Patients with systemic lupus erythematosus (SLE) have significantly increased cardiovascular risk, and atherosclerosis (AS) is a major cause of long-term mortality. However, whether SLE-specific immune dysregulation drives accelerated AS remains unclear. This study explored the role of interferon-stimulated gene 15 (ISG15) in SLE-associated AS. We first characterized the disease manifestations, then identified disease characteristics at the proteomic and transcriptomic levels via multi-omics analysis of a pristane-induced SLE-ApoE−/− mouse model and integrated analysis of SLE and AS datasets. We found ISG15 was uniquely elevated across atherosclerotic plaque types and correlated with cardiovascular events, with robust interferon-alpha response activation in SLE mice. Further, ISG15 colocalized with CD68⁺ macrophages in plaques, and extended analysis showed it was highly expressed in peripheral blood monocytes of SLE patients. Clinically, ISG15⁺ monocyte proportion was significantly higher in SLE patients with AS, outperforming conventional cardiovascular risk scores and serving as an independent predictor. In conclusion, ISG15 is a critical effector linking SLE immune dysregulation to accelerated AS, and ISG15⁺ monocyte proportion is a promising biomarker for cardiovascular risk stratification in SLE patients.

Flow cytometry, as a groundbreaking technique in single-cell analysis, has evolved into a multi-dimensional technical framework over six decades of development. This paper systematically reviews its technological progression, ranging from conventional fluorescence labeling to spectral unmixing in spectral flow cytometry, advancements in metal tagging within mass cytometry, morphological and functional synchronous analysis in imaging flow cytometry, and ultimately achieving sub-micron-level detection in nanoparticle flow cytometry. Each technological advancement has expanded the boundaries of biological understanding. These innovations have substantially enhanced the sensitivity and dimensionality of single-cell analysis, and have thereby facilitated a range of clinical applications, which include tumor immunotherapy monitoring, cardiovascular disease risk assessment, and precise diagnosis of infectious diseases. Nevertheless, challenges remain in high-dimensional data processing, standardization protocols, and the translation of emerging technologies. This article aims to elucidate the technical context, highlight its impact on precision medicine, and provide a reference framework for interdisciplinary research.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, and its course is often accompanied by multiple organ damage. The mortality rate of SLE exhibits a "bimodal pattern", namely the early death peak is primarily attributed to infection and lupus activity, while the late death peak lists cardiovascular diseases (CVD) caused by atherosclerosis (AS) as the leading cause of death. Mitochondria, as the hub of energy metabolism and the multi-dimensional regulatory center of cellular functions, play a key role in the occurrence and development of AS plaques under the pathological background of SLE. This review systematically sorted out the mitochondrial dysfunction mechanisms of different immune cells and endothelial cells in SLE, and deeply expounded their influence pathways on the pathological process of AS. Furthermore, this article explores the current clinical treatment strategies for SLE and analyzes the therapeutic potential of mitochondrial-targeted intervention measures.

Atherosclerosis (AS)，a major global cause of cardiovascular mortality，is characterized by significant metabolic reprogramming of plaque macrophages in response to a hostile microenvironment containing cytokines, oxidized lipids, and hypoxia. Macrophage metabolic reprogramming, marked by shifts in glycolysis, fatty acid, and amino acid metabolism has emerged as a critical contributor to chronic inflammatory diseases. This study explores the role of fructose-1,6-bisphosphatase 1 (FBP1) in this process and its functional interplay with hypoxia-inducible factor-1α (HIF-1α). Clinically, FBP1 and HIF-1α levels were significantly elevated in peripheral blood mononuclear cells (PBMCs) and serum from AS patients. ROC analysis indicated their strong potential as diagnostic biomarkers. These findings were corroborated in high-fat diet-fed mice and ox-LDL-stimulated macrophages, which showed increased FBP1 and HIF-1α expression in atherosclerotic lesions and immune cells. FBP1 overexpression mitigated ox-LDL-induced metabolic reprogramming, evidenced by reduced lactate production, reactive oxygen species (ROS) generation, and lipid droplet accumulation. Conversely, FBP1 suppression exacerbated these metabolic alterations. Mechanistically, FBP1 directly interacted with and inhibited the expression of HIF-1α. Inhibition of HIF-1α reverses the exacerbation of cellular metabolic reprogramming induced by FBP1 inhibition. In summary, the results demonstrate FBP1/ HIF-1α axis as mediator of macrophage metabolic reprogramming in AS, highlighting its dual significance as a contributor to disease pathogenesis and a promising basis for clinical diagnosis.

BACKGROUND:Atherosclerosis is a primary contributor to worldwide morbidity and mortality. Failure to timely clear apoptotic cells can trigger a cascade reaction, where the necrotic core expands until the fibrous cap is ruptured, and atherosclerotic plaques become vulnerable. Efferocytosis is an important method for recognizing and eliminating apoptotic cells. Nevertheless, the specific effect of efferocytosis on atherosclerosis remains uncertain. This study aimed to identify and verify the relevant characteristics of efferocytosis for detecting atherosclerosis. METHODS:The data of gene expression patterns of atherosclerosis were sourced from the Gene Expression Omnibus (GEO) database, and the differential expression analyses of efferocytosis-related genes (EFRGs) were performed between the atherosclerosis samples and the control samples. Subsequently, protein-protein interaction (PPI), correlation analysis, and functional enrichment analysis were performed to reveal the interaction between molecules as well as their pathways. Machine learning (ML) was employed to determine hub genes to construct a clinical prediction model. At the same time, immune infiltration, single-cell transcriptome analysis, and cell experiments were conducted in both atherosclerosis and control samples to provide a reference for the immune cell landscape and the cell heterogeneity under this condition. RESULTS:The study revealed that 14 genes were closely related to efferocytosis in atherosclerosis. Among them, an ML model was used to screen 5 potential diagnostic biomarkers, including tumor necrosis factor (TNF), apolipoprotein E (ApoE), neutrophil cytosolic factor 1 (NCF1), triggering receptor expressed on myeloid cells 2 (TREM2), and chitinase-3 like-protein-1 (CHI3L1). Subsequent external validation indicated that, except for TNF, the other 4 genes were all upregulated. From the cell-type identification by estimating relative subsets of RNA transcripts (CIBERSORT) analysis, those 5 genes were all significantly associated with various immune cells. Further single-cell RNA sequencing (scRNA-seq) analysis demonstrated that those 5 genes were selectively upregulated in the macrophages of atherosclerosis lesions, which was supported by mRNA levels in cell experiments. CONCLUSIONS:This study clarified the association between atherosclerosis and efferocytosis, and established an effective diagnostic model. Moreover, potential treatment targets for atherosclerosis were identified, offering new insights into the potential mechanism of atherosclerosis.

Background: Autophagy exerts a vital role in the development of atherosclerotic lesions. Mounting evidence suggests a significant link between autophagy and atherosclerosis. Methods: Two atherosclerotic plaque datasets were integrated from the Gene Expression Omnibus (GEO) database. After differentially expressed genes (DEGs) were determined, enrichment analyses were subsequently performed on DEGs. We employed weighted gene coexpression network analysis (WGCNA) and cross-linked these modules with DEGs and autophagy-related genes. Subsequently, a prediction model was established for evaluation. RT-PCR was adopted to identify hub gene expression. The consensus clustering analysis on the overlapping genes was executed. Evaluation of immune infiltration was conducted on the merged dataset. A TF-miRNA-mRNA regulatory network was then established for the hub genes. Results: The differential gene expression analysis uncovered 259 DEGs. Enrichment analysis showed that immune and inflammatory reactions were related to atherosclerosis. By intersecting DEGs, WGCNA module genes, and ARGs, 13 overlapping genes were obtained. Four machine learning models identified seven hub genes. Furthermore, six of the seven genes demonstrated potential for disease diagnosis. The prediction model, based on the expression levels of these six genes, yielded satisfactory results. RT-PCR analysis demonstrated that the mRNA expression of six genes meets expectations. Consensus clustering divides 13 overlapping genes into two clusters, C1 and C2, with significant differences in immune infiltration. Immune cell infiltration demonstrated heightened immune activity within the atherosclerotic plaque group. A TF-miRNA-mRNA regulatory network was established for the six genes. Conclusion: It is anticipated that these six genes may serve as significant and valuable targets for future research into atherosclerosis.

Systemic lupus erythematosus (SLE) is an autoimmune disease that significantly increases the risk of cardiovascular diseases, particularly atherosclerosis (AS). Understanding the shared pathogenic mechanisms underlying SLE and AS is crucial for developing effective therapeutic strategies. Macrophages, as pivotal immune cells, play a critical role in the initiation and progression of atherosclerotic plaques within the context of SLE. This review delves into the molecular and cellular mechanisms governing macrophage activation and differentiation in response to SLE-related inflammatory mediators, highlighting their roles in lipid metabolism, plaque stability, and immune regulation. Additionally, we discussed the current treatment modalities for SLE and their impact on macrophage functionality, exploring these effects for atherosclerotic progression. By elucidating the intricate relationship between macrophages, SLE pathophysiology, and AS progression, this review underscores the need for a multidisciplinary approach in managing SLE and its cardiovascular complications, aiming to improve patient survival and quality of life through tailored therapeutic interventions addressing both autoimmune and cardiovascular pathologies.

ObjectiveAtherosclerosis (AS) is the underlying pathology of atherosclerotic cardiovascular disease and a major cause of cardiovascular-related mortality. Chronic inflammation and mitochondrial dysfunction, triggered by the infiltration of various immune cells, are key factors in the progression of AS. However, the interaction and crosstalk between these factors remain unclear.MethodsTwo gene expression datasets, GSE100927 and GSE43292, were downloaded from the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) database, covering carotid atherosclerosis and control groups. After identifying the common differentially expressed genes (DEGs), mitochondria-related DEGs (Mito-DEGs) were obtained through Weighted Gene Co-expression Network Analysis (WGCNA) and machine learning approaches. Immune infiltration analysis and comparison were subsequently performed. The single-cell dataset GSE159677 further validated the expression proportions of relevant genes in different cell populations during the progression of AS. Additionally, cell-cell communication and trajectory analysis within the immune landscape were utilized to infer the pathways of cell state transitions within AS clusters. THP-1 cells were cultured in vitro, and the foam cell model was established by the addition of oxidized low-density lipoprotein (ox-LDL). The expression trends of hub Mito-DEGs were confirmed via qRT-PCR.ResultsFrom the GSE100927 and GSE43292 datasets and the MitoCarta3.0 database, three hub Mito-DEGs closely associated with AS were ultimately identified: CASP8, GATM, and LAP3. Subsequent comprehensive bioinformatics analysis of these hub genes further emphasized the importance of the immune system in AS. Immune profiling based on the CIBERSORT algorithm revealed significantly increased infiltration of activated mast cells, monocytes, memory B cells, T follicular helper cells, and M0 macrophages in the immune microenvironment of AS. Single-cell analysis showed that GATM and LAP3 were enriched in monocytes and macrophages, while CASP8 exhibited increased expression in NK cells, T cells, and monocytes. In addition, in vitro cell experiments demonstrated that mRNA expression levels of the hub Mito-DEGs were significantly elevated in the lipid-laden foam cell group compared to the control group, consistent with the expression patterns observed in the single-cell dataset.ConclusionThis study revealed the interaction between Mito-DEGs and the immune system in AS. These findings may provide new insights into therapeutic monitoring and prognosis evaluation.

Background: Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1)'s role has been shown to drive immune regulation and inflammation in many human diseases. However, the exact mechanism of action of IFIT1 in AS is unclear, and the specific mechanism of action on METs is also unknown. In this study, we will explore the potential mechanisms of IFIT1 in the formation of METs during AS. Methods: We downloaded GSE100927, GSE193336, GSE159677, IRGs, and METs-related genes for analysis and used qRT-PCR, flow cytometry, and immunofluorescence to detect the expression levels of IFIT1 and METs in plaques from AS patients and mice. The potential association of IFIT1 and METs in macrophages was similarly verified in LPS-induced macrophages. After IFIT1 silencing, the expression levels of METs were detected using qRT-PCR, flow cytometry, immunofluorescence, and WB. In addition, we delved into the potential mechanisms to detect the expression of the STING-TBK1 pathway and explored the interaction between IFIT1 and the STING-TBK1 pathway. Results: Our results showed that IFIT1 was upregulated in AS patients, mouse plaque tissues, and LPS-induced macrophages. The same changes were observed in METs.The decrease in METs after IFIT1 silencing suggests that IFIT1 is involved in the regulation of macrophages through METs. Notably, with the decrease in IFIT1 levels, we observed a corresponding decrease in the STING-TBK1 pathway, which decreased accordingly, suggesting some connection between IFIT1, STING-TBK1, and METs. Validation of the effect of STING-TBK1 on a macrophage basis showed that the STING activator SR-717 increased the expression of METs, while the STING inhibitor H-151 had the opposite result. Interestingly, we added SR-717 and H-151 to si-IFIT1, respectively, and the same changes occurred in METs. Conclusion: In summary, our study suggests that IFIT1 activates METs through the STING-TBK1 pathway, thereby aggravating AS.

Atherosclerosis (AS), the leading cause of cardiovascular diseases, is heavily influenced by inflammation, lipid accumulation, autophagy, and aging. The expression of glycoprotein non-metastatic melanoma B (GPNMB) has been observed to correlate with lipid content, inflammation, and aging, progressively increasing as atherosclerosis advances through its various stages, from baseline to early and advanced phases. However, the interaction between GPNMB and AS is controversial. Knockout of GPNMB has been shown to increase atherosclerotic plaque burden in mice. Conversely, targeted elimination of GPNMB-positive cells reduced atherosclerotic burden. These seemingly contradictory findings underscore the complexity of the issue and highlight the need for further research to reconcile these discrepancies and to elucidate the precise role of GPNMB in the pathogenesis of AS.

Tumor-associated autoantibodies (TAAS), as cancer biomarkers, have attracted special attention. In recent years, increasing evidence has indicated that TAAS shows an elevated level in the early stage of human malignancies, and examination of TAAS in patients′ clinical specimens has a good predictive value for a variety of cancers′ early diagnosis. The mechanism of TAAS and its clinical application will be introduced, and the advantages and problems of tumor autoantibodies as markers will be expounded in this article.

Recently, trimethylamine N-oxide (TMAO) has been considered a risk factor for cardiovascular disease and has a proatherogenic effect. Many studies have found that TMAO is involved in plaque oxidative stress and lipid metabolism, but the specific mechanism is still unclear. In our study, meta-analysis and bioinformatic analysis were firstly conducted in the database, and found that the effect of high plasma TMAO levels on promoting atherosclerotic plaque may be related to the expression of key antioxidant genes nuclear factor erytheroid-derived-2-like 2 (NFE2L2/Nrf2) decreased. Next, we assessed the role of Nrf2-mediated signaling pathway in TMAO-treated foam cells. Our results showed that TMAO can inhibit the expression of Nrf2 and its downstream antioxidant response element such as heme oxygenase-1 (HO-1) and glutathione peroxidase4 (GPX4), resulting in increased production of reactive oxygen species and decreased activity of superoxide dismutase, promoting oxidative stress. And TMAO can also promote lipid accumulation in foam cells by inhibiting cholesterol efflux protein expression. In addition, upregulation of Nrf2 expression partially rescues TMAO-induced oxidative stress and reduces ATP-binding cassette A1 (ABCA1)–mediated lipid accumulation. Therefore, TMAO promotes oxidative stress and lipid accumulation in macrophage foam cells through the Nrf2/ABCA1 pathway, which may provide a potential mechanism for the proatherogenic effect of TMAO.

Atherosclerosis is a chronic inflammatory response with lipid metabolism disorder.NOD-like receptor protein 3(NLRP3)inflammasome,as a multi-protein inflammatory complex,is closely related to cell activity,vascular inflammatory,and the progression of plaque.Trimethylamine oxide,as a major metabolite of intestinal flora,can initiate the activation of NLRP3 inflammasome and participate in the pathophysiological mechanism of atherosclerotic plaque formation and plaque rupture.The paper reviews the role of NLRP3 inflammasome and trimethylamine oxide in atherosclerosis to provide a new perspective for the mechanism research and clinical prevention and treatment of atherosclerosis.

Abstract Macrophages inflammation from variety of risk factors is critical in the rupture of atherosclerotic(AS) plaques. Trimethylamine oxide (TMAO), a dietary metabolite that depends on Gut microbiota, exerts strongly pro-inflammatory effects on Atherosclerosis. Nowadays, mounting research showed that NLRP3 inflammasome activation is essential for the pathogenesis of AS. The present study was to investigate the effect of TMAO on ox-LDL-induced NLRP3 inflammasomes activation of THP-1 cells, and to explore the potential mechanism. Used Cell Counting Kit-8 assay, LDH assay to evaluate the changes of macrophage activity under TMAO and ox-LDL, respectively, to clarify the appropriate dosage. Proteins related to NLRP3 inflammasomes, NF-kb and ERS were determined via Western Blot. Inflammatory cytokine secretion was then examined via ELISA. PCR detected gene levels of inflammatory markers, and caspase-1 activity assay was utilized to detect intracellular caspase-1 activity. The results showed that TMAO could activate NLRP3 inflammasome in ox-LDL-induced THP-1 macrophages and accelerate inflammatory factor release. In addition, TMAO can further activate the expression of ERS-related proteins( including BiP、p-PERK) and NF-KB pathway. NLRP3 inhibitor MCC950, NF-kb inhibitor JSH-23, and ER stress inhibitor 4-PBA reversed TMAO's promoting effect in ox-LDL-induced macrophage. Given these data, we conclude that TMAO promotes NLRP3 inflammasomes activation via the ERS/NF-kb pathway in ox-LDL induced THP-1 macrophages. Reducing the TMAO levels may be a viable approach to prevent atherosclerosis plaque development.

The progression of atherosclerotic plaque is accelerated by death of foam cells during the development of the plaque. There are several forms of foam cell death, such as autophagy and ferroptosis forms of cell death together are commonly predominant. Therefore, it is particularly important to study the crosstalk between various forms of cell death in atheroscler and ferroptosis. Although there is a dominant form of cell death that plays a role in the disease, motic plaques. Nuclear factor NF-E2-related factor (Nrf2) has been considered as a major regulator of antioxidant in previous studies, but recent studies have revealed that insufficient cellular autophagy can turn off Nrf2-mediated antioxidant defense while initiating Nrf2-manipulated iron deposition and lipid peroxidation, leading to the development of iron ferroptosis. The present experiment aimed to explain the regulatory mechanism between autophagy and ferroptosis through Nrf2. In this experiment, differentiated human THP-1 macrophages were used, which were treated with ox-LDL into foam cells with the addition of the autophagy inhibitor chloroquine (CQ), the inhibitor of Nrf2 (ML385), the promoter of Nrf2 (t-BHQ), and the inhibitor of ferroptosis (Liproxstatin-1), and the expression levels of autophagy-related proteins p62 and LC3, as well as Nrf2 and ferroptosis-related proteins xCT and GPX4 by WB, foam cell survival by CCK8, and intracellular reactive oxygen levels by Flow cytometry analysis and fluorescence microscopy. The effect of autophagy through Nrf2 on ferroptosis in foam cells was determined. The results revealed that insufficient autophagy in CQ-induced foam cells could lead to foam cell death in atherosclerotic plaques, and the cause of cell death was that insufficient autophagy in foam cells turned off the positive effect of Nfr2 antioxidant, initiated the negative effect of Nrf2 to promote intracellular reactive oxygen species production, and this negative effect promoted ferroptosis in foam cells.

Abstract Background: By associated with inflammation intraplaque, Trimethylamine N-oxide (TMAO) increase the risk of atherosclerotic plaque rupture and has been identified as the independent predictor of cardiovascular events. However, the underlying mechanism is yet unclarity. Accumulating studies have established the critical role of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome in mediating intraplaque inflammation and plaque progression. Here, we discussed the role of NLRP3 inflammasome in aggravating ox-LDL-induced macrophage inflammation response to TMAO and its potential mechanism. Results: Our results showed that TMAO enhanced ox-LDL-induced inflammation in THP-1cells. Adding to TMAO remarkably upregulated the expression or mRNA level of NLRP3, Cysteinyl aspartate specific proteinase 1(caspase-1) p20 and Apoptosis associated speck-like protein containing CARD(ASC) and enhanced the activity of caspase-1. An NLRP3 inhibitor (MCC950) reversed the promoting effect of NLRP3 inflammasome activation by TMAO and subsequently macrophage inflammation. Significantly, TMAO also boosted the activation of endoplasmic reticulum stress (ERS) and nuclear factor-kappa B (NF-kB) pathway in ox-LDL-induced cells, manifested as the increasing expression of p-NF-κB, Bip and phosphorylated protein kinase R-like ER kinase (p-PERK). Activation of the NLRP3 inflammasome by TMAO was reversed by the ERS inhibitor 4-PBA or the NF-κB phosphorylation inhibitor JSH-23. Meanwhile, 4-PBA further inhibited the NF-κB phosphorylation and alleviated the NLRP3 inflammasome activation. Conclusions: We concluded that TMAO exacerbates ox-LDL-induced NLRP3 inflammasomes activation and subsequently interleukin (IL)-18 and IL-1b release in THP-1 macrophages, which partly regulated by the activating of the PERK/NF-κB signaling pathway.

动脉粥样硬化是一种伴随脂质沉积、炎细胞浸润、泡沫细胞形成的慢性炎症性病理过程,是引发心脑血管疾病的重要病理基础.研究显示,粥样斑块的形成是动脉粥样硬化性疾病的基本病理特征,而颈动脉易损斑块的形成则是引发缺血性脑血管病的主要原因.本文主要综述了目前临床检测颈动脉易损斑块的常用方法,以期为临床诊疗提供参考.

Background. Systemic lupus erythematosus (SLE) is an autoimmune disease that can cause damage to multiple systems of the body. A number of studies have shown that long-chain noncoding RNA (lncRNA) can participate in the occurrence and development of a variety of autoimmune diseases. This study is aimed at detecting the expression levels of 5 lncRNAs in SLE patients and healthy controls and at exploring the relationship between expression levels and clinical symptoms and laboratory indicators. Methods. The design type of this study is a case-control study. A total of 76 SLE patients and 71 healthy controls were included in the first phase of the study. Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression level of 5 kinds of lncRNAs including lnc7514, lnc0640, lncagf, nc3643, and lnc5150 in PBMCs of two groups of patients; the expression of lncRNAs in the case group and the control group was analyzed. We analyzed the differences in the expression levels of lncRNAs between case and control groups, and explored the association of expression levels with clinical manifestations and laboratory characteristics. SPSS23.0 was used to analyze the expression level and gene polymorphism results; the statistical analysis test level α = 0.05 . Results. The expression level of lnc0640 in PBMCs of SLE patient group was higher than that of healthy control group ( Z = − 3.56 , P = 0.03 ). However, lnc5150 was lower than in healthy controls ( Z = − 7.16 , P < 0.001 ). lnc3643 expression levels were lower in SLE patients of SLE patients with pleurisy was lower than that of patients without pleurisy ( Z = − 2.44 , P = 0.02 ). Low lnc3643 expression levels were observed in PBMCs with SLE patients with rash symptoms ( Z = − 2.75 , P = 0.013 ). SLE expressed lower lnc3643 levels in PBMCs with SLE compared with those without pleurisy ( Z = − 2.42 , P = 0.02 ). The above differences were statistically significant. Association analysis of lncRNA expression levels and clinical manifestations in SLE patients found that SLE was lower than those without rash or pleurisy (both P < 0.05 ); association analysis of lncRNA expression level and laboratory results found a negative correlation between lnc3643, lnc7514, and SLE disease activity score (SLEDAI-2K), blood sink (ESR), and C-reactive protein (CRP) (all P < 0.05 ). Conclusions. lnc0640 was overexpressed in PBMCs in SLE patients compared with healthy controls. lnc3643 was negatively correlated with SLEDAI, and expression levels were associated with SLE patients with arthritis, rash, and pleuritis.