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:Endothelial dysfunction, a common feature of various cardiovascular diseases, is closely associated with the overexpression of reactive oxygen species (ROS)/reactive nitrogen species (RNS). The reaction between superoxide anion and nitric oxide (NO) can generate peroxynitrite with stronger oxidation ability, which can deplete NO by oxidizing various proteins, leading to endothelial contraction and relaxation dysfunction, and playing an important role in various cardiovascular diseases. This article reviews the pathways through which nitrosylation modified proteins are produced and the possible mechanisms by which they promote endothelial dysfunction in cardiovascular disease. It discusses the relationship between ROS/RNS mediated nitrosylation modification and endothelial dysfunction, which together promote the progression of cardiovascular disease. The article also discusses the application of therapeutic strategies such as clearing peroxynitrite, inhibiting ROS production pathways, and directly enhancing endothelial cell function in cardiovascular diseases related to endothelial dysfunction, which can provide reference for further research on the role of protein nitration modification as a post-translational intervention target in cardiovascular diseases.

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 To investigate the effect of CDR132L (miR-132 antisense oligonucleotide) on vascular remodeling and function in mice with hypertension and hyperlipidemia, and explore its possible mechanism. Methods A total of 30 8-week-old male C57BL/6 mice were randomly divided into three groups:control group, model group and CDR132L group, with 10 mice in each group. The control group received with a standard diet while the model group and CDR132L group received N-nitro-L-arginine methyl ester (L-NAME) and high-fat diet to induce hypertension and hyperlipidemia. The CDR132L group was administered with intraperitoneal injection of CDR132L at a dose of 20 mg/kg once weekly for six consecutive weeks, whereas the control group and the model group were given intraperitoneal injection of an equivalent volume of normal saline. The tail-cuff method was utilized for blood pressure measurement, blood lipid and glucose levels were assayed by an automatic biochemical analyzer, the thoracic aorta structure was observed by HE staining, endothelium-dependent relaxation of the thoracic aorta was evaluated by the vascular ring test, the expression level of miR-132 in the thoracic aorta was measured by qPCR, the protein expression levels of Gab1 and endothelial nitric oxide synthase (eNOS) in the thoracic aorta were determined by Western blot. Results Compared with the control group, the model group demonstrated notable rises in systolic and diastolic blood pressure, serum triglyceride, total cholesterol levels, and body weight. Moreover, the intima of thoracic aorta and the thickness of vascular wall was uneven, the smooth muscle cells of the tunica media were arranged irregularly, with a large amount of fat deposition in the vascular wall, and the endothelium-dependent relaxation response of thoracic aorta was decreased (P＜0.05). The expression level of miR-132 in the thoracic aorta was significantly increased (P＜0.05), while the expression level of Gab1 and eNOS protein was markedly decreased (P＜0.05). Compared with the model group, the CDR132L group showed no significant differences in systolic and diastolic blood pressure, serum triglyceride and total cholesterol levels, as well as body weight (P＞0.05).However, the CDR132L group exhibited a complete and smooth intima of the thoracic aorta with minimal intravascular lipid deposition, the thickness of the vascular wall was uniform, the smooth muscle cells of the tunica media were arranged orderly, accompanied by enhanced endothelium-dependent relaxation response of the thoracic aorta (P＜0.05). The expression level of miR-132 in the thoracic aorta was significantly decreased (P＜0.05), while the expression levels of Gab1 and eNOS protein were significantly increased (P＜0.05). Conclusion CDR132L can improve vascular remodeling and endothelium-dependent relaxation in hypertensive and hyperlipidemia mice, which may be related to the decrease of miR-132 expression level and the up-regulation of Gab1 and eNOS protein expression levels in the thoracic aorta.

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.