechanism of Scu on HAEC injury. The cells were divided into control group, AAPH group, AAPH＋Scu low, medium and high groups. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and glutathione S-transferase (GSH-ST) in the cells were measured. The content of reactive oxygen species (ROS) in cells was detected by fluorescent probe, and the apoptosis rate was detected by Annexin V-FITC/PI method. The mRNA expression of PERK and eIF2α in cells was detected by RT-qPCR. The protein expression of glucose regulated protein 78 (GRP78), PERK, p-PERK, eukaryotic initiation factor 2α (eIF2α), p-eIF2α, ATF4, CHOP, Nrf2, Bcl-2, p53 up-regulated modulator of apoptosis (PUMA), Caspase-3 and Caspase-12 in cells was detected by Western blot. In order to further study the molecular mechanism of Scu against HAEC injury, gene silencing technology was used to inhibit the expression of PERK in HAEC. The cells were divided into five groups:control group, AAPH＋si-con group, AAPH+Scu+si-con group, AAPH+si-PERK group, AAPH+si-PERK+Scu group. The mRNA expression of PERK and eIF2α in cells after si-PERK interference was detected by RT-qPCR. The protein expression of PERK, p-eIF2α, eIF2α, ATF4, CHOP, Nrf2, Bcl-2, PUMA, Caspase-3 and Caspase-12 in cells after si-PERK interference was detected by Western blot. Results The content of ROS and the rate of apoptosis were significantly reduced after Scu intervention (P＜0.01). Scu could down-regulate the mRNA expression of PERK and eIF2α, and down-regulate the protein expression of GRP78, p-PERK, p-eIF2α, ATF4, CHOP, PUMA, Caspase-3, Caspase-12 and up-regulate the protein expression of Nrf2 and Bcl-2 (P＜0.01). After interference with si-PERK, there were significant differences in the protein expression of PERK, p-eIF2α, ATF4, CHOP, Nrf2, Bcl-2, PUMA, Caspase-3, Caspase-12, as well as the mRNA expression of PERK and eIF2α in cells compared to before interference (P＜0.01). It is proved that Scu could anti-endoplasmic role in reticulum stress and apoptosis, which is closely associated with the regulation of the PERK-Nrf2/ATF4-CHOP pathway. Conclusion Scu can effectively alleviate AAPH-induced injury to HAEC by regulating PERK-Nrf2/ATF4-CHOP pathways to inhibit endoplasmic reticulum stress and cell apoptosis.

Abstract:Atherosclerosis (As) is one of the main causes of death in the global population. The main causes are hyperlipidemia, hypertension, obesity, etc. However, after avoiding traditional risks, As may still progress. In recent years, the role of microbial infection factors in As has been increasingly valued. This article reviews the correlation between infection and As, and focuses on introducing the research progress on the mechanisms by which microbial infections such as Helicobacter pylori, Chlamydia pneumoniae, human immunodeficiency virus (HIV), and cytomegalovirus promote the development of As, aiming to discover the correlation between microbial infections and As and provide new ideas for the prevention and treatment of As.

Abstract:Aim Biomarkers of vascular endothelial injury serve as crucial indicators for assessing vascular endothelial function and provide a basis for the diagnosis and treatment of cardiovascular diseases (CVD). Currently, the mechanisms and clinical significance of classical biomarkers, such as C-reactive protein and intercellular adhesion molecule, in CVD have been extensively studied. However, their limited sensitivity and specificity make it difficult to comprehensively reflect the progression of CVD. With the advancement of liquid biopsy and novel molecular biology detection technologies, emerging biomarkers of vascular endothelial injury have become a research focus. This article systematically reviews biomarkers of vascular endothelial injury, with a particular emphasis on the research progress and clinical applications of novel biomarkers in CVD.

Abstract:Macrophages play multiple roles in atherosclerosis. The progression of atherosclerosis is associated with morphological and functional changes in the lymphatic vessels of the diseased arteries, but the mechanism is not fully understood. This paper mainly reviews the origin and classification, the markers and the function of macrophage in atherosclerosis, the origin, the structure function and the markers of lymphatic vessels in atherosclerosis, the changes of arterial wall lymphangiogenesis in different stages of atherosclerotic lesions, the functional role of lymphangiogenesis in atherosclerosis, the lymphatic migration of macrophages and its mechanism involved in lymphangiogenesis, in order to provide a basis for the mechanism research and clinical treatment of atherosclerosis.

Abstract:Aim Krüppel-like factor (KLF) 2 and 4 are two core transcription factors closely related to vascular homeostasis, with multiple protective effects such as anti-inflammatory, anti-calcification and anti-thrombotic. The aim of this study is to elucidate and validate the vascular homeostasis related gene profile co-regulated by KLF2 and KLF4 in endothelial cells. Methods Human umbilical vein endothelial cells (HUVEC) were treated with adenovirus (Ad-KLF2 or Ad-KLF4) and control virus (Ad-NC) for 24 h, RNA was extracted from the cells and analyzed by transcriptomic sequencing. The sequencing results of overexpressed KLF2 and KLF4 were superimposed with the sequencing results of reported KLF2/KLF4 double-gene knockout mice. The selected differential expression genes were verified by real-time fluorescence quantitative PCR in HUVEC treated with Ad-KLF2 or Ad-KLF4, and in HUVEC treated with atorvastatin or resveratrol. Results Transcriptomic superposition revealed 256 differential expression genes were up-regulated by KLF2 and KLF4, and KEGG analysis showed that differential expression genes were enriched in hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, dilated cardiomyopathy, ECM-receptor interaction and focal adhesion; there were 145 differential expression genes down-regulated by KLF2 and KLF4, and KEGG analysis showed that differential expression genes were enriched in microRNA of cancer, mineral absorption, glycosaminoglycan biosynthesis-chondroitin sulfate/dermatan sulfate, p53 signaling pathway and biosynthesis of amino acids. Finally, six novel genes regulated by KLF2 and KLF4 were obtained. Conclusion FGFR3, SEMA4B, SEMA6A, PTX3, FABP4 and FABP5 may be novel genes that regulate vascular homeostasis in endothelial cells by the transcription factors KLF2 and KLF4.

Abstract:Aim To explore the mechanism of oxidized lipoprotein(a) (oxLp(a) inducing pyroptosis of vascular endothelial cells. Methods After incubating human umbilical vein endothelial cells (HUVEC) with 100 mg/L oxLp(a) for 24 hours, Western blot and RT-qPCR was used to detect pyroptosis related proteins, pro-inflammatory cytokines, mitochondrial related proteins NRF1, NRF2, PGC-1α and mitochondrial gene cytochrome b (CYTB), ELISA was used to detect the levels of inflammatory factors, scanning electron microscopy was used to detect cell membrane rupture, transmission electron microscopy was used to detect mitochondrial morphology, Hoechst33342/PI staining was used to detect cell apoptosis, MitoSOX probe was used to detect mitochondrial reactive oxygen species (mtROS), Flu-4AM probe was used to detect calcium ions, JC-1 probe was used to detect mitochondrial membrane potential (MMP), and Calcein AM staining was used to detect mitochondrial permeability transition pore (mPTP). Transfecting HUVEC with CYTB overexpressing lentivirus and analyzing its effects on oxLp(a) induced pyroptosis and mitochondrial function. ResultsAfter treatment with oxLp(a), the expression of NLRP3, pro-Caspase-1, Caspase-1, GSDMD and GSDMD-N proteins related to pyroptosis were significantly increased (P＜0.05); the protein and mRNA levels of CYTB and pro-inflammatory cytokine IL-1β, IL-18 were significantly increased (P＜0.05). Small pores appeared on the cell membrane, the percentage of PI stained positive cells significantly increased (P＜0.05). OxLp(a) significantly inhibited the expression of mitochondrial related proteins NRF1, NRF2 and PGC-1α, and the expression of mitochondrial gene CYTB, promoted an increase in mtROS generation, Ca2＋ overload, a decrease in ATP levels, a decrease in MMP, an increase in mPTP values, and abnormal mitochondrial morphology. After transfection with pHelper 2.0 lentivirus vector overexpressing CYTB, it was found that oxLp(a) induced HUVEC pyroptosis and mitochondrial morphological and functional abnormalities were partially reversed by overexpression of CYTB. Conclusion oxLp(a) promotes mitochondrial morphological and functional abnormalities and induces HUVEC pyroptosis by downregulating CYTB.

Abstract:Atherosclerosis (As) is the pathological basis of various vascular diseases, which affects the function of important organs. Recent studies have found that N6-methyladenosine (m6A) modification plays a vital role in As. This paper summarizes the regulatory effects of m6A modification on vascular endothelial cell (VEC) injury, vascular smooth muscle cell (VSMC) transformation, macrophage (M) differentiation, foam cell (FC) formation, pyroptosis, and lipid regulation, and summarizes the regulation function of some m6A regulatory proteins in As.