Abstract:环状RNA(circRNA)是一类新型内源性非编码RNA,广泛存在于真核生物体中,因其含量丰富、序列保守、结构稳定等特性,是近年来热门的基因表达调节因子。越来越多的研究表明circRNA在影响内皮完整性、血管平滑肌细胞和免疫炎症细胞的功能,以及在驱动动脉粥样硬化(As)斑块起始和稳定中发挥重要作用,因此其可能作为As新型诊断生物标志物和治疗靶标。因此,文章就circRNA在As中研究现状,综述其在As中的功能以及调节机制。

Abstract:目的 研究环状RNA circRNA_005647抑制心肌肥厚相关基因表达的作用机制。方法 建立血管紧张素Ⅱ(AngⅡ)灌注诱导的心肌肥厚小鼠模型。原代分离获得C57BL/6乳小鼠心肌细胞(NMVC),AngⅡ处理NMVC建立心肌细胞肥大模型。利用NMVC,分别感染circRNA_005647重组腺病毒(rAd-circRNA_005647)和转染miR-99b-5p模拟物来研究对NMVC肥大的影响。通过双荧光素酶报告基因实验和RNA Pull-down实验验证circRNA_005647与miR-99b-5p的结合作用。实时荧光定量PCR和Western blot检测NMVC中心肌肥厚相关基因的mRNA和蛋白表达水平。结果 在AngⅡ诱导的小鼠心肌肥厚模型和心肌细胞肥大模型中,circRNA_005647及其宿主基因肌球蛋白IXA(Myo9a)表达升高。过表达circRNA_005647可抑制AngⅡ诱导的NMVC中肥厚相关基因心房钠尿肽(Anp)和肌球蛋白重链7(Myh7)基因表达升高。circRNA_005647与miR-99b-5p间存在结合作用。过表达circRNA_005647可抑制miR-99b-5p促心肌细胞肥大的作用。结论 circRNA_005647通过结合miR-99b-5p发挥抑制心肌细胞肥大的作用。

Abstract:目的 探讨miR-320对人脐静脉内皮细胞(HUVEC)内皮间质转化(EndMT)的影响及其调控机制。方法 用miR-320 mimics或miR-320 inhibitor处理HUVEC。Western blot检测内皮细胞标记物血小板内皮细胞黏附分子1(CD31)、血管内皮钙黏蛋白(VE-Cadherin)及间质细胞标记物α平滑肌肌动蛋白(α-SMA)、波形蛋白(Vimentin)的表达；划痕实验、Transwell实验检测miR-320对HUVEC迁移的影响；罗丹明鬼笔环肽染色检测miR-320对HUVEC细胞骨架的影响；噻唑蓝法检测细胞增殖；流式细胞仪检测细胞周期；Western blot检测miR-320对SRY相关高迁移率族蛋白4(SOX4)表达的影响。结果 miR-320 mimics上调HUVEC的VE-Cadherin、CD31表达,下调Vimentin、α-SMA表达；miR-320 mimics使HUVEC细胞骨架微丝变细,应力纤维减少,细胞迁移能力减弱,但增殖能力无明显改变。miR-320 inhibitor下调HUVEC的VE-Cadherin、CD31表达,上调Vimentin、α-SMA表达；miR-320 inhibitor使HUVEC微丝增粗,应力纤维增多,细胞迁移能力增强。miR-320 mimics下调SOX4的表达,miR-320 inhibitor上调SOX4的表达。结论 miR-320通过下调SOX4的表达抑制HUVEC的EndMT。

Abstract:目的 观察miR-181a-5p对动脉钙化细胞模型的影响,探讨其可能的作用机制。方法 培养人主动脉平滑肌细胞(HASMC),经成骨样分化诱导后使用茜红素染色检测细胞成骨样分化的程度,采用real-time PCR检测成骨样分化标志基因骨钙素(OCN)、Runt相关转录因子2(RUNX2)和碱性磷酸酶(ALP)的mRNA表达变化,以及miR-181a-5p的表达变化,分析miR-181a-5p与成骨样分化相关基因表达水平的相关性。miR-181a-5p mimic转染用于上调HASMC内miR-181a-5p的表达。使用TargetScan数据库预测miR-181a-5p发挥作用的靶基因。使用real-time PCR检测miR-181a-5p过表达后对靶基因表达的影响。结果 HASMC经诱导成骨样分化后,茜红素染色阳性区域和细胞内钙含量逐渐增加,细胞中成骨样分化相关基因OCN、RUNX2和ALP的表达随时间延长逐渐升高,而miR-181a-5p的表达逐渐下降(P＜0.05)。相关性分析显示miR-181a-5p的表达与OCN、RUNX2和ALP的表达均呈负相关(P＜0.05)。使用miR-181a-5p mimic转染HASMC后细胞内miR-181a-5p的表达高于对照mimic组,细胞内钙含量及RUNX2和OCN的表达均低于对照mimic组(P＜0.05)。TargetScan数据库预测结果显示miR-181a-5p可与促成骨样分化基因丝裂原活化蛋白激酶1(MAPK1)和丝裂原活化蛋白激酶8(MAPK8)的3′UTR区结合,miR-181a-5p mimic组HASMC中靶基因MAPK1和MAPK8的表达显著下调(P＜0.05)。结论 血管平滑肌细胞成骨样分化时下降的miR-181a-5p可抑制成骨样分化,其可能的机制是抑制促钙化信号转导分子MAPK1和MAPK8的表达。

Abstract:泡沫细胞的形成与积累是动脉粥样硬化的标志,研究发现靶向泡沫细胞的形成在动脉粥样硬化的治疗与预防中具有重要作用。对动脉粥样硬化相关微小RNA(miRNA)的研究表明,miRNA可直接靶向胆固醇代谢中相关蛋白的表达,调节泡沫细胞的形成。且近年来对循环miRNA的研究发现,部分循环miRNA有望成为抑制泡沫细胞形成的新靶点。文章综述了miRNA对泡沫细胞形成各阶段的影响,并讨论了循环miRNA在泡沫细胞形成中的潜在作用。

Abstract:长链非编码RNA H19(long non-coding RNA H19,H19)在腹主动脉瘤中过表达。在多种血管疾病模型中发现,H19调节多种微小RNA(如miR-let-7、miR-29、miR-21等)参与动脉硬化、平滑肌细胞凋亡和炎症反应等病理生理过程,与腹主动脉瘤的发生发展关系密切。文章就H19调节微小RNA影响腹主动脉瘤形成的潜在机制进行综述。

Abstract:Aim To investigate the effect of succinate on pyroptosis of human umbilical vein endothelial cells (HUVEC) and its regulatory mechanism. Methods HUVECs were treated with succinate analogue diethyl succinate (DS) for 24 h, and the content of succinate was detected by colorimetry. Western blot was used to detect the expressions of pyroptosis-related protein cysteinyl aspartate specific proteinase 1 (Caspase-1), interleukin-1β (IL-1β), IL-18, NOD-like receptor protein 3 (NLRP3), gasdermin D N termine (GSDMD-N). The effects of succinate on ATP and reactive oxygen species (ROS) production of HUVEC were detected by ATP assay kit and ROS fluorescent probe. ROS scavenger N-acetylcysteine (NAC) was used to observe the role of ROS in HUVEC pyroptosis induced by succinate. Dimethyl malonate (DMM), a succinate oxidation inhibitor, was used to detect the effect of succinate oxidative metabolism on ROS production. Results DS promoted the accumulation of succinate in HUVEC, up-regulated the expressions of pyroptosis-related proteins Caspase-1, IL-1β, IL-18, GSDMD-N and NLRP3, inhibited ATP production and up-regulated ROS production. NAC inhibited the production of ROS induced by succinate and down-regulated the expressions of above pyroptosis-related proteins. DMM down-regulated succinate-induced ROS production and HUVEC pyroptosis. Conclusion Succinate up-regulates ROS production through oxidative metabolism, thus promoting HUVEC pyroptosis.

Abstract:Aim To investigate the role of miR-21/PTEN/Akt signal pathway in resveratrol promoting angiogenesis of endothelial progenitor cells(EPC) isolated from rat bone marrow. Methods Mononuclear cells were isolated by density gradient centrifugation from rat bone marrow and then cultured with EGM-2MV containing 10% fetal bovine serum. 3～5 generations of EPC were used in the experiments. EPC were treated with resveratrol (20 μmol/L) for 12 h.And then matrigel, real-time PCR and Western blot were used to detect the angiogenesis, the expression of miR-21 and PTEN mRNA , and the PTEN protein and Akt phosphorylation level of EPC in vitro separately. The dual luciferase reporter system was employed to detect the binding of miR-21 and PTEN mRNA. Results Resveratrol (20 μmol/L) significantly promoted the angiogenesis of EPC in vitro (P＜0.05). In the meantime, resveratrol inhibited the expression of miR-21 which promoted the gene and protein expression of PTEN. PTEN furtherly inhibited the phosphorylation level of Akt. The results of the double luciferase reporter system showed that miR-21 bound to the 3′UTR of PTEN mRNA.Conclusion miR-21 regulated the angiogenesis ability of EPC promoted by resveratrol. The present study indicated that miR-21/PTEN/Akt signaling pathway may play an important role in EPC angiogenesis promoted by resveratrol in vitro.

Abstract:Aim The purpose of the present study was to investigate the potential role and mechanism of mitogen activated protein kinase kinase kinase kinase 4 (MAP4K4) in human umbilical vein endothelial cell (HUVEC) injury induced by oxidized low density lipoprotein (ox-LDL). Methods The cell injury model was induced by ox-LDL (100 mg/L). The mRNA expression of MAP4K4 was detected using RT-PCR assay. Western blot analysis was conducted to estimate the protein levels of MAP4K4, Bax, Bcl-2, C-Caspase-3, peroxisome proliferators-activated receptorγ (PPARγ) and ATP binding cassette transporter A1 (ABCA1). CCK8 assay was performed to measure cell viability. ELISA assay was applied to analyse cell apoptosis. The reactive oxygen species (ROS) production was measured by DCFH-DA assay.The content of malondialdehyde(MDA) and the activity of superoxide dismutase (SOD) and catalase (CAT)were detected by commercially available assay kits. Results The expression of MAP4K4 was elevated in ox-LDL-stimulated HUVEC. In addition, silencing of MAP4K4 apparently enhanced cell viability and attenuated HUVEC apoptosis induced by ox-LDL accompanied by a decrease in the expression of Bax and C-Caspase-3 and an increase in the expression of Bcl-2. Moreover, MAP4K4 knockdown remarkably alleviated the generation of ROS and the content of MDA accompanied with the enhanced activity of anti-oxidative enzyme system SOD and CAT triggerd by ox-LDL. Mechanically, ablation of MAP4K4 obviously activated PPARγ/ABCA1 signaling pathway. Conclusion Depletion of MAP4K4 relieves ox-LDL-induced vascular endothelial injury by reducing apoptosis and mitigating oxidative damage via PPARγ/ABCA1 activation, thus providing the basis for the therapeutic effect of MAP4K4 on the As.

Abstract:Aim To study the significance of T wave shape of aVR lead in predicting left ventricular function in patients with acute coronary syndrome (ACS) with three-vessel disease. Methods 160 patients with ACS three-vessel disease confirmed by coronary angiography were enrolled and divided into T-wave upright group (n＝72) and T-wave inversion group (n＝88) according to the T-wave morphology in lead aVR of electrocardiogram. The general data, left ventricular function, coronary artery lesion and clinical outcome were compared between the two groups. Results Ejection fraction (LVEF) was lower but proportion of LVEF ＜50%, serum N-terminal pro-brain natriuretic peptide (NT-proBNP), Gensini score, Syntax score and proportion of Syntax≥23 were higher in T-wave upright group than in T-wave inversion group(all P＜0.05). Compared with the T-wave inversion group, the upright group had more cases of using vasopressor and longer average hospitalization day (all P＜0.05). Spearman correlation analysis showed that upright T wave in lead aVR was negatively correlated with LVEF, and positively correlated with Gensini and Syntax score, NT-proBNP, LVEF and hospitalization day (all P＜0.05). Multivariate Logistic regression analysis showed that upright T wave in lead aVR was independently associated with ejection fraction. Conclusions The patients of upright T wave in lead aVR with ACS three-vessel disease had lower LVEF values and worse clinical outcome than that of T wave inversion. Upright T wave in lead aVR can be used to evaluate the left ventricular function in patients with ACS three-vessel disease.

Abstract:Aim To explore screening value of coronary calcification score combined with triglyceride/high density lipoprotein cholesterol (TG/HDLC) value for cardiovascular events in patients undergoing maintenance hemodialysis (MHD). Methods A total of 110 patients who underwent MHD were enrolled. They were divided into cardiovascular lesion group (n=40) and cardiovascular non-lesion group (n=70) according to presence or absence of cardiovascular events. The coronary calcification degree was scored by multi-slice spiral CT. The contents of serum TG and HDLC were measured. TG/HDLC value was calculated. The receiver operating characteristic curves (ROC curve) were applied to analyze screening value of coronary calcification score combined with TG/HDLC value for cardiovascular events in patients undergoing MHD. Results The body mass index (BMI), TG/HDLC and coronary calcification score in cardiovascular lesion group were significantly higher than those in cardiovascular non-lesion group, while HDLC level was significantly lower than that in cardiovascular non-lesion group (P＜0.05). TG/HDLC and coronary calcification score were independent risk factors for cardiovascular disease in patients undergoing MHD. The diagnostic efficiency of the combination detection was the highest, with the area under the curve (AUC) of 0.925 and 95%CI of 0.869～0.981, which were significantly better than those of single detection. Conclusion Coronary calcification score and TG/HDLC value are independent risk factors for cardiovascular disease in patients undergoing MHD, which can help screen out patients with cardiovascular disease risk from patients undergoing MHD.

Abstract:Aim To investigate the survival status and influencing factors of hospitalized patients with cerebral infarction complicated with carotid atherosclerosis for 1-year follow-up, so as to provide evidence-based basis for prolonging the survival period and reducing the mortality rate. Methods From April 2017 to August 8,8 inpatients with cerebral infarction and carotid atherosclerosis admitted to Changsha Central Hospital Affiliated to University of South China were enrolled and followed up for 1 year. According to their prognosis, the patients were divided into survival group and death group. The clinical characteristics of the two groups were compared. Cox proportional hazards regression model was used to analyze the factors affecting the survival status of the patients. Results Among 548 patients, 36 patients died during the follow-up period, with a mortality rate of 6.57%. Univariate analysis showed that marital status, education level, heart disease, age and triglyceride were the risk factors of death in patients with cerebral infarction and carotid atherosclerosis during the follow-up period (P＜0.05). Multivariate Cox proportional hazards regression model analysis showed that marital status (no spouse) (HR 1.6,5% CI 1.086-2.776), heart disease (HR 2.3,5% CI 1.156-4.752) increased the risk of death in hospitalized patients with cerebral infarction and carotid atherosclerosis, and high education level (HR 0.5,5% CI 0.044-0.952) reduced the death risk. Conclusions The hospitalized patients with cerebral infarction and carotid atherosclerosis with no spouse, low education level and heart disease have higher death risk. After discharge, the family members should strengthen the comprehensive care and humanistic care for the patients with cerebral infarction and carotid atherosclerosis who have no spouse or have heart disease, so as to reduce the risk of death.

Abstract:Aim To analyze the efficacy and safety of bioabsorbable polymer-coated drug-eluting stents in patients with coronary heart disease and diabetes mellitus. Methods The study searched database both at home and abroad, and retrieved the clinical randomized controlled trial (RCT) of comparison of bioabsorbable polymer-coated drug-eluting stent (BP-DES) and durable polymer-coated drug-eluting stent (DP-DES) in patients with coronary heart disease and diabetes mellitus. The efficacy and safety were evaluated by target lesion failure rate, target vessel revascularization rate, target lesion revascularization rate and cardiogenic mortality rate. Results A total of 14 RCT were included, of which 3 855 patients were treated with BP-DES and 2 916 patients with DP-DES, with a mean follow-up of 2.9 years. The overall target lesion failure rate of BP-DES versus DP-DES was 13.5% versus 12.8% (RR:1.7,5% CI:0.88～1.29, P=0.50). The overall cardiac mortality rate of BP-DES versus DP-DES was 6.2% versus 5.4% (RR:1.0,5%CI:0.90～1.34, P=0.35), and the overall target lesion and target vessel revascularization rates were not statistically different. There were no statistically significant differences in subgroup analysis according to follow-up time. Conclusion In patients with coronary heart disease and diabetes, BP-DES has a similar safety and efficacy profile to DP-DES.

Abstract:Early detection of arterial stiffness plays an important role in the prevention and treatment of cardiovascular diseases. Non-invasive detection methods can make early detection of arteriosclerosis important clinically. Pulse wave velocity is an indicator of non-invasive measurement of arterial stiffness and has been widely recognized by the medical community. The measurement of pulse wave velocity mainly includes the tonometry, oscillometry, ultrasound imaging, photoplethysmography, bioimpedance plethysmography, magnetic resonance imaging and the like. This paper reviews the research progress, advantages and disadvantages of these technologies and methods.

Abstract:Aim Myocardial fibrosis(MF)is characterized by extracellular matrix accumulation, fibroblast activation, and transformation into myofibroblast, which is one of the features of cardiac remodeling after cardiac injury. MF includes two basic types:reactive fibrosis and repair fibrosis. Two kinds of fibrosis often coexist in the process of ventricular remodeling. MF can lead to congestive heart failure, malignant arrhythmia and sudden death, and become an important cause of sustainable development and irreversible ventricular remodeling. Some studies have shown that metabolites of intestinal flora, including trimethylamine oxide, short chain fatty acids, indole oxyl sulfate and p cresol sulfate, are involved in the process of myocardial fibrosis and play an important role in the treatment of heart failure. It is expected to become a new target for the treatment of heart failure. In this paper, the mechanism of intestinal flora metabolites in myocardial fibrosis is described, and the research progress of improving myocardial fibrosis by intervention of intestinal flora is also introduced.