Abstract:

Liquid-liquid phase separation is an important process in which intracellular biomolecules condense to form membraneless organelles, playing a significant role in cell signaling and gene expression regulation. Studies have shown that liquid-liquid phase separation may be involved in the occurrence and development of atherosclerosis and has potential applications in the diagnosis and treatment of the disease. This article introduces the concept and mechanism of liquid-liquid phase separation, delves into the relationship between liquid-liquid phase separation and atherosclerosis, summarizes the progress of liquid-liquid phase separation in atherosclerosis research, and looks forward to its application prospects.

Abstract:

The gut microbiome actively regulates host immunity, digestive processes, and the function of the intestinal endocrine system. Additionally, it modulates host neural signal transmission and nutrient metabolism through metabolite generation. Vascular calcification involves the deposition of calcium phosphate in blood vessel walls, secondary to metabolic disorders such as chronic kidney disease, atherosclerosis, diabetes, and osteoporosis. Recent research spanning the past two decades has indicated a close correlation between shifts in the composition and functionality of the gut microbiota, along with its metabolites, and the onset of metabolic disease-related vascular calcification. This paper presents a comprehensive review of the roles and mechanisms of the gut microbiota in this context.

Abstract:

Aim To study the diastolic effect and mechanism of farrerol on isolated pulmonary arteries of C57BL/6J mice. Methods After anesthesia, mouse lung tissue was quickly removed and placed into the 4 ℃ K-H buffer, pulmonary arteries were isolated under the microscope and cut into 2 mm long vascular rings for spare use. (1)The effect of farrerol on the resting tension of isolated mouse pulmonary arteries:in the resting state, the active mouse pulmonary artery rings were treated with different concentrations of farrerol (10－6,3×10－6,0－5,3×10－5 and 10－4 mol/L). (2)Farrerol relaxed mouse pulmonary artery experiment:pulmonary arteries were contracted using phenylephrine (PE, 1 μmol/L) or KCl (60 mmol/L), and when the contraction reached the platform, different concentrations of farrerol (10－6,3×10－6,0－5,3×10－5 and 10－4 mol/L) was added. (3) Farrerol inhibited pulmonary artery contraction experiment:under conditions with or without the addition of farrerol, pulmonary arteries were contracted using different concentrations of PE (10－9,3×10－9,0－8,3×10－8,0－7,3×10－7 and 10－6 mol/L) or KCl (0,0, 0,0, 80 and 120 mmol/L), and the pulmonary artery muscle tension was recorded. (4)Calcium free and recalcification experiments:under conditions with or without the addition of farrerol, the changes of isolated mouse pulmonary artery tension were measured in the state of calcium free or recalcification {2.5 mmol/L [Ca²＋]ex}. (5)The relationship between farrerol induced relaxation of isolated mouse pulmonary arteries and potassium ion channels:firstly, 60 mmol/L KCl solution was used to contract the mouse pulmonary arteries until the platform. Then, 3 mmol/L aminopyridine (4-AP), 2 mmol/L tetraethylammonium (TEA), 30 μmol/L BaCl₂, and 10 μmol/L glibenclamide (Gli) were added and treated for 15 min. Subsequently, the pulmonary arteries were relaxed using a concentration gradient of farrerol. Results Farrerol had no significant effect on the mouse pulmonary arteries in the resting state, but had a concentration-dependent relaxing effect on the mouse pulmonary arteries pre-contracted with PE and KCl. While the pretreatment of 3×10－5 mol/L farrerol could significantly reduce the maximum contraction of mouse pulmonary arteries induced by PE and KCl (P＜0.01), as well as significantly reduce the contraction of mouse pulmonary arteries induced by KCl under calcium free or recalcification conditions (P＜0.01). Addition of the voltage-dependent potassium ion channel blocker 4-AP significantly reduced the maximum diastolic rate of mouse pulmonary arteries induced by farrerol (P＜0.01), while addition of the high conductivity calcium activated potassium ion channel blocker TEA, inward rectifying potassium ion channel blocker BaCl₂, or ATP sensitive potassium ion channel blocker Gli had no significant effect on the vasodilation effect of farrerol (P＞0.05). ConclusionFarrerol has a relaxing effect on isolated mouse pulmonary arteries, and its mechanism may be related to open voltage-dependent potassium ion channels.

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 investigate the effect of hypobaric hypoxia on macrophage necroptosis and atherosclerotic plaque instability and explore the underlying mechanisms. Methods Mouse bone marrow-derived macrophages were isolated and cultured, and divided into control group (21% oxygen concentration) and hypoxia group (3% oxygen concentration). After 48 hours, cell necroptosis was detected, and the expression of cell necroptosis related proteins was determined by Western blot. Healthy male ApoE－/－ mice were randomly divided into control group and hypobaric hypoxia group. After the intervention for 16 weeks, the plasma lipids and inflammatory cytokines were measured, the areas of atherosclerotic plaque and necrotic core were evaluated by HE staining. The content of plaque collagen was detected by Masson staining. The number of macrophages in the plaque and the expression of necrotic apoptosis related proteins were detected by immunohistochemical staining and Western blot. Results Hypoxia induced increased necrotic apoptosis of macrophages (P＜0.01), while necroptotic inhibitor necrostatin-1 (Nec-1) reduced hypoxia induced cell death (P＜0.05); hypoxia leads to a decrease in the expression of adenosine deaminase acting on RNA 1 (ADAR1) in macrophages (P＜0.01), and an increase in the expression of Z-DNA binding protein 1 (ZBP1), phosphorylated receptor-interacting serine/threonine-protein kinase (p-RIPK3), and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) (all P＜0.01). Compared with the control group, the plasma lipid levels of ApoE－/－ mice in the hypobaric hypoxia group did not change significantly (P＞0.05), the plasma inflammatory cytokines (TNF-α, IL-1β, IL-6 and MCP-1) increased (all P＜0.05), the area of atherosclerotic plaque increased (P＜0.05), the area of plaque necrotic core increased, the content of plaque collagen decreased, the number of macrophages increased, the expression of ADAR1 decreased, and the expression of ZBP1 and p-MLKL increased (all P＜0.01). Conclusion Hypobaric hypoxia causes the imbalance of ADAR1/ZBP1 expression in macrophages, activates RIPK3/MLKL signaling pathway, promotes macrophage necroptosis, increases the area of plaque necrosis core, and leads to increase instability of atherosclerotic plaque.

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:

Aim To apply coronary angiography derived index of microcirculatory resistance (caIMR) to evaluate the effect of coronary microcirculation perfusion on myocardial remodeling after interventional therapy in patients with acute anterior ST segment elevation myocardial infarction (STEMI). Methods This was a cross-sectional study.The analysis was performed among the patients who were hospitalized for acute anterior STEMI in the First Department of the Second Affiliated Hospital of Dalian Medical University from January 2021 to July 2022 and received percutaneous coronary intervention (PCI) with regtelar follow-up visits. The patients were divided into low caIMR (L-caIMR) group, medium caIMR (M-caIMR) group and high caIMR (H-caIMR) group according to the results of caIMR. The results of echocardiography at perioperative period, 1 month, 3 months, 6 months and 1 year were analyzed and compared, including left atrial diameter (LAD), left ventricular end-diastolic diameter (LVEDD), interventricular septum thickness (IVST), mitral orifice flow velocity E/A, mitral annular septum e′ and mitral annular wall e′, etc. The difference of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and other inflammatory factors in peripheral blood of the three groups were also compared. Results A total of 75 patients diagnosed with acute anterior STEMI were recruited, including 55 males. The L-caIMR group, M-caIMR group, and H-caIMR group had 6,6 and 23 cases, respectively. Compared with the L-caIMR group, the LAD and IVST in the M-caIMR group and the H-caIMR group exhibited an increasing tendency one month after PCI, and the increase in the H-caIMR group was more significant than that in the M-caIMR group (P＜0.05). The ejection fraction in the H-caIMR group was notably lower than that in the L-caIMR group and the M-caIMR group at 1 and 3 months after PCI (P＜0.05). Compared with the L-caIMR group, the mitral flow velocity E/A at 6 months after PCI, and the e′ at the septal side and the lateral wall of the mitral annulus at 1,3, and 6 months after PCI were significantly reduced in the M-caIMR and H-caIMR groups(P＜0.05). Compared with the L-caIMR group, the levels of IL-1β, IL-6, and TNF-α showed an increasing trend in the M-caIMR group and the H-caIMR group, and the increase was greater in the H-caIMR group than that in the M-caIMR group (P＜0.05). Multivariate analysis revealed that caIMR was a factor influencing the levels of IL-1 β and IL-6 (P＜0.05). Conclusion CMD may be involved in the process of myocardial remodeling in patients with acute anterior STEMI after PCI, in which inflammation plays a role.

Abstract:

Aim To detect the changes of serum cardiotrophin-1 (CT-1) and angiopoietin-like protein 3 (ANGPTL3) levels in patients with coronary heart disease (CHD) complicated with heart failure (HF) after percutaneous coronary intervention (PCI), and analyze their relationship with prognosis. Methods 199 patients with CHD complicated with HF who underwent PCI in the Second Affiliated Hospital of Zhengzhou University from March 2022 to March 2023 were selected. The serum CT-1 and ANGPTL3 levels of patients with different New York Heart Association (NYHA) cardiac function grades were compared before surgery. The prognosis was followed up after PCI, and the patients who had major adverse cardiovascular event (MACE) were included in the poor prognosis group, and the rest were included in the good prognosis group. The general data and serum CT-1 and ANGPTL3 levels were compared between the poor prognosis group and the good prognosis group. Logistic regression model was used to analyze the influencing factors of poor prognosis after surgery in patients with CHD and HF. The predictive value of serum CT-1 and ANGPTL3 alone and in combination were analyzed. Results Compared with the patients with cardiac function grade Ⅰ, the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅱ, Ⅲ and Ⅳ were increased (P＜0.05). Compared with the patients with cardiac function grade Ⅱ, the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅲ and Ⅳ were increased (P＜0.05). Compared with the patients with cardiac function grade Ⅲ, the serum CT-1 and ANGPTL3 levels of the patients with acrdiac function grade Ⅳ were increased (P＜0.05). Spearman correlation analysis showed that the serum CT-1 and ANGPTL3 levels were positively correlated with NYHA cardiac function grade (r＝0.8,5%CI:0.408~0.613, P＜0.001, r＝0.7,5%CI:0.666~0.794, P＜0.001). The poor prognosis rate of patients was 17.93%. Compared with the good prognosis group, the serum CT-1 and ANGPTL3 levels of the poor prognosis group were increased (P＜0.05). Logistic regression model analysis showed that smoking, diabetes, lesion vessel number≥3, irregular medication outside the hospital, serum CT-1 and ANGPTL3 levels were the influencing factors of poor prognosis in patients with CHD complicated with HF (P＜0.05). ROC curve analysis showed that the sensitivity and area under the curve (AUC) of combined serum CT-1 and ANGPTL3 levels for predicting poor prognosis of patients with CHD complicated with HF were higher than those of either marker alone, while the specificity was basically similar to that of single-marker prediction. Conclusion Serum CT-1 and ANGPTL3 levels are abnormally elevated in patients with CHD complicated with HF after PCI, and are closely related to the cardiac function and prognosis.

Abstract:

Renal arterial stenosis (RAS) hypertension is one of the common types of secondary hypertension. The main clinical manifestations are increased levels of renin and aldosterone, abnormal renal function and refractory hypertension that is difficult to control by drugs. In this paper, a case with refractory hypertension due to atherosclerotic renal artery stenosis was reported and the relevant literature was reviewed. The patient was a middle-aged male who could not control his blood pressure within the ideal range despite oral administration of amlodipinebesylate, terazosinhydrochloride, irbesartan, carvedilol, metoprololsuccinate, diltiazium hydrochloride and other drugs, requiring continuous pumping of nitroglycerin to lower blood pressure. Furthermore, the examination of renal artery angiography revealed severe stenosis at the origin of the left renal artery. After stent implantation at the renal artery stenosis, blood pressure could be controlled to the standard by oral administration of terazosin hydrochloride, amlodipine besylate and diltiazem hydrochloride only. There are many reasons leading to secondary hypertension. This paper discusses the common causes of renal artery stenosis as a starting point, in order to deepen the understanding of secondary hypertension and reduce the occurrence of missed diagnosis or misdiagnosis.

Abstract:

N⁶-methyladenosine (m⁶A) is the most common mRNA modification in eukaryotes, and fat mass and obesity-related protein (FTO), are its demethylases, which efficiently remove the modification of m⁶A mRNA, and is strongly associated with obesity. Atherosclerosis is a chronic inflammatory lesion of the blood vessel wall driven by lipids.It was found that FTO-mediated m⁶A may influence the process of atherosclerosis through lipid metabolism, oxidative stress, mitochondrial dysfunction, and macrophage foaminess.

Abstract:

Diabetes mellitus type 2 (T2DM) is a common chronic metabolic disease, compared to non-diabetics, T2DM patients have a higher risk of heart disease, and their risk of vascular diseases is twice that of non-diabetic individuals. Multiple clinical trials have confirmed that glucagon-like peptide-1 receptor agonists (GLP-1RA), in addition to their function of lowering blood glucose, can also decrease the risk of cardiovascular events in T2DM patients. This article reviews the mechanisms of GLP-1RA-mediated cardioprotection. It provides a comprehensive review of the recent progress in GLP-1RA-mediated cardiac protective mechanisms, elaborating on the protective effects of GLP-1RA on specific heart cell and further discussing its impact on heart failure (HF), provide assistance for clinical treatment of diabetic cardiomyopathy.

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.