Abstract:Atherosclerosis is a chronic inflammatory disease and an abnormal response of vascular walls to various injuries. Although there are many factors affecting atherosclerosis, the role of lymphatics in atherosclerosis has been neglected. Lymphatic vessels are the channels through which the interstitial fluid is returned to the blood circulation. Although a large number of lymphatic vessels have been found in atherosclerotic lesions, the relationship between the two is unclear. Recent studies have found that lymphatic vessels not only participate in the induction and solution of arterial inflammation, but also play an active role in cholesterol transport in atherosclerotic plaques. In addition, improving lymphatic function or promoting local induction of lymphangiogenesis may reduce the progression of atherosclerosis. Therefore, it is important to study the relationship between lymphatic vessels and development of atherosclerosis. This review article focuses on the relationship between lymphatic vessels and atherosclerosis, including inflammation, reverse cholesterol transport and immunity, in order to provide a new perspective for the study of intervention strategies for atherosclerosis.

Abstract:Aim To explore the role of β1 submit promoter methylation when hypertension and aging regulate the activity of large-conductance Ca²＋-activated K＋ channel in mesenteric arteries. Methods Male Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were studied at 3,6 months of age, blood pressure was noninvasively measured in the caudal artery, the mesenteric arteries were taken for the experiment. The whole-cell K＋ currents and BKCa single channel currents were measured using whole-cell and inside-out patch, respectively. Isometric contraction study was performed to investigate the contribution of BKCa channel to vascular tone regulation. Western blot was used to detect the expression of α and β1 submit of BKCa channel in mesenteric arteries. DNA bisulfite sequencing PCR was performed to investigate of the DNA methylation status at β1 gene (KCNMB1) promoter. Results At 3 months old, compared to WKY, the mean peak BKCa current density of SHR was significantly increased. Aging decreased the mean peak whole-cell current density of both SHR and WKY. The increases in the Po of BKCa channels that tamoxifen evoked in WKY were significantly lower than those of age-matched SHR. Meantime, hypertension increased the contribution of BKCa channels in vascular tone regulation. Compared with age-matched WKY, the protein expression level of the β1 subunit observed in SHR was always significantly increased. Aging reduced β1 subunit expression in mesenteric arteries. Hypertension decreased BKCa channel β1 submit promoter methylation in mesenteric arteries, in contrast, aging increased BKCa channel β1 submit promoter methylation in mesenteric arteries. Conclusions During hypertension, KCNMB1 gene demethylated and the expression of β1 submit increased, which enhanced the function of BKCa channel. Aging not only enhanced CpG methylation at KCNMB1 gene promoter but also upregulated β1 subunit expression, which diminished the function of BKCa channel.

Abstract:Aim To investigate the effect and mechanism of Erchen decoction and Taohong Siwu decoction containing serum on oxidized low density lipoprotein (ox-LDL) induced endothelial cell damage. Methods EA.hy926 cells were cultured in vitro, then randomly divided into control group, ox-LDL group, intervention group, inhibitor group.Cell apoptosis was detected by flow cytometry, malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) activity were detected by colorimetry, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in cell supernatant were detected by ELISA, and phosphatidylinositol 3-kinase (PI3K), phosphorylated AKT (p-AKT) and endothelial nitric oxide synthase (eNOS) protein expressions were detected by Western blot. Results Compared with control group, the level of TNF-α and IL-6 increased significantly (P＜0.01), MDA content in cells increased significantly (P＜0.01), SOD activity decreased (P＜0.05), NO level in supernatant decreased significantly (P＜0.01), content of reactive oxygen species (ROS) and apoptosis rate increased significantly, the protein expression level of PI3K, eNOS and the protein phosphorylation of AKT were significantly decreased (P＜0.01) in ox-LDL group. Compared with ox-LDL group, the level of TNF-α and IL-6 in cell supernatant decreased significantly (P＜0.01), MDA content in cells decreased significantly (P＜0.01), SOD activity increased significantly (P＜0.05), NO level in the supernatant increased significantly (P＜0.01), ROS content and apoptosis rate decreased significantly, the protein expression level of PI3K, eNOS and the protein phosphorylation of AKT were significantly increased (P＜0.01) in intervention group. Compared with intervention group, the level of TNF-α and IL-6 increased significantly (P＜0.01), MDA content in cells increased significantly (P＜0.01), SOD activity decreased (P＜0.05), NO level in supernatant decreased significantly (P＜0.05), content of ROS and apoptosis rate increased significantly, the protein expression level of PI3K, eNOS and the protein phosphorylation of AKT were significantly decreased (P＜0.01) in inhibitor group. Conclusion The serum containing of Erchen decoction and Taohong Siwu decoction can effectively protect ox-LDL induced endothelial cell injury, which may be related to activation of PI3K/AKT/eNOS signaling pathway.

Abstract:Aim To observe the effects of Kindlin-2 RNA interference on intimal hyperplasia in the balloon injured rat carotid arteries. Methods Kindlin-2 siRNA and negative control lentiviral vectors were constructed and produced, then balloon injured rat carotid arteries were infected with lentivirus. Carotid arteries were collected at 1 and 4 weeks after operation. The expression level of Kindlin-2 mRNA in the rat carotid arteries was detected by real-time quantitative PCR technique. The degree of intimal hyperplasia 4 weeks after balloon injury was assessed by HE staining. The immunofluorescence and histochemistry were used to detect the protein expression of Kindlin-2, α-actin, PCNA and β1-integrin in the carotid arteries. The synthesis of collagen fibers in the carotid arteries was investigated by Masson staining. Results Kindlin-2 RNA interference can significantly reduce the Kindlin-2 mRNA expression level in the rat carotid arteries at 1 week(P<0.05), and significantly inhibited intimal hyperplasia of carotid arteries 4 weeks after balloon injury (P<0.05). Compared with the negative control group, the protein expression levels of Kindlin-2, α-actin, PCNA and β1-integrin were significantly decreased in the carotid arteries of the Kindlin-2 group 4 weeks after operation. The synthesis of collagen fibers in the carotid arteries intima and media was also reduced. Conclusion Kindlin-2 RNA interference may inhibit vascular smooth muscle cell proliferation, migration and extracellular matrix synthesis to alleviate intimal hyperplasia.

Abstract:Aim In this experiment, human umbilical vein endothelial cells were induced by HHcy in vitro, and the inflammatory damage model of vascular endothelial cells was established, the effect of Diplacone on human umbilical vein endothelial cells was observed by pretreatment with Diplacone, and further explored the possible protective mechanism of Diplacone on vascular endothelial cells. Methods Human umbilical vein endothelial cells were used as the research object and were preincubated by Diplacone with different concentrations for 2 h, then 2 mmol/L Hcy was added to continue incubating for 12 h, changes of cell apoptosis rate were detected by flow cytometry; the level of oxidative stress was detected by ROS cell rate, MDA, GSH-Px,SOD in cells; protein expression and mRNA levels of NF-κB were detected by Western blot. Results Compared with control group, cell apoptosis rates were increased, oxidative stress indexes including ROS cell rate and MDA were increased and the activities of GSH-Px and SOD were decreased; the expression of NF-κB protein and mRNA increased significantly. After human umbilical vein endothelial cells were preincubated by Diplacone with different concentrations, compared with Hcy model group, cell apoptosis rates were decreased in 0.1～10 μmol/L; oxidative stress indexes including ROS cell rate and MDA were decreased and the activities of GSH-Px and SOD were increased; the expression of NF-κB protein and mRNA decreased significantly in 0.1～10 μmol/L Diplacone. Conclusion Diplacone can inhibit Hcy-induced endothelial damage in human umbilical vein endothelial cells and the mechanism may be through reducing oxidative stress and inhibiting the activity of NF-κB signaling pathway to reduce cell apoptosis rate and inhibit cell apoptosis.

Abstract:Aim To investigate the effect of peroxisome proliferator-activated receptor α (PPARα) agonist oleoylethanolamide (OEA) on the adhesion of monocytes to endothelial cells. Methods Western blot was used to detect the effects of OEA on the expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 in lipopolysaccharide-induced THP-1 cells and the expressions of vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells induced by tumor necrosis factor α (TNF-α). The adhesion of THP-1 cells and the expression of VCAM-1 protein were detected by TNF-α-induced monocyte-endothelial cell adhesion model after PPARα blocker MK886 blocked PPARα signaling pathway. Results 40 μmol/L OEA significantly inhibited MMP-2, MMP-9, VCAM-1 and ICAM-1 protein expression. OEA had a direct inhibitory effect on the monocyte-endothelial cell adhesion induced by TNF-α. MK886 partially reversed the inhibitory effect of OEA on monocyte-endothelial cell adhesion and VCAM-1 protein expression. Conclusion OEA can inhibit the adhesion of monocytes to endothelial cells, and its mechanism may be related to PPARα pathway.

Abstract:Aim To study the changes of gut microbiota of hyperlipemia guinea pigs by microbial 16S rDNA sequencing. Methods Guinea pigs were randomly divided into normal group and model group, with 10 in each group. Guinea pig models of hyperlipidemia were established by high fat diet. After feeding for 8 weeks, serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDLC) and high density lipoprotein cholesterol (HDLC) levels were detected. HE staining was performed to detect the histological changes of liver, aorta and colon. The changes of gut microbiota were detected by 16S rDNA analysis. Results The levels of TC, TG, LDLC and HDLC of model group were higher than that of control group (P<0.01). According to the histomorphological test, all guinea pigs in the model group showed severe fatty liver, but only one of them appeared early symptoms of atherosclerosis. The gut microbiota 16S rDNA high-throughput sequencing analysis result showed that, compared with control group, at the phyla level, the proportion of Synergistetes was obviously increased (P<0.05); at the genus level, the proportions of lachnospiraceae NK4A136, ruminococcus, helicobacter, odoribacter, allobaculum and caldicoprobacter of the model group were significantly reduced (P<0.05); the proportions of fretibacterium, lachnospiraceae XPB1014, pyramidobacter and enterorhabdus were obviously increased (P<0.05). Conclusions The compositions and proportions of gut microbiota in guinea pigs were significantly changed by high-fat feeding. The results of the experiment laid a theoretical foundation for the study of the mechanism of hyperlipidemia based on intestinal flora.

Abstract:Aim To evaluate the value of soluble suppression of tumorigenicity 2 (sST2) and interleukin 33 (IL-33) in cardiovascular impairment in children with Kawasaki's disease (KD). Methods A total of 282 children with KD admitted to the Department of Pediatrics of Hunan Provincial People's Hospital from December 2013 to January 2017 were selected, including cardiovascular involvement group (n＝35) and cardiovascular uninvolved group (n＝247), and 50 healthy children were as control group. The levels of plasma sST2 and IL-33 were detected in 3 groups of children.Changes and correlations of sST2, IL-33 and conventional inflammatory markers C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and white blood cell count (WBC) were analyzed in KD children with coronary artery lesion. Statistical methods were used to analyze data. Results The levels of plasma sST2 and IL-33 in the cardiovascular involvement group were significantly higher than those in the cardiovascular uninvolved group and the control group (P＜0.05). CRP, ESR and WBC in cardiovascular involvement group and cardiovascular uninvolved group were significantly higher than those in control group (P＜0.05). However, there were no significant differences in CRP, ESR and WBC between the cardiovascular involvement group and the cardiovascular uninvolved group (P＞0.05). After intravenous immunoglobulin therapy, the levels of sST2 and IL-33 were decreased significantly in cardiovascular involvement group (P＜0.05), while the levels of sST2 and IL-33 did not change significantly in cardiovascular uninvolved group (P＞0.05). Pearson correlation analysis showed that the levels of sST2 and IL-33 in KD children were positively correlated with the Z value of echocardiography. Logistic regression analysis showed that sST2 and IL-33 were independent risk factors for cardiovascular injury in KD children (P＜0.05). ROC curve analysis showed that sST2 and IL-33 had high diagnostic value for predicting cardiovascular injury in KD. Conclusions The levels of plasma sST2 and IL-33 are correlated with the degree of cardiovascular injury in children with KD. The analysis of sST2 and IL-33 is helpful to evaluate the degree of cardiovascular injury and predict the prognosis of children with KD.

Abstract:Aim To investigate the relationship between neutrophil/lymphocyte ratio (NLR) and severity of coronary artery stenosis and cardiovascular end point events after percutaneous coronary intervention (PCI) in patients with acute coronary syndrome (ACS). Methods A total of 302 patients admitted to our hospital with coronary angiography and PCI were enrolled in this study. All patients were divided into two groups according to Gensini scores:low score group (≤46.5 points) and high score group (＞46.5 points). The groups were compared according to their NLR and other parameters. All patients were divided into two groups according to the occurrence of cardiovascular end point events:event group and non-event group. The ROC curve was used to analyze the predictive value of NLR in cardiovascular end point events after PCI in patients with acute coronary syndrome. Results The prevalence of hypertension, lower high density lipoprotein cholesterol and NLR in the high score group were higher than those in the low score group (P＜0.05). Binary stepwise Logistic regression analysis showed that NLR and high density lipoprotein cholesterol were independent predictors of severity of coronary artery disease. The rates of elderly patients (≥60 years), hypertension, diabetes, lower high density lipoprotein cholesterol and NLR in the event group were higher than those in the non-event group (P＜0.05). The ROC curve analysis showed that the area under the curve of NLR in predicting cardiovascular end point events of PCI patients was 0.710 (95%CI was 0.632～0.788, P＜0.05). When the cut off value of NLR was 3.985, the diagnostic efficiency was the highest, with a sensitivity of 67.3% and a specificity of 70.4%. Kaplan-Meier curve analysis showed that the incidence of cardiovascular end point events were significantly higher in NLR＞3.985 group than in NLR＜3.985 group.Multivariate COX regression analysis showed that NLR, elderly, hypertension and diabetes were independent predictors of cardiovascular end point events after PCI. Conclusion The high NLR value may be an independent predictor of the severity of coronary artery stenosis and the incidence of cardiovascular end point events after PCI.

Abstract:Aim To observe the level of interleukin-17A (IL-17A) in patients with atrial fibrillation (AF), to assess its clinical value of thrombotic events in AF patients. Methods 67 controls (38 males and 29 females, average age 60.37±9.51 years), 65 patients with paroxysmal AF (35 males and 30 females, average age 58.92±8.59 years), and 88 patients with persistent AF (57 males and 31 females, average age 61.45±8.32 years) were enrolled in the present study, the expression of IL-17A was measured, then judged by creating the receiver operating characteristic curves (ROC). Serum levels of MMP-9, procollagen type Ⅲ and the left atrial ejection fraction (LAEF) were examined and compared. The patients with AF who were complicated by thrombotic events were followed-up, levels of IL-17A were further analyzed to assess the predictive value for thrombotic events in patients with AF. Results The level of serum IL-17A was increased in patients with AF. Area under the ROC of IL17A was 0.784 (P＜0.01), the sensitivity was 78.6%, the specificity was 69.1%. In the patients with AF who were complicated by thrombotic events, the level of serum IL-17A was significantly increased, while it was markedly decreased after regular anticoagulant therapy. The increased IL-17A level was significantly correlated with the levels of MMP-9, procollagen type Ⅲ and LAEF. Conclusion IL-17A might promote AF and thrombus event in AF patients.

Abstract:Aim To investigate the relationship between fragmented QRS wave (fQRS) of the classification of coronary collateral circulation and the improvement of heart function after percutaneous coronary intervention (PCI) in patients with chronic total occlusion. Methods 245 cases of occlusion patients were included from January 2013 to December 2016 in Shengjing Hospital of China Medical University, to statistically analyse the biochemical indexes, electrocardiogram, interventional data, Rentrop classification of collateral circulation, preoperative and postoperative heart function. Patients were divided into fragmented QRS group (fQRS group, n＝108) and no fragmentation of QRS group (nfQRS group, n＝137) according to the state QRS wave in electrocardiogram, to analyse the difference of fQRS wave, classification of collateral circulation, and the improvement of heart function in 6 month and 1 year after the operation in the two groups. Results The level of collateral circulation in patients of fQRS wave was higher than the nfQRS group, the ventricular ejection fraction and 6 minutes walk test distance were increased after PCI treatment in the two groups of patients, but the left ventricular end diastolic volume index and the brain natriuretic peptide were decreased, the above index change more significantly in fQRS group compared to nfQRS group, the difference was statistically significant, and the collateral circulation classification and the improvement of heart function were positively correlated. Conclusion In coronary chronic total occlusion patients, fQRS was positively related to classification of collateral circulation, percutaneous coronary interventional therapy can improve heart function in patients with chronic total occlusion, and the heart function in patients with fQRS were improved obviously.

Abstract:Aim To observe the effects of high-loading-dose trimetazidine on serum growth differentiation factor-15(GDF-15), monocyte chemoattractant protein-1(MCP-1), and high sensitive C-reactive protein(hs-CRP) level in patients with coronary heart disease underwent percutaneous coronary intervention(PCI). To evaluate the beneficial effects of high-loading-dose of trimetazidine before PCI in patients with coronary heart disease. Methods Totally 96 consecutive patients with coronary heart disease were divided into trimetazidine group(56 cases) and control group(40 cases). Standard medical therapies were given in all patients. In trimetazidine group, patients were given high-loading-dose of trimetazidine(60 mg) 30 minutes before PCI and maintenance dose at least 1 month after PCI. The GDF-15, MCP-1, and hs-CRP were measured in all enrolled subjects 24-hour after PCI. The net adverse clinical events(NACE) and major adverse cardiovascular cerebral events(MACCE) during the follow-up period were observed. Results The serum GDF-15 levels in trimetazidine group were lower than control group (737.24±299.85 ng/L vs 1175.84±523.52 ng/L, P=0.002). The MCP-1 and hs-CRP levels between the two groups had no statistical difference. Incidence of NACE in trimetazidine group were lower than control group(P=0.012), with statistical difference. Conclusions High-loading-dose of trimetazidine before PCI reduces GDF-15 level and incidence of NACE. GDF-15 is independent risk factor for prognosis of coronary heart disease.

Abstract:Aim To study the effect of atorvastatin combined with brisk walking on atherosclerotic plaques reversal. Methods 120 atherosclerotic patients were divided into statin group and combined group. Carotid atherosclerotic plaque area, carotid intima-media thickness and stable plaque rate were used to evaluate plaque reversal effect. Statin group was given atorvastatin calcium tablets 20 mg/d (Beijing Ale) orally. The combined group took a quick walk (4～5 days/week) on the basis of atorvastatin calcium tablets 20 mg/d. The follow-up was 2 years. Results The combined group was superior to statin group in reducing TG, TC, LDLC and increasing HDLC. The combined group was superior to statin group in carotid artery plaque IMT, maximum plaque area, plaque number and stable plaque rate. Conclusion The combination of statins and fast aerobic exercise can improve the anti-atherosclerotic effect and reduce the level and function of LDLC, TC and TG, increase the level and function of HDLC, increase the stability of plaque, reverse the plaque effect, and increase the anti-arteriosclerosis as the time prolongs.

Abstract:Aim To explore the correlation between plasma HDL function and coronary artery stenosis and plaque property in coronary artery disease (CAD) patients with normal HDLC levels (≥1.03 mmol/L). Methods 129 cases suspected CAD with normal HDLC levels in Yichang Hospital of Traditional Chinese Medicine from October 2015 to December 2017 were enrolled, and grouped according to the severity of coronary artery stenosis and plaque property by 64-slice spiral CT coronary angiography (CTA). Paraoxonase 1 (PON1) activity and HDL oxidation/antioxidant index were detected. Results Compared with non-CAD group, plasma HDLC level and PON1 activity were significantly decreased, and HDL oxidation/antioxidant index was increased in the CAD group, showing an oxidized state, while ApoAI level had no difference. Although there were no difference in HDLC and ApoAI levels, PON1 activity was gradually decreased and HDL oxidation/antioxidant index was increased, accompanying with increased stenosis of coronary artery. The data also showed that PON1 activity of calcified plaque group was higher than that of soft plaque group and mixed plaque group. There is a correlation between PON1 activity and HDLC, HDL oxidation/antioxidant index. Conclusion PON1 activity as an index of HDL function may be a potential biomarker for assessment of the plaque property and severity of CAD.

Abstract:Reverse cholesterol transport (RCT) is the only mechanism that clears cholesterol in the body and has positive significance for maintaining cholesterol homeostasis in vivo. MicroRNA (miRNA) is non-coding RNA that has the ability to transcriptionally regulate gene expression. Hundreds of miRNA have been identified in humans and they are involved in the regulation of almost all processes, including cholesterol transport, metabolism, and maintenance of cholesterol homeostasis. Due to their small size and the ability to specifically regulate gene expression, miRNA has gradually become targets for the regulation of dyslipidemia and other lipid related diseases. This article reviews the miRNA that mediate RCT, including miR-33, miR-19b, miR-144-3p, miR-223, and miR-378. The summary of how miRNA modulates cholesterol metabolism provides new ideas for the prevention and treatment of atherosclerosis.

Abstract:With the popularization of CT angiography, magnetic resonance angiography and digital subtraction angiography (DSA), the detection rate of intracranial artery stenosis is improved. Intracranial artery stenosis causes a diversity of clinical manifestation. For example, asymptomatic, dizziness and stroke and so on. As a multi-etiological disease, its pathogenesis is complex, and the recurrence rate of ischemic stroke is high. Intracranial artery stenosis have different distribution in age, sex and race. This may suggest that genetic factors play an important role. At present, some progress has been made in genetic factors of intracranial artery stenosis.

Abstract:Cells need cholesterol to survive, but excess cholesterol is toxic to cells, so cells need to regulate cholesterol homeostasis. Intracellular cholesterol is transported to the high density lipoprotein-apolipoprotein AI, which returns to the liver in reverse cholesterol transport (RCT) route, and ultimately excretes through feces. RCT is not only a physiological process necessary to maintain cellular cholesterol homeostasis, but also has potential atherosclerosis inhibition. Current studies have mainly focused on the initial pathway of cellular cholesterol efflux and the final metabolism of cholesterol in the blood, but little is known about how cholesterol can leave the blood. More and more studies have shown that HDL needs to be transported through lymphatic vessels during RCT to return to liver in which cholesterol is metabolized. Therefore, it is important to better understand the process of HDL flowing from blood to peripheral tissue, to change and promote the removal of intracellular cholesterol in the tissue space and how it is transported through lymphatic vessels. This review focuses on the relationship between lymphatic vessels and RCT, which provides a new strategy for the treatment of atherosclerotic vascular diseases.

Abstract:Homocysteine is a sulfur-containing amino acid needed by human body and an important intermediate metabolite of methionine cycle. In recent years, hyperhomocysteinemia as an independent risk factor of cardiovascular disease has attracted wide attention. At present, more and more evidence shows that hyperhomocysteinemia plays an important role in the occurrence and development of arrhythmia. The purpose of this review is to elucidate the relationship between hyperhomocysteinemia and arrhythmia, and to find new ways to prevent arrhythmia and reduce its mortality.

Abstract:Numerous randomized clinical trials have established statins as the major standard therapy for atherosclerotic diseases because these molecules decrease the plasma level of low density lipoprotein cholesterol and moderately increase the plasma level of high density lipoprotein cholesterol (HDLC). The reverse cholesterol transport (RCT) pathway, mediated by HDL particles, has a relevant antiatherogenic potential. An important approach to HDL-targeted therapy is not optimization of the HDLC level but by improving HDL function, enhancing plasma cholesterol clearance, and preventing and reducing atherosclerosis related inflammation. Small-molecule inhibitors of cholesteryl ester transfer protein increase the HDLC level in subjects with normal or low HDLC. LXR agonists can reduce As by increasing RCT. HDL therapies using reconstituted HDL are dramatically effective in animal models. And, in cell and animal models, studies have found that by interfering with certain gene targets, the cholesterol level and function of HDL can be improved. Our review of the literature leads us to conclude that HDL-targeted therapies have significant atheroprotective potential and thus may effectively treat patients with cardiovascular diseases.