Abstract:Proprotein convertase subtilisin/kexin 9(PCSK9) is a serine protease which can induce the degradation of low density lipoprotein receptor(LDLR), increase the level of low density lipoprotein cholesterol(LDLC) in plasma. PCSK9 participate in various mechanisms promoting atherosclerosis. Recently, a number of clinical trials have confirmed that PCSK9 inhibitors have potential anti-inflammatory, anti-platelet, atherosclerotic plaque stability and other effects. This article reviews the research progress of PCSK9 and PCSK9 inhibitors from the aspects of the structure and regulatory factors of PCSK9, and the treatment of atherosclerosis by PCSK9 inhibitors.

Abstract:Acute coronary syndrome (ACS) is a group of clinical syndromes with high morbidity and mortality worldwide. Increasing evidence suggest that plaque erosion with an intact fibrous cap is one of the major causes responsible for ACS. Basic experiments have shed light on the unique molecular characteristics of plaque erosion. It has been indicated that plaque erosion starts with the changes of blood flow disturbance-induced endothelial cell damage, resulting in loss of basement membrane integrity and endothelial cell desquamation, with consequent formation of neutrophil extracellular traps and thrombosis. This review will discuss the molecular characteristics of atherosclerotic plaque erosion and translational research needed for future precision medicine in patients with plaque erosion.

Abstract:Atherosclerosis is a chronic inflammatory disease of artery. Abnormal activation of monocytes/macrophages is closely related to the development of atherosclerosis. Recent studies have found that monocytes/macrophages can undergo metabolic reprogramming by trained immunity, promote inflammatory responses, which accelerate the progression of atherosclerosis and increase cardiovascular incidents. In this review, we discuss the function and mechanisms of metabolic reprogramming of monocytes/macrophages in the trained immunity, which may provide new therapeutic targets and methods for the prevention and treatment of atherosclerosis.

Abstract:Aim To explore the effect of free fatty acid on the M1 polarization and glycolysis in mouse macrophages. The mechanism of non-alcoholic steatohepatitis (NASH) pathogenesis is clarified from the perspective of metabolism reprogramming and polarization in macrophages, which will provide a new way for prevention and treatment of NASH. Methods Oleic acid/palmitate acid (O/P) was used to treat RAW264.7 cells to establish cell model. AMPK-specific siRNA was transfected into RAW264.7 cells to knockdown AMPK protein expression. Acadesine (AICAR) was used to stimulate the activation of AMPK. The mRNA levels of M1 type molecular markers (TNF-α, IL-6 and MCP-1), M2 type molecular markers (CD206 and IL-10) and glycolysis related genes (PGM1, PGK1, GPI1, LDHA, ALDOA, GLUT1 and HK2) were detected by qPCR, and the protein levels of p-mTOR/mTOR, p-Raptor/Raptor, p-Akt/Akt, p-AMPK and AMPK were detected by Western blot. Results After treatment of RAW264.7 cells with O/P, the protein levels of p-mTOR/mTOR and p-Akt/Akt increased (P＜0.05), the protein levels of p-Raptor/Raptor, p-AMPK and AMPK decreased (P＜0.05), the mRNA levels of glucose glycolysis related genes PGM1, PGK1, GPI1, LDHA, ALDOA, GLUT1 and HK2 increased (P＜0.05), the mRNA levels of pro-inflammatory factors TNF-α, IL-6 and MCP-1 increased (P＜0.05), while the mRNA levels of anti-inflammatory factors CD206 and IL-10 decreased (P＜0.05). After inhibiting AMPK expression in RAW264.7 cells, the protein levels of p-mTOR/mTOR and p-Akt/Akt increased (P＜0.05), the protein levels of p-Raptor/Raptor, p-AMPK and AMPK decreased (P＜0.05), the mRNA levels of glucose glycolysis related genes PGM1, PGK1, GPI1, LDHA, ALDOA, GLUT1 and HK2 increased (P＜0.05), the mRNA levels of pro-inflammatory factors TNF-α, IL-6 and MCP-1 increased (P＜0.05), while the mRNA levels of anti-inflammatory factors CD206 and IL-10 decreased (P＜0.05). Activation of AMPK activity could reverse the effect of O/P on macrophage polarization and glycolysis. Conclusion Free fatty acid promotes M1 polarization and glycolysis via inhibiting AMPK pathway in mouse macrophages.

Abstract:Aim To explore whether inhibiting P53 and increasing the expression of glucose transporter 4 (GLUT4) in cardiomyocytes can improve glucose metabolism disorder and reduce apoptosis induced by high glucose (HG) combined with ischemia-hypoxia(IH). Methods The cardiomyocyte HG＋IH model was induced in vitro. The experimental groups were the control group, HG group, IH group, HG＋IH group, HG＋IH＋P53 inhibitor group (HG＋IH＋Pifithrin-α group), HG＋IH＋P53 inhibitor＋GLUT4 inhibitor group (HG＋IH＋Pifithrin-α＋Fasentin group). Cell viability was detected by the CCK-8 method, lactate dehydrogenase (LDH), glycolytic key enzyme activity, and ATP content were detected by the kit, protein expression of P53, GLUT4, Bax/Bcl-2 and Caspase-3 were detected by Western blot, and cardiomyocyte apoptosis was detected by flow cytometry. Results ①In the HG+IH cardiomyocyte model in vitro, compared with the control group, the expression of P53 increased by 75%, the expression of GLUT4 decreased by 16%, the content of ATP decreased by 51%, the cell viability decreased by 45%, the LDH activity increased by 3.6 times, the expression of Caspase-3 and Bax/Bcl-2 increased by 54% and 77% respectively, and the apoptosis rate increased (all P＜0.05). ②After inhibiting the expression of P53 in cardiomyocytes, compared with the HG＋IH group, the expression of GLUT4 in cardiomyocytes increased by 34%, the content of ATP increased by 60%, the cell viability increased by 50%, the LDH activity decreased by 13%, the expression of Caspase-3 and Bax/Bcl-2 decreased by 31% and 53% respectively, and the apoptosis rate decreased in HG＋IH＋Pifithrin-α group (all P＜0.05). ③After inhibiting GLUT4, compared with the HG＋IH＋Pifithrin-α group, the expression of GLUT4 in cardiomyocytes decreased by 22%, the content of ATP decreased by 39%, the cell viability decreased by 25%, the LDH activity increased by 21%, the expression of Caspase-3 and Bax/Bcl-2 increased by 43% and 89% respectively, and the apoptosis rate increased (all P＜0.05). Conclusions In the cardiomyocyte model of high glucose combined with ischemia-hypoxia, inhibiting P53 can increase the expression of GLUT4, improve the glucose metabolism disorder of cardiomyocytes induced by high glucose combined with ischemia-hypoxia, and reduce apoptosis.

Abstract:Atherosclerosis is a chronic inflammatory disease characterized by lipid deposition in the arterial vessel wall, in which both innate and adaptive immune cells are involved in the disease process and play different roles. Metabolic reprogramming of these immune cells modulates disease progression by modulating immune cell phenotype and function. This article reviews the metabolic pathway changes of major immune cells associated with the course of atherosclerosis, and summarizes the types and mechanisms of metabolic reprogramming among these immune cells.

Abstract:Aim To investigate the effect of dendrobium officinale polysaccharide (DOP) on Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in brain tissue of ischemic stroke rats. Methods The rat model of middle cerebral artery occlusion was established by thread embolism. SD rats were randomly divided into model group, DOP low dose (25 μg/g) group, DOP medium dose (50 μg/g) group, DOP high dose (100 μg/g) group and edaravone group, with 12 rats in each group; Another 12 SD rats were set as sham operation group. After drug intervention, the neurological deficit of rats in each group was scored by Longa grading method. The cerebral infarction in rats was detected by triphenyltetrazolium chloride staining. Hematoxylin-eosin staining was used to observe the pathological changes of hippocampal nerve tissue in rats. Enzyme-linked immunosorbent assay was used to detect the levels of inflammatory cytokines interferon-γ (IFN-γ), cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) in rat brain. Western blot was used to detect the expression level of JAK/STAT3 signaling pathway related proteins in rat brain. Results Compared with the sham operation group, the hippocampal neurons in the model group were shriveled, smaller, degenerated and necrotic, damaged in structure and disordered in arrangement, and the number of them decreased obviously. The whole hippocampal nerve tissue showed obvious pathological damage, and the neurological deficit score, cerebral infarction area, levels of IFN-γ, COX-2 and IL-6 in brain tissue, p-JAK/JAK, p-STAT3/STAT3 increased obviously (P＜0.05). Compared with the model group, the pathological damage of hippocampal nerve tissue in the drug treatment group was reduced, the neurological deficit score, cerebral infarction area, the levels of IFN-γ, COX-2 and IL-6 in brain tissue, p-JAK/JAK, p-STAT3/STAT3 were all decreased, and there was a dose-dependent relationship among DOP groups (P＜0.05). Compared with edaravone group, there was no significant change in all indexes of rats in DOP high dose group (P＞0.05). Conclusion DOP can inhibit the activation of JAK/STAT3 signaling pathway in ischemic stroke rats, reduce the synthesis and secretion of inflammatory cytokines, relieve brain inflammation and repair nerve function.

Abstract:Aim To investigate the influences of ginkgetin on the ferroptosis of vascular endothelial cells induced by oxidized low density lipoprotein (ox-LDL) by regulating the nuclear factor erythroid-2 related factor 2 (Nrf2)/solute carrier protein 7 family member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway. Methods Human umbilical vein cell fusion cells EA.hy926 were grouped into normal control group (normal culture), ox-LDL group (50 mg/L ox-LDL), and ginkgetin low dose group (50 mg/L ox-LDL+10 μmol/L ginkgetin), middle dose group (50 mg/L ox-LDL+20 μmol/L ginkgetin), high dose group (50 mg/L ox-LDL+40 μmol/L ginkgetin), ML385 group (50 mg/L ox-LDL+40 μmol/L ginkgetin+1 μmol/L Nrf2 inhibitor ML385), Erastin group (50 mg/L ox-LDL+40 μmol/L ginkgetin+5 μmol/L SLC7A11 inhibitor Erastin), RSL3 group (50 mg/L ox-LDL+40 μmol/L ginkgetin+0.5 μmol/L GPX4 inhibitor RSL3). Cell viability was detected by tetramethylazolium salt (MTT) method. The kits were applied to detect the levels of cellular superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH). The intracellular iron content was detected by specific fluorescent probe method. The levels of intracellular reactive oxygen species (ROS) and lipid ROS were detected by 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe method and boron dipyrrole (BODIPYTM) method; Western blot was applied to detect the protein expressions of cellular Nrf2, SLC7A11, GPX4,4-hydroxynonenoic acid (4-HNE), cyclooxygenase 2 (COX2), and p53. Results Compared with the normal control group, the cell viability, SOD content, GSH content, expressions of Nrf2, SLC7A11 and GPX4 were obviously decreased in the ox-LDL group (P＜0.05); the MDA content, Fe²＋ content, ROS, lipid ROS, expressions of 4-HNE, COX2, p53 were obviously increased (P＜0.05). Compared with the ox-LDL group, the cell viability, SOD content, GSH content, expressions of Nrf2, SLC7A11 and GPX4 were obviously increased in low, middle and high dose groups of ginkgetin (P＜0.05); the MDA content, Fe²＋ content, ROS, lipid ROS, expressions of 4-HNE, COX2, p53 were obviously decreased (P＜0.05). Compared with the high dose ginkgetin group, the ML385 group, Erastin group and RSL3 group attenuated the inhibitory effect of ginkgetin on ox-LDL-induced vascular endothelial cell ferroptosis. Conclusion Ginkgetin inhibits ox-LDL-induced vascular endothelial cell ferroptosis by activating the Nrf2/SLC7A11/GPX4 pathway.

Abstract:Aim To explore the influential factors of slow flow following rotational atherectomy (RA) and the predictive value of these factors. Methods According to the presence or absence of slow flow (defined as transient thrombolysis in myocardial infarction (TIMI) flow grade ≤2 just after RA), a total of 219 cases undergoing RA were divided into slow flow group (n=50) and non-slow flow group (n=169). The clinical history, laboratory examinations, and coronary intervention data of the two groups were compared. The two-class Logistic regression model was used to analyze independent influential factors of slow flow, the operating characteristic (ROC) curve was used to evaluate the predictive value. Results There were 50 patients (22.8%) in the slow flow group. Compared with the non-slow flow group, the final burr size of 1.25 mm was more used in the slow flow group (P＜0.05), while the final burr size of 1.5 mm was fewer (P＜0.05); Lesion length and total run time were longer, rotation times were higher (P＜0.05); while systolic blood pressure was lower in the slow flow group, and reference diameter was significantly smaller (P＜0.05). The two-class Logistic regression analysis showed that lesion length was an independent risk factor for slow flow, while reference diameter and systolic blood pressure were preventive factors (P＜0.05). Combined use of these variables provided incremental predictive value for slow flow after RA procedure, and the area under the curve was 0.736 with 78.0% sensitivity and 65.1% specificity. Conclusion Lesion length was an independent risk factor of slow flow, while reference diameter and systolic blood pressure were preventive factors, and the combination of these variables provided additional predictive value for slow flow in patients undergoing RA.

Abstract:Aim Construct a nomogram for predicting hospital mortality in critically ill patients with heart failure (HF) from Medical Information Mart for Intensive CareⅢ (MIMIC-Ⅲ) database. Methods Data were extracted involving critically ill patients with HF from MIMIC-Ⅲ database. All eligible patients (n=8 604) were randomly classified into the training set (n=6 022) and validation set (n=2 582) with a ratio of 7∶3, and the outcome was hospital mortality. LASSO-Logistic analysis in the training set was used to determine the prognostic factors and the nomogram for predicting hospital mortality was thereby constructed. Receiver operating characteristic (ROC) curve, calibration curve, decision analysis curve (DCA) and clinical impact curve (CIC) were generated to assess the discrimination, calibration and clinical utility of the nomogram, respectively. Results LASSO-Logistic analysis showed that red blood cell distribution width (RDW), respiratory rate, oxygen saturation, acute physiology score Ⅲ (APSⅢ) and simplified acute physiology score Ⅱ (SAPSⅡ) were independent predictors. In both training and validation set, the area under the curve (AUC) was 0.775 (95%CI:0.757～0.792) and 0.767 (95%CI:0.742～0.793), respectively. Calibration curve was highly consistent with the diagonal line, and the mean absolute error was 0.009 and 0.016. AUC and calibration curve showed great performance of the predictive model in discrimination and calibration. Meanwhile, DCA and CIC revealed that the predictive model provided significant net benefits for most threshold probabilities. Conclusion Nomogram is a simple and accurate tool for predicting hospital mortality in critically ill patients with HF.

Abstract:Aim To explore the relationship between fractional flow reserve (FFR) technical parameters and myocardial zymogram in patients with non-ST-segment elevation acute myocardial infarction (NSTEMI) and the predictive value of major adverse cardiovascular events (MACE). Methods A total of 136 NSTEMI patients admitted to Liaocheng Third People's Hospital from February 2017 to February 2020 were selected and all underwent percutaneous coronary intervention (PCI) under the guidance of FFR. According to whether MACE occurred 1 month after operation, all patients were divided into MACE group (n＝14) and non-MACE group (n＝122). The clinical data, myocardial enzyme spectrum indexes, quantitative coronary angiography (QCA) parameters, and FFR technical parameters were compared between the two groups. The correlation between FFR technical parameters and myocardial enzyme spectrum indexes and QCA parameters was analyzed. The Logistic regression equation was used to analyze MACE influencing factors. The receiver operating characteristic (ROC) curve and the area under curve (AUC) were used to analyze the myocardial zymogram indexes, QCA parameters, and FFR to predict the value of MACE. Results Compared with the non-MACE group, the proportion of patients aged ≥60 years old in the MACE group increased 0.55 times, the GRACE score increased 0.11 times, the cardiac troponin I (cTnI) level increased 0.38 times, the creatine kinase (CK) level increased 0.22 times, the creatine kinase isoenzyme-MB (CK-MB) level increased 0.24 times, and the stenosis degree increased 0.08 times, the lesion length increased 0.11 times, the FFR decreased 11.9%, and the minimum lumen area of criminal vessels decreased 12.8%, the difference was statistically significant (P＜0.05); FFR was negatively correlated with cTnI, CK, and CK-MB levels (P＜0.05); FFR was negatively correlated with stenosis and lesion length (P＜0.05); Logistic regression analysis showed that after adjusting for age, GRACE score, and culprit vessel minimum lumen area, myocardial zymogram indexes (cTnI, CK, CK-MB), QCA parameters (degree of stenosis, lesion length), and FFR were all related to the occurrence of MACE (P＜0.05); The AUC of MACE predicted by FFR was 0.885, the sensitivity was 92.86%, and the specificity was 65.88%. Conclusion There is a negative correlation between FFR and myocardial enzymes (cTnI, CK and CK-MB) levels in NSTEMI patients, and are related to the occurrence of MACE, which can predict the risk of MACE.

Abstract:Macrophages are heterogeneous and plastic, and the microenvironment is a key factor affecting the phenotypic changes of macrophages. In cardiovascular disease, macrophages can polarize into different phenotypes (M1, M2,et al) depending on their microenvironment. They express different surface receptors, release different cytokines, are regulated by a variety of signaling pathways and transcriptions, and perform different functions in the development of cardiovascular diseases. M1 is responsible for initiating and maintaining inflammatory responses by secreting pro-inflammatory factors; M2 is involved in inflammation resolution and tissue repair by secreting anti-inflammatory factors. Traditional Chinese medicines have the functions of strengthening the body, promoting blood circulation and removing blood stasis, clearing heat and detoxifying. By regulating JAK/STAT, PI3K/Akt, Toll receptors and other signaling pathways, it reduces the aggregation of macrophages, promotes the polarization of macrophages to M2 or restores M1/M2 homeostasis to improve the inflammatory microenvironment and delay disease progression. This paper mainly systematically sorts out the origin of macrophages, the polarization and regulation mechanism of macrophages, the role of macrophage polarization in cardiovascular diseases and the intervention of traditional Chinese medicine, and provides theoretical guidance for clinical prevention and treatment of cardiovascular diseases.

Abstract:N6-methylpurine (m⁶A) is the most common type of post-transcriptional RNA modification in eukaryotes, involving many types of RNA. m⁶A methylation modification is dynamic and reversible and is mainly regulated by a variety of enzymes and proteins, including methyltransferase, demethylase and m⁶A-related binding proteins. Atherosclerosis is the main cause of cardiovascular and cerebrovascular diseases. Recent studies have found that m⁶A methylation is closely related to atherosclerosis. This paper summarizes the current understanding of the mechanism of m⁶A methylation modification, and reviews the mechanism and latest progress of m⁶A methylation modification in atherosclerosis-related cells, so as to provide a new target for the diagnosis, prevention and treatment of atherosclerosis.