Abstract:Ferroptosis is a form of programmed cell death, which is characterized by the accumulation of ferrous irons and a large number of lipid peroxides in cells. Metabolic cardiovascular diseases are characterized by cardiovascular dysfunction involving a series of metabolic factors. More and more studies have shown that ferroptosis plays an important role in the occurrence and development of metabolic cardiovascular diseases. Therefore, exploring the role of ferroptosis in metabolic cardiovascular diseases has vital clinical significance for revealing the pathogenesis and prevention strategies of metabolic cardiovascular diseases.

Abstract:Aim To investigate the relationship between apolipoprotein E (ApoE) receptor 2 (ApoER2) degradation by proprotein convertase subtilisin kexin 9 (PCSK9) and the anti-inflammatory effect of ApoE/ApoER2. Methods Human umbilical vein endothelial cells (HUVEC) and HepG2 cells were cultured in vitro, Western blot and ELISA were used to detect the effects of lipopolysaccharide (LPS) on the expression and secretion of Toll-like receptor (TLR4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and PCSK9 in HUVEC; the effects of ApoE3 on the expression and secretion of TNF-α, IL-6, PCSK9 and ApoER2 in HUVEC induced by LPS; the effects of three isoforms of ApoE (ApoE2, ApoE3 and ApoE4) on the expression of PCSK9 and ApoER2 in HUVEC and HepG2 cells under non-inflammatory conditions; and the effects of PCSK9 on the expression and secretion of ApoER2, TNF-α and IL-6 in HUVEC. Results Western blot and ELISA showed that LPS up-regulated the expression and secretion of TLR4, TNF-α, IL-6 and PCSK9 in HUVEC; ApoE3 inhibited LPS-induced inflammatory responses, and up-regulated ApoER2 expression and secretion; the three isoforms of ApoE (ApoE2, ApoE3 and ApoE4) had no effect on the expression of PCSK9 and ApoER2 in HUVEC and HepG2 cells under non-inflammatory conditions. Different doses (0,0.5,1.0 and 2.5 mg/L) of recombinant human PCSK9 were used to treat HUVEC for 24 h, Western blot and ELISA showed that PCSK9 up-regulated the expression and secretion of TNF-α and IL-6, and down-regulated the expression of ApoER2. Conclusion PCSK9 antagonizes the anti-inflammatory effects of ApoE/ApoER2 by degrading ApoER2.

Abstract:Aim To investigate the protective effect and mechanism of apigenin (APG) on cognitive function in vascular dementia (VD) rats. Methods The VD rat model was replicated by ligating bilateral common carotid arteries, and the sham group, model group, APG low-dose group, APG high-dose group, nimodipine (NMP) group were setted, with 30 rats in each group. The rats in APG low-dose group and high-dose group were administered intraperitoneally once a day at doses of 0,0 mg/kg APG respectively, the rats in the NMP group were administered intraperitoneally once a day at doses of 2 mg/kg NMP, the rats in the sham group and model group were given normal saline, the course of treatment was 28 days. The cognitive function was detected by Morris water maze test, the pathological characteristics and apoptosis of neurons in the CA1 region of the hippocampus were observed through HE staining and TUNEL staining, the ultrastructural changes of neurons was observed by electron microscopy. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in hippocampal were detected by spectrophotometric method. The mRNA and protein expression of nuclear factor E2 related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by RT-PCR and Western blot. Results Compared with the model group, the cognitive function was significantly improved of rats in the APG high-dose group and NMP group; the number of neurons of hippocampus CA1 area, morphological and structural pathological changes, ultra-micromorphological changes and apoptosis were significantly improved, the pathological grade and apoptosis index (AI) were reduced; the content of MDA in the hippocampus was decreased, and the activity of SOD, GSH-Px were increased; the mRNA and protein expression of Nrf2, HO-1 were increased. The APG high-dose group had better effects on all indicators than those of NMP group. ConclusionAPG can significantly improve the cognitive function of VD rats, which may be related to the activating Nrf2/HO-1 pathway by APG, inhibiting oxidative stress and reducing brain hippocampus damage.

Abstract:Aim To observe the effect of edaravone dexborneol (EDDE) on neuronal apoptosis in rats with cerebral ischemia-reperfusion, and to explore its mechanism. Methods SD rats were randomly divided into sham group, middle cerebral artery occlusion (MCAO) group, low-dose EDDE group (EDDE-L group, 3 mg/kg), and high-dose EDDE group (EDDE-H group, 6 mg/kg), chelerythrine (CHE, PKC inhibitor) group (5 mg/kg CHE), EDDE＋CHE group (6 mg/kg EDDE＋5 mg/kg CHE), with 15 rats in each group. Except for the sham group, the rats in the other groups were given the suture method to construct the MCAO model. The rats in each group were scored for neurological deficits; after 24 hours of reperfusion, TTC staining was used to detect the cerebral infarct volume of rats; HE staining was used to observe the pathological damage of cortical nerve cells; TUNEL staining was used to detect cortical nerve cells apoptosis; immunohistochemical method was used to detect the expression of Bcl-2 and Bax in the cortex nerve cells; Western blot was used to detect the expression of Caspase-3, cleaved Caspase-3 and protein kinase C (PKC)/extracellular signal-regulated kinase (ERK) pathway related proteins (PKC, p-PKC, ERK1/2 and p-ERK1/2) in brain tissue. ResultsCompared with sham group, the neurological deficit score, the percentage of cerebral infarction volume and the apoptosis rate of nerve cells increased, the positive expression of Bax increased and the ratio of cleaved Caspase-3/Caspase-3 increased (all P＜0.05), the positive expression of Bcl-2 decreased, the ratio of Bcl-2/Bax, p-PKC/PKC and p-ERK1/2/ERK1/2 decreased in brain tissue (all P＜0.05), the cerebral cortex cells were sparsely arranged, swelling and vacuolar degeneration of nerve cells were obvious, the nucleus shrank and the tissue becomes necrotic in MCAO group. Compared with MCAO group, the neurological deficit score, cerebral infarction volume percentage and apoptosis rate of nerve cells decreased, the positive expression of Bax decreased, the ratio of cleaved Caspase-3/Caspase-3 decreased (all P＜0.05), the positive expression of Bcl-2 increased, the ratio of Bcl-2/Bax, p-PKC/PKC and the ratio of p-ERK1/2/ERK1/2 increased in brain tissue (P＜0.05) in the EDDE-L group and EDDE-H group, the pathological damage of the cerebral cortex was improved to varying degrees, the number of nerve cells increased, the vacuolar degeneration was reduced, the nucleus was clearer, and the EDDE-H group was better than the EDDE-L group. CHE could eliminate the neuroprotective effect of EDDE. Conclusion EDDE cloud inhibit neuronal apoptosis and reduce cerebral ischemia/reperfusion injury, and its mechanism may be related to the activation of PKC/ERK pathway.

Abstract:Aim To investigate the antagonistic effect and mechanism of amarogentin (MAG) on atherosclerosis induced by high fat diet (HFD) in mice. Methods Thirty ApoE－/－mice were randomly divided into control group (normal diet), HFD group and HFD+MAG group, with 10 mice in each group. HFD+MAG group was given 50 mg/kg MAG daily for 12 weeks. Blood lipid level was measured by biochemical detector. The deposition of plaque in aorta was observed by oil red O staining. The pathological morphology of aorta was observed by hematoxylin-eosin (HE) staining.Mac-3 immunohistochemistry was used to observe the aggregation of macrophages in aorta. The expression and distribution of NOD-like receptor pyrin domain containing 3 (NLRP3) were observed by immunofluorescence location staining. The expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and Caspase-1 subunit p20 in aorta were detected by Western blot. Results The levels of triglyceride, total cholesterol and low-density lipoprotein cholesterol in HFD+MAG group were respectively decreased to 88.39%, 74.85% and 64.97% of HFD group (P＜0.05), and the level of high-density lipoprotein cholesterol was increased to 142.18% of HFD group (P＜0.05), the plaque area was reduced to 45.16% of HFD group (P＜0.05), the pathological morphology of aorta was improved, macrophage deposition and the proportion of NLRP3 positive macrophages in plaque was reduced, and the expression levels of NLRP3, ASC and Caspase-1 subunit p20 in aorta were respectively decreased to 45.58%, 53.28% and 42.35% of HFD group (P＜0.05). Conclusion MAG has significant antagonistic effects on atherosclerosis mice induced by high fat diet, which may be related to the inhibition of NLRP3 inflammasome activation of macrophages in plaques.

Abstract:Aim To explore the mediating effect of Th17/Treg cell functional axis on arteriosclerosis induced by depressive stress in older adults. Methods From May 2018 to August 9,9 older individuals aged 60 and above were enrolled in community in Jinan area, Shandong. Geriatric depression scale (GDS) was used to evaluate depressive stress of the participants. According to the quintile of the GDS scores, participants were divided into low depressive stress group (n=349) and high depressive stress group (n=190). Common carotid intima-media thickness (CCA-IMT) and carotid-femoral pulse wave velocity (cfPWV) were used to assess arteriosclerosis and stiffness. The serum level of high sensitivity C-reactive protein (hs-CRP) and the frequencies of peripheral blood Th17 and Treg cells and relative cytokines were detected. Results Compared with the low depressive stress group, CCA-IMT, cfPWV, Th17 cell frequencies, the levels of hs-CRP, interleukin-17 (IL-17), tumor necrosis factor alpha (TNF-α), IL-6 and IL-23, the expression of retinoic acid receptor-related orphan receptoryγt (RORγt) mRNA, and the ratio of Th17/Treg cells in the high depressive stress group increased by 24.58%, 18.22%, 51.59%, 44.79%, 77.60%, 55.94%, 61.49%, 41.13%, 72.11% and 150.00% respectively; while the Treg cell frequencies, the levels of IL-10 and transforming growth factor β1 (TGF-β1), and the expression of forkhead box protein P3 (Foxp3) mRNA decreased by 36.53%, 32.96%, 27.65% and 35.35% respectively (all P＜0.001). After adjusting for confounding factors, GDS score was independently and positively correlated with CCA-IMT, cfPWV, Th17 cell frequencies, hs-CRP, IL-17, TNF-α, IL-6, IL-23, RORγt mRNA expression, and Th17/Treg cells ratio, whereas it was independently negatively correlated with Treg cell frequencies, IL-10, TGF-β1, and Foxp3 mRNA expression (all P＜0.001). The results of Hayes Process analysis showed that hs-CRP, Th17/Treg cells ratio, IL-17, and IL-10 played significant mediating roles in the correlations of GDS with CCA-IMT and cfPWV. Conclusions Depressive stress is an independent risk factor for arteriosclerosis in the elderly. Chronic inflammation may be an important mediating effector in the process of arteriosclerosis induced by depressive stress.

Abstract:Aim To explore the influencing factors of fat attenuation index of pericoronary adipose tissue (FAI-PCAT) when there is no local inflammation or mild inflammation of coronary artery, and to establish a regression model to predict the individual baseline values of FAI-PCAT. Methods Hospitalized patients who had no previous history of coronary heart disease and underwent coronary computed tomography angiography (CCTA) examination to exclude obstructive coronary artery disease in the First Affiliated Hospital of Harbin Medical University from January to December 2019 were included. The baseline data and clinical indexes such as sex, age, height, weight, past medical history, blood routine, blood glucose, and blood lipid were collected. The AW VolumeShare 4 CT working software was used to post-process the CCTA images by one doctor with more than 3 years of imaging experience, and FAI-PCAT and epicardial fat attenuation (EFat) were measured and recorded. Spearman correlation was used to analyze the correlation of factors. Multiple linear regression was used to analyze the influencing factors of FAI-PCAT and then established a regression model. Results A total of 103 patients with CCTA showing CAD-RADS 0～1 were included. The FAI-PCAT value is (－82.2±7.0) Hu, the EFat value is (－84.8±4.4) Hu. Correlation analysis showed that FAI-PCAT was significantly and positively correlated with EFat (P＜0.01), significantly and negatively correlated with lymphocyte to monocyte ratio (LMR), lymphocyte percentage (P＜0.01), and positively correlated with blood glucose level (P＜0.05). There was no correlation between FAI-PCAT and body mass index (BMI), blood lipid. The EFat was significantly and negatively correlated with BMI, triglyceride (TG), low density lipoprotein cholesterol/high density lipoprotein cholesterol (LDLC/HDLC), very low density lipoprotein cholesterol (VLDLC) (P＜0.01), and significantly and positively correlated with HDLC (P＜0.01), positively correlated with apolipoprotein A/apolipoprotein B (ApoA/ApoB) (P＜0.05). Multiple linear regression analysis showed that EFat value (X₁), LMR (X₂), and blood glucose level (X₃) were the main influencing factors and prediction parameters of FAI-PCAT value when there was no local inflammation or mild inflammation of coronary artery, and the regression equation was =－25.466+0.686X₁－0.9X₂₊₁.207X₃. Conclusion EFat value, LMR, and blood glucose level were the main influencing factors of FAI-PCAT value when there was no local inflammation or mild inflammation of coronary artery, and the regression model can predict the individual baseline values of FAI-PCAT.

Abstract:Aim To explore the potential causal association between galectin-1 (Gal-1) levels and atherosclerosis (As). Methods Single nucleotide polymorphisms (SNP) associated with Gal-1 served as instrument variables (IV), and the causal association between genetically predicted Gal-1 levels and As was analyzed by the two-sample Mendelian randomization (MR) method. Results Inverse variance weighted (IVW) results showed that genetically predicted Gal-1 levels were positively associated with risk of peripheral As and other As type (excluding cerebral artery, coronary artery, and peripheral artery) after Bonferroni adjustment (OR=1.6,5%CI:1.05～1.27, P＝0.002; OR=1.6,5%CI:1.12～1.20, P＝ 4.11E－17). There was no evidence supporting the causal association between Gal-1 and either coronary As or cerebral As (OR=1.2,5%CI:0.91～1.14, P＝0.765; OR=1.0,5%CI:0.94～1.29, P＝0.220); After Meta-analyzed the MR estimates of As outcomes at different sites, the results showed that genetically predicted Gal-1 levels were positively associated with As risk (OR=1.2,5%CI:1.06～1.19). ConclusionThe study suggests that genetically predicted Gal-1 level is causally associated with As risk, and Gal-1 is a potential target to prevent the occurrence of As.

Abstract:Aim To explore the clinical value of transcutaneous oxygen pressure (TcPO₂) combined with serum oxidized low density lipoprotein (ox-LDL) in predicting restenosis after interventional therapy for lower arteriosclerosis obliterans (LASO). Methods 113 patients with LASO underwent interventional therapy in Yueyang Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2020 to June 2021 were enrolled, and classified into non-stenosis group (n＝79) and restenosis group (n＝34) according to the occurrence of restenosis 1 year after interventional treatment. Serum ox-LDL levels were detected by ELISA and corresponding assay kits, and TcPO₂ was measured by laser Doppler flowmeter. Then comparison was conducted on general data, TcPO₂ and ox-LDL. Multivariate Logistic regression model was used to screen the influencing factors of restenosis after intervention therapy for LASO. Spearman test was used for correlation analysis, ROC curve was used to evaluate the efficacy of TcPO₂ and ox-LDL in predicting restenosis in LASO patients after intervention therapy. Results There was no significant difference between the two groups in age, body mass index (BMI), sex, alcohol consumption, history of cerebrovascular disease and coronary heart disease, number of diseased vessels, fasting blood glucose, and homocysteine (P＞0.05). Compared with the non-stenosis group, smoking, irregular drug use, the number of implanted stents, ox-LDL, and the level of blood uric acid in the restenosis group were significantly higher (P＜0.05), and TcPO₂ was significantly lower (P＜0.05). Logistic regression model analysis showed that smoking, irregular drug use after LASO intervention, large number of implanted stents, decreased TcPO₂, ox-LDL, and increased blood uric acid were the risk factors for restenosis after LASO intervention (P＜0.05). Spearman test showed that ox-LDL was positively correlated with restenosis after LASO intervention (r＝0.513, P＜0.001), and TcPO₂ was negatively correlated with it (r＝－0.524, P＜0.001). The ROC curve showed that the predictive efficacy of TcPO₂＋ox-LDL in predicting restenosis after LASO interventional therapy (AUC＝0.802) was higher than that of each indicator alone. The predictive sensitivity and specificity were 67.60% and 94.90% respectively, and the critical point was 37.23 mmHg and 5.31 mmol/L. Conclusion TcPO₂ is decreased and ox-LDL is increased in patients with restenosis after interventional therapy for LASO, both indicators have certain predictive value for restenosis, and combined detection of the two can reflect restenosis condition more completely.

Abstract:Adipose tissue fibrosis is caused by improper remodeling of the extracellular matrix, which leads to abnormal deposition in adipose tissue. As the hallmark of adipose tissue dysfunction, adipose tissue fibrosis has a close relationship with metabolic dysfunction such as obesity and insulin resistance. Therefore, reversing adipose tissue fibrosis can restore the function of adipose tissue, enhance the sensibility of insulin and then improve patients' metabolic abnormalities such as obesity. This review summarizes the latest research progress in the pathological mechanism of adipose tissue fibrosis and the potential drug targets for reversing adipose tissue fibrosis, in order to provide a greater insight into this pathological process and provide the direction of drug development.

Abstract:Ferroptosis is a newly discovered iron-dependent mode of programmed cell death, characterized by intracellular iron overload, glutathione depletion and lipid peroxidation. Hypertension is one of the important risk factors for the occurrence of cardiovascular and cerebrovascular diseases, and more and more studies have shown that hypertension is closely related to ferroptosis. This article will review the association between ferroptosis and hypertension, as well as the possible mechanisms of ferroptosis affecting hypertension, in order to provide new ideas for the treatment of hypertension.

Abstract:Atherosclerosis (As) is a lipid-driven chronic inflammatory vascular disease, the pathogenesis of which primarily includes lipid metabolism disorders and inflammatory responses. Calpain, a calcium-dependent cysteine protease, has been shown to be involved in the development of As. Especially calpain in macrophages plays an important role in regulating lipid metabolism and inflammatory responses in As. This article reviews the possible pathways and molecular mechanisms of macrophages calpain involvement in As trying to provide new ideas for the prevention and treatment of As.

Abstract:Atherosclerosis (As), a complex chronic inflammatory disease, that is a major cause of cardiovascular diseases (CVD) including ischemic heart disease and stroke, causing high morbidity and mortality worldwide. In recent years, the role of gut microbiota in atherosclerosis has received extensive attention. However, as an important anti-atherosclerosis beneficial bacteria, Akkermansia is rarely reviewed. In-depth study of the pathogenesis and potential treatment of atherosclerosis is the focus of current medical development. Therefore, this article reviews how Akkermansia exerts its anti-atherosclerotic effect and its relationship with anti-atherosclerotic drugs.