内皮细胞缺氧后miR-210、miR-92a、miR-126表达及意义

目的进一步探讨缺氧后血管新生的调控机制。方法常规方法培养人微血管内皮细胞,低氧培养箱制备内皮细胞缺氧模型（观察组）,正常细胞作为对照组。采用real time PCR法检测两组内皮特异性microRNA（miR-210、miR-92a、miR-126）表达变化。结果与对照组比较,miR-210、miR-92a、miR-126分别上调了（5.19±0.99）、（3.02±0.88）、（3.87±0.83）倍,P均〈0.05。结论缺氧可促使内皮细胞特异性microRNA表达改变,此可能为缺氧后血管新生的主要调控机制。     

MicroRNA-15a/b are up-regulated in response to myocardial ischemia/reperfusion injury

Objective Several studies have indicated that miR-15a, miR-15b and miR-16 may be the important regulators of apoptosis. Since attenuate apoptosis could protect myocardium and reduce infarction size, the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion (I/R) injury. Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo, while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation (H/R). Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R. Results Compared to those of the controls, I/R or H/R induced apoptosis of cardiomyocytes was significantly increased both in vivo (24.4% ± 9.4% vs. 2.2% ± 1.9%, P < 0.01, n = 5) and in vitro (14.12% ± 0.92% vs. 2.22% ± 0.08%). The expression of miR-15a and miR-15b, but not miR-16, was increased in the mice I/R model, and the results were consistent in the H/R model. Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury, therefore, down-regulation of miR-15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.     

血清miR-21、miR-148b、miR-200a、miR-200b在原发性肝癌患者中的表达及临床诊断意义

目的研究原发性肝癌(HCC)患者血清中微小RNA(miR)-21、miR-148b、miR-200a、miR-200b的表达及临床诊断意义。方法应用实时荧光定量逆转录-聚合酶链反应(qRT-PCR)检测西南医科大学附属医院收治的93例HCC患者(病例组)、48例肝脏良性病变(良性组)、48例慢性肝病患者(慢性肝病组)和50例健康体检者(对照组)血清中miR-21、miR-148b、miR-200a、miR-200b的表达。统计学分析各指标与临床病理特征之间的关系,受试者工作特征(ROC)曲线分析各指标的诊断价值。结果病例组血清miR-21与miR-200b表达显著高于良性组、慢性肝病组及对照组(均P<0.05),miR-200a与miR-148b表达显著低于良性组、慢性肝病组及对照组(均P<0.05)。HCC患者血清miR-21与miR-200b表达与病理分期、病理分级及肿瘤直径有关(均P<0.05),与性别、年龄、有无肝硬化及乙型肝炎病毒(HBV)抗原是否阳性无关(均P>0.05)。血清miR-200a与miR-148b表达与病理分期、病理分级有关(均P<0.05),与性别、年龄、肿瘤直径、有无肝硬化及HBV抗原是否阳性无关(均P>0.05)。miR-21、miR-148b、miR-200a、miR-200b及四项指标联合的ROC曲线下面积(AUC)分别为0.821、0.668、0.772、0.734、0.922。联合诊断的敏感性和特异性均高于任一单一指标。结论HCC患者血清miR-21与miR-200b表达上调,而miR-200a与miR-148b表达下调。并且其表达与病理分期、病理分级有关,联合检测血清miR-21、miR-148b、miR-200a、miR-200b有希望成为新的诊断HCC的肿瘤标志物。     

ADAMTS-4 and -8 are inflammatory regulated enzymes expressed in macrophage-rich areas of human atherosclerotic plaques

ObjectivesRemodeling of extracellular matrix (ECM) plays an important role in inflammatory disorders such as atherosclerosis. ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) is a recently described family of proteinases that is able to degrade the ECM proteins aggrecan and versican expressed in blood vessels. The purpose of the present study was to analyze the expression and regulation of several ADAMTSs before and after macrophage differentiation and after stimulation with IFN-γ, IL-1β and TNF-α. ADAMTS expression was also examined during atherosclerosis development in mice and in human atherosclerotic plaques.Methods and resultsReal time RTPCR showed that, of the nine different ADAMTS members examined, only ADAMTS-4 and -8 were induced during monocyte to macrophage differentiation, which was also seen at protein level. Macrophage expression of ADAMTS-4, -7, -8 and -9 mRNA were enhanced upon stimulation with IFN-γ or TNF-α. Furthermore, immunohistochemical analyses revealed that ADAMTS-4 and -8 were expressed in macrophage rich areas of human atherosclerotic carotid plaques and coronary unstable plaques. In addition, ADAMTS-4 expression was upregulated during the development of atherosclerosis in LDLR^?/?ApoB^100/100 mice. Whereas ADAMTS-4 expression was low in non-atherosclerotic aortas, it was significantly higher in aortas from 30–40-week old atherosclerotic animals.ConclusionThe present study suggests that ADAMTS-4 and -8 are inflammatory regulated enzymes expressed in macrophage-rich areas of atherosclerotic plaques. This is the first study associating ADAMTS-4 and -8 expression with atherosclerosis. However, further experiments are required to understand the physiological and pathological functions of ADAMTS in the vascular wall, and tools to measure ADAMTS activity need to be developed.     

Association study of miR-146a,miR-149, miR-196a2, and miR-499 polymorphisms with venous thromboembolism in a Korean population

Journal of Thrombosis and Thrombolysis - Despite much progress in microRNA (miRNA) research, information regarding the association between miRNAs and venous thromboembolism (VTE), especially in...     

Regulatory effects of miR-146a/b on the function of endothelial progenitor cells in acute ischemic stroke in mice

The study aims to explore how microRNA-146a/b (miR-146a/b) regulates the function of endothelial progenitor cells (EPCs) in acute ischemic stroke in mice. Eighty male SPF C57BL/6J mice were evenly divided into the model-6 h, model-12 h, model-24 h (mice suffered from middle cerebral artery occlusion [MCAO] for 6 h, 12 h and model-24 h) and normal groups. EPCs were transfected and assigned into the control, MCAO, MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. The qRT-PCR was used to detect miR-146a/b expression in EPCs. Expressions of tumor necrosis factor receptor-associated factor 6 (TRAF6) and interleukin-1 receptor-associated kinase 1 (IRAK1) were detected using western blotting. Cell proliferation and migration of EPCs were testified using CCK-8 assay and scratch test, respectively. Angiogenesis ability of EPCs was observed under microscope. MiR-146a and miR-146b expressions were lower in the model groups than the normal group. There were up-regulated TRAF6 and IRAK1 expressions in the model-6 h, model-12 h and model-24 h groups compared with the normal group. And there were down-regulated TRAF6 and IRAK1 expressions in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups than in the MCAO group. Compared with the control group, the proliferation, migration and angiogenesis ability of EPCs were significantly lower in the MCAO group, but higher in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. Besides, the miR-146a/b group showed more enhancement than the MCAO-miR-146a and MCAO-miR-146b groups. MiR-146a/b could down-regulate the TRAF6 and IRAK1 expressions and promote proliferation, migration and angiogenesis ability of EPCs, which was important for recovery of patients with hyperacute ischemic stroke.     

Endoglin, a TGF-beta receptor-associated protein, is expressed by smooth muscle cells in human atherosclerotic plaques

Endoglin is a transmembrane protein that is found in association with transforming growth factor-beta (TGF-beta) superfamily receptor complexes and has an expression pattern that appears to be restricted primarily to endothelial cells, activated macrophages, trophoblasts, and fibroblasts. Since mutations in endoglin have been shown to be linked to hereditary hemorrhagic telangiectasia type 1, a disease manifested as vascular malformations characterized by excessive layers of vascular smooth muscle cells (VSMC), the expression of endoglin was investigated in VSMC. In vivo, the majority of SMC in human atherosclerotic plaques expressed high levels of endoglin, while endoglin was not detected in SMC from samples of the normal arterial wall. In vitro studies demonstrate that human aortic smooth muscle cells (HASMC) express the L-isoform of endoglin. Like endothelial cells, HASMC express endoglin protein as a dimer on the cell surface that binds TGF-beta1. In vitro, endoglin expression by HASMC is upregulated in response to TGF-beta1, suggesting that the presence of this factor in the atherosclerotic plaque might be responsible for the increased expression of endoglin. The demonstration of increased levels of endoglin in VSMC in human atherosclerotic plaques suggests a role for SMC endoglin in the maintenance of vascular integrity and in the response of the vessel wall to injury.     

Study on Dangua recipe regulating miR-34a/Nampt Axis in diabetic rats

<正>Objective To study the effect of Dangua Recipe regulating microRNA(miR)-34a/Nampt niacin nicotinate phosphoribosyl transferase(Nampt) axis controlling glycolipid metabolism.Methods After being raised for4 weeks with high-fat and high-sugar chow,the diabetic model was established by intraperitoneal injection of streptozotocin(STZ).According to the blood glucose levels,28 diabetic model rats were selected and received high-fat and high-sugar diet continuously.     

Evaluation of SNPs in miR-196-a2, miR-27a and miR-146a as risk factors of colorectal cancer

AIM: To investigate whether selected single nucleotide polymorphisms (SNPs) in miR-196a2, miR-27a and miR-146a genes are associated with sporadic colorectal cancer (CRC).METHODS: In order to investigate the effect of these SNPs in CRC, we performed a case-control study of 197 cases of sporadic CRC and 212 cancer-free controls originating from the Central-European Caucasian population using TaqMan Real-Time polymerase chain reaction and allelic discrimination analysis.RESULTS: The genotype and allele frequencies of SNPs were compared between the cases and the controls. None of the performed analysis showed any statistically significant results.CONCLUSION: Our data suggest a lack of association between rs11614913, rs895819 and rs2910164 and colorectal cancer risk in the Central-European Caucasian population, a population with an extremely high incidence of sporadic colorectal cancer.     

Immunohistochemical localization of Betacellulin, a member of epidermal growth factor family, in atherosclerotic plaques of human aorta

Betacellulin (BTC), a new member of the EGF family, has been reported to be a potent mitogen for rat vascular smooth muscle cells (SMCs). BTC mRNA is known to be expressed in several human organs. However, the localization of BTC in human vascular tissues has not yet been clarified. We investigated whether or not BTC protein is involved in the pathogenesis of human atherosclerosis. Recombinant human BTC showed a mitogenic activity on cultured human aortic SMCs by measuring [3H]thymidine incorporation. The immunohistochemical localization of BTC, SMCs, macrophages, EGF receptors and ErbB4 was examined in autopsied human aortas. BTC was detected in both intimal and medial SMCs of the aortic wall. The percentage of BTC-positive medial SMCs in early types of atherosclerotic lesions decreased with age, but in adult, it was significantly higher in advanced types than in early types of atherosclerotic lesions. BTC-positive SMCs were predominantly localized in the medial side of the intima. Furthermore, numerous BTC-positive SMCs and macrophages were observed around the core lesion of atherosclerotic plaques. Receptors for BTC, EGF receptor and ErbB4, were expressed on SMCs, suggesting that BTC is associated with EGF receptor family-mediated signaling. BTC is produced in human aortic tissue and might play important roles in atherogenesis.     

Expression of miR-146a and miR-155 in the urinary sediment of systemic lupus erythematosus

We studied the levels of miR-146a and miR-155 in the urine sediment of SLE patients. The levels of miR-146a and miR-155 in the urine sediment of 40 SLE patients who were receiving calcitriol treatment and 13 healthy controls were determined with real-time quantitative polymerase chain reaction. The levels of urinary miR-146a and miR-155 in patients with SLE were significantly higher than that in healthy controls. Calcitriol treatment reduced the levels of urinary miR-155 in patients with SLE. The level of urinary miR-146a significantly correlated with estimated glomerular filtration rate (r = 0.242, P = 0.008). The level of urinary miR-155 significantly correlated with proteinuria (r = 0.407, P < 0.001) and systemic lupus erythematosus disease activity index (r = 0.278, P = 0.002). The level of urinary miR-146a reversely correlated with the urinary expression of TNF-α (r = −0.247, P = 0.012). Our results suggested that miR-146a and miR-155 might play important roles in the pathophysiology of SLE and the levels of urinary miR-146a and miR-155 could be used as potential markers for diagnosis, disease activity, and therapeutic response.