Abstract:Aim To study the regulatory effect of acupuncture pretreatment on ferroptosis of nerve cells in rats with ischemic stroke. Methods Sixty SD rats were randomly divided into sham middle cerebral artery occlusion (MCAO) group, MCAO group, and acupuncture+MCAO group. In the acupuncture+MCAO group, the acupuncture points of DU26, PC6, and SP6, were selected for acupuncture pretreatment, once a day for a total of 5 days. After pretreatment, MCAO or sham MCAO models were prepared. The Zausinger sextintegral method was used to score the neurological function of rats, and the infarct volume of brain tissue was calculated by TTC staining. Electron microscopy was used to observe the pathomorphological changes of brain tissue. The iron content was detected by colorimetric method, the content of malondialdehyde (MDA) and glutathione (GSH) was determined by ELISA, and the expression of glutathione peroxidase 4 (GPX4) was detected by immunofluorescence. Results Compared with the sham MCAO group, the MCAO group had a decrease in neurological function scores, a significant increase in infarct volume, a decrease in the number of mitochondria under electron microscopy, a rupture and vacuolization of the inner mitochondrial cristae, an increase in the contents of iron and MDA in brain tissue, and a decrease in GSH content and GPX4 expression. Compared with the MCAO group, the acupuncture+MCAO group had an increase in neurological function scores, a decrease in infarct volume, a large number of mitochondria under electron microscopy, a clear structure, a decrease or disordered arrangement of some mitochondrial crest structures, a decrease in the contents of iron and MDA in brain tissue, and an increase in GSH content and GPX4 expression. Conclusion Acupuncture pretreatment can alleviate neurological damage in rats, and its mechanism may be related to regulating iron, GSH and MDA contents in brain tissue, and GPX4 expression, improving cell antioxidant capacity and inhibiting nerve cell ferroptosis.

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:At present, with the development of new diagnostic technologies, the detection rate of aortic dissection has been increasing year by year, but its mortality rate still remains high. Cardiovascular disease is a chronic inflammatory disease, and vascular inflammation plays a major role in the progression of aortic dissection. Therefore, this article systematically describes the specific roles and mechanisms of inflammatory cells, inflammatory factors, and inflammasomes in the development of aortic dissection.

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 impact and mechanism of proprotein convertase subtilisin kexin 9 (PCSK9) on the progression of abdominal aortic aneurysm (AAA). Methods 6～8 week old ApoE－/－ mice were selected to establish the AAA model. AngiotensinⅡ (AngⅡ) was continuously infused through subcutaneous implantation of a micro-osmotic pump. The mice were fed with high-fat diet and killed after 28 days. The expression of PCSK9 in abdominal aortic smooth muscle cells was detected by immunohistochemistry and immunofluorescence in normal abdominal aortic blood vessels and AAA samples in human and mice. Primary cultured murine vascular smooth muscle cells (mVSMC) of C57BL/6 mice were treated with different concentrations of AngⅡ for 24 h, and the expression of PCSK9 mRNA and protein was detected. PCSK9 overexpression and knockdown cell models were established, and mitochondrial reactive oxygen species (mtROS), mitochondrial membrane potential (MMP), mitochondrial permeability transition pore (MPTP) opening, and Z-DNA binding protein 1 (ZBP1) protein expression were detected. Bioinformatics was used to analyze the differential expression of multiple single-cell sequencing datasets to obtain the key differentially expressed genes, and to study their expression and role in AAA. Results Immunohistochemistry and immunofluorescence results showed that PCSK9 expression in human and mouse AAA increased (P＜0.01), and co-localized with smooth muscle. AngⅡpromoted PCSK9 expression in mVSMC in a concentration-dependent manner, the 2.0 μmol/L AngⅡ group showed a 2.9-fold and 1.1-fold increase in the expression of PCSK9 mRNA and protein, respectively (P＜0.01), with the most significant effect observed. After successfully constructing PCSK9 overexpression and PCSK9 interference mVSMC models, PCSK9 overexpression led to an increase in intracellular mtROS, a decrease in MMP, an increase in MPTP opening, and a decrease in cellular activity (P＜0.01); PCSK9 knockdown could reduce AngⅡinduced increase in mtROS, decrease in MMP and MPTP opening; compared with the siNC＋AngⅡgroup, the siPCSK9＋AngⅡ group showed a decrease in mtROS and an increase in the fluorescence brightness of MMP and MPTP (P＜0.05). Bioinformatics analysis revealed that ZBP1 was a core differentially expressed gene in AAA. Immunohistochemistry and immunofluorescence results showed that ZBP1 expression in human and mouse AAA tissues increased, and co-localized with smooth muscle. Western blot results showed that PCSK9 overexpression or treatment with 2.0 μmol/L AngⅡ could increase ZBP1 protein expression (P＜0.01), while PCSK9 knockdown could alleviate the increased ZBP1 expression caused by AngⅡ (P＜0.05). Conclusion PCSK9 may induce mitochondrial damage in smooth muscle cells, activate downstream molecule ZBP1 to cause cell damage, and promote the development of AAA.

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 is a chronic vascular disease, whose early manifestations are vascular intimal hyperplasia and plaque formation, and the late vascular events can be caused by plaque rupture after erosion. In the process of atherosclerosis, vascular smooth muscle cell(VSMC) in the arterial media play a key role in vascular remodeling, and their phenotype conversion has an important impact on maintaining vascular homeostasis. Due to methodological limitations, the previous understanding of VSMC function is limited; With the rapid development of lineage tracing and single-cell sequencing technology, the role of VSMC in atherosclerosis has been deeply explored in recent years. This paper summarizes the emerging research methods of VSMC in recent years, and reviews the literature on the roles and mechanisms of phenotypic switching of VSMC in vascular remodeling.

Abstract:Aim Ischemic stroke (IS) is caused by acute ischemia of cerebral blood vessels, leading to brain tissue damage and neuronal apoptosis. The pathogenesis is complex, involving multiple cell death modes such as pyroptosis, ferroptosis and disulfide death. Disulfide death is a newly discovered form of death that helps to explore the pathological mechanisms of various diseases from a new perspective. The aim of this study is to discover and validate the differential expression of disulfide death-related genes in blood samples of ischemic patients and their association with immune regulation. Methods The relevant datasets of clinical patients (GSE16561 and GSE37587) were obtained through online big data. Differentially expressed genes related to disulfide death were identified, and gene enrichment analysis was conducted to further explore the potential mechanisms. Subsequently, immune cell infiltration was analyzed to investigate the dysregulation of immune cells in the context of IS. Finally, the accuracy of key genes was verified through ROC curves, column charts, calibration curves, and decision curves, and a disease prediction model was constructed to predict the risk of stroke. Results Based on this dataset, significant differential expression of 9 genes related to disulfide death was identified. Independent external validation was conducted using the microarray dataset GSE58294. Single item comparisons were performed on these differentially expressed genes in blood samples from 69 IS patients and 23 normal individuals.The results showed that the trends of LRPPRC, MYH9, NDUFA11, PRDX1 and RPN1, the 5 differentially expressed genes, were consistent. Immune infiltration analysis found that differentially expressed genes such as TLN1, MYH9, PRDX1, LRPPRC, NDUFA11 were also strongly correlated with CD8＋T cells, activated NK cells, macrophages, and neutrophils in IS patients. Functional enrichment analysis emphasized the important role of pathways such as focal adhesion, platelet aggregation, and activation in the occurrence and development of diseases. By using a column chart model for risk prediction, it was shown that the accuracy of these differentially expressed genes was good, and the ROC curve AUC value of the optimized combination of disulfide death-related genes could reach 0.844. Further validation through an external dataset (GSE58294) revealed that the ROC curve AUC value optimized for disulfide death-related genes reached 0.989, which had good clinical guidance significance for the risk of IS. Conclusions This study confirmed the existence of 5 disulfide death-related genes in IS patients through a dataset, including upregulation of MYH9 and downregulation of LRPPRC, NDUFA11, PRDX1 and RPN1. These gene alterations are suggested to influence IS disease progression and prognosis through immune inflammation and bleeding risk.

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.