细胞外基质、基质水解酶与血管钙化

血管钙化常见于动脉粥样硬化、糖尿病、慢性肾功能衰竭及衰老的血管.近年来的研究证实血管钙化的发生是一种类似于生理性矿化的主动调节过程,而非单纯的钙磷的被动沉积.血管细胞外基质是血管的主要组成成分,对血管起支持、保护作用,且与血管壁细胞相互作用影响其粘附、增殖、迁移、分化等功能,同时又是各种生长因子和细胞因子的储存库.目前的研究显示,在血管钙化过程中细胞外基质的组成和表达可能发生了变化,并参与了对钙化进程的主动调节.基质水解酶可能通过基质降解依赖或非依赖的机制,在钙化的发生发展中起到重要作用.本文主要综述了在血管钙化过程中细胞外基质的变化及其对血管钙化的作用,以及基质水解酶对血管钙化过程可能的影响.     

焦磷酸盐与血管钙化

血管钙化是指促钙化因子与抑钙化因子失衡导致钙磷在血管壁的异常沉积,与心血管疾病的高发病率与高死亡率密切相关。细胞外焦磷酸盐作为内源性羟磷灰石结晶形成的抑制剂,在血管钙化的发生中发挥重要的作用。近年来的研究发现,焦磷酸盐代谢异常是导致某些家族遗传性疾病及慢性肾病患者发生血管钙化的重要机制。本文着重对焦磷酸盐在体内的代谢、焦磷酸盐及其代谢相关酶在血管钙化发生中的作用进行综述。     

血管中膜钙化的机制

血管钙化是一个主动的、可逆转的并且受到高度调控的病理过程,其特点是羟磷灰石结晶沉积于血管系统。血管钙化可分为血管内膜钙化和血管中膜钙化等。血管中膜钙化常见于糖尿病、慢性肾功能衰竭及衰老的血管中。本文主要综述了血管中膜钙化的相关机制。     

MiRNA在血管钙化调控中的作用

血管钙化(vascular calcification, VC)通常指机体钙磷代谢失衡导致磷酸钙盐在血管壁被动沉积而产生的一种病理过程。VC作为慢性肾脏疾病(chronic kidney diseases, CKD)患者的主要并发症之一,是CKD患者诱发心血管疾病(cardiovascular disease, CVD)导致高死亡率的重要因素。但是由于VC涉及的机制复杂,难以治愈,一旦发生难以逆转,目前临床上尚无有效的治疗方法。MiRNA是一种大小为21～23个碱基的单链小分子RNA,可以通过调控转录后基因的表达来影响细胞生物学功能。miRNA在VC过程中发挥了重要作用。本文对miRNA影响VC的相关机制进行了总结,为VC的临床治疗和诊断提供新的研究方向。     

外泌体调控血管钙化的研究进展

血管钙化是钙、磷等矿物质在血管壁的异常沉积,是一个主动的、活跃的、被高度调控的生物学过程,受到钙磷代谢紊乱、氧化应激、机械应力和炎症等多种因素的影响.作为胞间通讯的重要载体,外泌体已被证实与血管钙化的发生密切相关.一方面,外泌体可以通过在细胞间传递蛋白质、microRNAs等信息,促进血管平滑肌细胞发生成骨样表型转化以及矿物质沉积;另一方面,外泌体还可以诱导血管内皮细胞发生内皮-间充质转化,进而调控血管钙化的进程.但是,外泌体在内皮细胞和平滑肌细胞参与血管钙化进程中的具体作用及机制目前尚不完全清楚.本文就外泌体在调节平滑肌细胞的成骨样表型转化、矿物质沉积、microRNA转运以及内皮细胞的内皮-间充质转化中的作用进行了综述,以期为血管钙化的防治提供新思路.     

二甲双胍对华法令诱导大鼠动脉钙化的影响及其机制初探

目的:研究二甲双胍(metformin,MET)对华法令(Warfarin,WFN)诱导大鼠动脉钙化的影响及其机制。方法:将28只SD大鼠随机分为正常对照组、8周(W)钙化组、8W钙化+8W MET 100 mg/kg治疗组、8W钙化+8W MET 200 mg/kg治疗组。采用Von Kossa染色法检测胸主动脉组织中钙结节;邻甲酚肽络合酮比色法测定颈总动脉组织中钙沉积含量;免疫组化染色检测血管壁中成骨基因Runx2及血管平滑肌标志物α-SMA的表达;Western Blot检测血管壁中Runx2及自噬标志物LC3II的表达。结果:WFN干预8 W后,大鼠动脉中钙沉积含量显著增加(P0.01),MET治疗组主动脉钙含量与钙化组相比均显著降低(P0.01)。Von Kossa染色可见钙化组(WFN组)动脉壁中层黑色连续钙盐沉积条带,而进行MET治疗后,黑色条带明显减少,对照组未见黑色条带。免疫组化显示钙化组血管壁Runx2表达阳性,棕褐色染色较深,而α-SMA表达则显著下降,基本未见棕褐色染色沉积;MET治疗后能够逆转上述趋势。Western Blot显示钙化组血管壁Runx2表达明显上升,MET治疗后Runx2表达被抑制。此外,钙化过程中伴随着自噬标志物LC3II表达轻度上升;随着MET浓度升高,血管壁中自噬水平呈剂量依赖性显著升高。结论:二甲双胍能够有效抑制大鼠动脉钙化,减轻血管平滑肌细胞由收缩表型向成骨样表型转换,其机制可能与诱导自噬有关。     

动脉粥样硬化斑块钙化机制的研究进展

心血管疾病中动脉粥样硬化斑块的钙化是动脉粥样硬化的临床标志之一,主要发生在动脉血管的内膜.动脉粥样硬化斑块核心的钙化不会增加斑块的易损性,而粥样斑块纤维帽上的微钙化会加强纤维帽的周向应力,致使斑块的易损性增加.动脉粥样硬化斑块的钙化机制包括被动钙化和主动钙化,被动钙化受激素和局部信号的调节,主动钙化机制涉及复杂的细胞生命过程,基质囊泡、细胞凋亡、外泌体、氧化应激反应和细胞自噬等均参与了钙化过程.本文对动脉粥样硬化斑块的钙化机制的进展进行综述.     

衰老与血管钙化

血管钙化是指体内钙磷在血管壁的异常沉积,是一个主动的、高度可调节的、类似于骨形成的生物学过程。血管钙化是引起心血管疾病发病率、病死率升高的重要原因,也是危害中老年人身体健康的广泛存在的病理现象。近年研究表明,衰老与血管钙化存在密切关系,衰老可以通过诱导血管平滑肌细胞成骨样转化、内皮细胞释放囊泡、细胞外基质重塑、DNA损伤、炎症反应、磷代谢失衡以及抗衰老因子如Klotho和Sirtuin 1的表达减少而促进血管钙化。     

非编码RNA调控血管平滑肌细胞成骨样表型转化的研究进展

分化成熟的血管平滑肌主要功能是收缩血管、调节血管周径及血压等.在高磷、高糖、维生素D₃、炎症等因素的作用下,平滑肌细胞可转分化为成骨样细胞参与血管钙化的形成,诱发心脑血管不良事件.非编码RNA是经基因转录但不翻译为蛋白质的一类RNA总称,其通过调控多种细胞活动来参与机体的生理和病理过程.已有研究表明,非编码RNA可通过调控血管平滑肌细胞成骨样表型转化影响血管钙化的发生、发展.本文从微小RNA、长链非编码RNA、环状RNA几方面综述非编码RNA在血管平滑肌成骨样表型转化中的调节作用,有助于进一步了解血管钙化的分子机制以及发现防治血管钙化的新靶点.     

与钙化相关的骨基质蛋白研究的现状与展望

生物矿化（biomineralization）不仅包括正常情况下骨骼和牙齿的钙化也包括各种疾病时的钙化,如血管钙化、胎盘钙化,还包括珊瑚和贝类以及纳米细菌等不同物种的钙化。研究表明与骨骼钙化的相关的蛋白不仅发现于骨骼的钙化中,也同样发现于其他组织的钙化过程和其他物种的矿化过程中,因此本文就与骨骼钙化的相关蛋白做一综述,以期为进一步研究纳米细菌自身的矿化机制,探讨其引起的相关疾病的致病机制以及为进一步实验筛选合适的标志蛋白。     

Arterial calcifications

Arterial calcifications as found with various imaging techniques, like plain X-ray, computed tomography or ultrasound are associated with increased cardiovascular risk. The prevalence of arterial calcification increases with age and is stimulated by several common cardiovascular risk factors. In this review, the clinical importance of arterial calcification and the currently known proteins involved are discussed. Arterial calcification is the result of a complex interplay between stimulating (bone morphogenetic protein type 2 [BMP-2], RANKL) and inhibitory (matrix Gla protein, BMP-7, osteoprotegerin, fetuin-A, osteopontin) proteins. Vascular calcification is especially prevalent and related to adverse outcome in patients with renal insufficiency and diabetes mellitus. We address the special circumstances and mechanisms in these patient groups. Treatment and prevention of arterial calcification is possible by the use of specific drugs. However, it remains to be proven that reduction of vascular calcification in itself leads to a reduced cardiovascular risk.     

Mechanical programming of arterial smooth muscle cells in health and ageing

Arterial smooth muscle cells (ASMCs), the predominant cell type within the arterial wall, detect and respond to external mechanical forces. These forces can be derived from blood flow (i.e. pressure and stretch) or from the supporting extracellular matrix (i.e. stiffness and topography). The healthy arterial wall is elastic, allowing the artery to change shape in response to changes in blood pressure, a property known as arterial compliance. As we age, the mechanical forces applied to ASMCs change; blood pressure and arterial wall rigidity increase and result in a reduction in arterial compliance. These changes in mechanical environment enhance ASMC contractility and promote disease-associated changes in ASMC phenotype. For mechanical stimuli to programme ASMCs, forces must influence the cell’s load-bearing apparatus, the cytoskeleton. Comprised of an interconnected network of actin filaments, microtubules and intermediate filaments, each cytoskeletal component has distinct mechanical properties that enable ASMCs to respond to changes within the mechanical environment whilst maintaining cell integrity. In this review, we discuss how mechanically driven cytoskeletal reorganisation programmes ASMC function and phenotypic switching.     

Analysis of the effects of medicinal leech on arterial function in elderly volunteers by means of photoplethysmography with Angioscan-01

The state of the arterial wall in elderly patients after the application of medicinal leech was analyzed using an Angioscan-01 diagnostic device. The effect of one medicinal leech on the endothelial vasomotor function of small resistance arteries and medium-sized muscular arteries was estimated. Arterial wall rigidity was assessed by measuring the arterial stiffness index and the augmentation index during stress testing in the form of brachial artery occlusion. It has been shown that the application of one medicinal leech has not only local, but also system effect on arterial endothelium by improving its vasomotor function through normalization of arterial wall stiffness. This process is supposed to involve the salivary cell secretion of medicinal leech, which is able to raise the NO level both in cells and in extracellular fluid and to activate e-NOS and n-NOS, as it has been shown recently in the culture of human vascular endothelial cells (HUVEC).     

Factors influencing arterial wall mass transport

Arterial wall mass transport has particularly attracted attention because it may be implicated in the development of arterial disease, including arteriosclerosis. A short review is presented of the structure of the arterial wall and of studies of mass transport within it. Recent findings confirm that mass transport occurs across the entire arterial wall apparently from the lumen to the adventitial lymphatics. Evidence has emerged of inhomogeneity of the distribution volume for extracellular tracers in different layers of the wall. An attempt is made to interpret results which indicate that distension per se of arteries and increase of medial smooth muscle tone tend to compact the medial interstitium whereas pressure driven convection across the wall tends to expand that tissue. These findings imply a potentially important role of endothelial permeability, smooth muscle tone and luminal pressure in influencing solute transport in the wall and wall transport properties.     

Early calcification patterns of the iliac arteries and their relation to the arterial structure

Summary Gross calcifications of the common iliac and internal iliac arteries represent a common finding in newborn children and infants. In both arteries, the calcific deposits regularly appear in certain areas of the arterial luminal surface only, whereas the other parts of the arterial wall remain free of gross lesions even in cases with a pronounced calcification. In the common iliac artery, the lateral wall of the vessel and the adjacent sectors of the anterior and posterior wall represent the predilection site of calcific deposits. In the internal iliac artery, the gross calcifications have been regularly demonstrated in the dorso-medial wall. The predominant localisation of the calcification in these parts of the vessels and its absence in the others depend on the definite structural features of the arterial tube and different affinity for calcium of the individual structural elements. In both iliac arteries, only the primary internal elastic membrane undergoes early calcification. However, unlike the most muscular arteries, this membrane is not developed in the whole arterial circumference of the common iliac and internal iliac arteries, but is absent in large areas of their arterial luminal layer. In these areas, the subendothelial or subintimal elastic layers are formed by the networks of longitudinally arranged elastic fibers or membraneous elastic structures which arise from the elastic networks with the further growth. These elastic elements always stay free of calcific deposits. The structural features found in both iliac arteries may be important for the development of the later pathological changes.     

Calcification of human vascular smooth muscle cells: associations with osteoprotegerin expression and acceleration by high-dose insulin

Arterial medial calcifications occur often in diabetic individuals as part of the diabetic macroangiopathy. The pathogenesis is unknown, but the presence of calcifications predicts risk of cardiovascular events. We examined the effects of insulin on calcifying smooth muscle cells in vitro and measured the expression of the bone-related molecule osteoprotegerin (OPG). Human vascular smooth muscle cells (VSMCs) were grown from aorta from kidney donors. Induction of calcification was performed with beta-glycerophosphate. The influence of insulin (200 microU/ml or 1,000 microU/ml) on calcification was judged by measuring calcium content in the cell layer and by von Kossa staining. OPG was measured in the medium by ELISA. Histochemistry was used for determination of alkaline phosphatase (ALP). Bone sialoprotein (BSP) and OPG mRNA expressions were done by RT-PCR. beta-Glycerophosphate was able to induce calcification in human smooth muscle cells from a series of donors after variable time in culture. Decreased OPG amounts were observed from the cells during the accelerated calcification phase. High dose of insulin (1,000 microU/ml) accelerated the calcification, whereas lower concentrations (200 microU/ml) did not. Calcified cells expressed ALP and BSP activity in high levels. In conclusion, high concentration of insulin enhances in vitro-induced calcification in VSMCs. Altered OPG levels during the calcification raise the possibility that OPG may have a potent function in regulating the calcification process or it may represent a consequence of mineralization. Effects of insulin and modulations by OPG on the calcification process in arterial cells may play a role in the development of calcifications as part of the diabetic macroangiopathy.     

Interactions of arterial cells: III. Stathmokinetic analyses of smooth muscle cells cocultured with endothelial cells

Arterial endothelial cells (EC) or their conditioned medium (ECCM) can alter the proliferation of cocultured arterial smooth muscle cells (SMC). Previously, we have shown, as have others, that EC regulate the growth of cocultured SMC depending on the density of both cell types. To ascertain the rate of cell-cycle traverse in preconfluent arterial SMC cocultured with arterial EC or ECCM (derived from preconfluent EC), we have conducted a series of stathmokinetic experiments using flow cytometry to determine where specific changes may occur in the cell cycle. Results of our experiments indicate for the first time that ECCM stimulates the proliferation of preconfluent SMC by significantly shortening the residence times in the G1 and S phases of the cell cycle. The predominant relative effect occurs within the early G1 (G1A) compartment where pretreatment with ECCM shortens the residence time by approximately 55%. Furthermore, we have observed that preincubation of serum-free ECCM with antiplatelet-derived growth factor (PDGF) antibody abolishes any mitogenic effect on SMC. This suggests that EC secrete PDGF-like molecules which enhance the proliferation rate of preconfluent, cocultured SMC. These findings support the hypothesis that arterial EC may secrete mitogens which stimulate arterial SMC proliferation in the vascular wall.     

Impact of calcifications on patient-specific wall stress analysis of abdominal aortic aneurysms

As a degenerative and inflammatory desease of elderly patients, about 80% of abdominal aortic aneurysms (AAA) show considerable wall calcification. Effect of calcifications on computational wall stress analyses of AAAs has been rarely treated in literature so far. Calcifications are heterogeneously distributed, non-fibrous, stiff plaques which are most commonly found near the luminal surface in between the intima and the media layer of the vessel wall. In this study, we therefore investigate the influence of calcifications as separate AAA constituents on finite element simulation results. Thus, three AAAs are reconstructed with regard to intraluminal thrombus (ILT), calcifications and vessel wall. Each patient-specific AAA is simulated twice, once including all three AAA constituents and once neglecting calcifications as it is still common in literature. Parameters for constitutive modeling of calcifications are thereby taken from experiments performed by the authors, showing that calcifications exhibit an almost linear stress–strain behavior with a Young’s modulus E ≥ 40 MPa. Simulation results show that calcifications exhibit significant load-bearing effects and reduce stress in adjacent vessel wall. Average stress within the vessel wall is reduced by 9.7 to 59.2%. For two out of three AAAs, peak wall stress decreases when taking calcifications into consideration (8.9 and 28.9%). For one AAA, simulated peak wall stress increases by 5.5% due to stress peaks near calcification borders. However, such stress singularities due to sudden stiffness jumps are physiologically doubtful. It can further be observed that large calcifications are mostly situated in concavely shaped regions of the AAA wall. We deduce that AAA shape is influenced by existent calcifications, thus crucial errors occur if they are neglected in computational wall stress analyses. A general increase in rupture risk for calcified AAAs is doubted.     

Recanalization of occlusion of the peripheral segment of the femoral artery by the Kensey method]

The aim of the presented study was to analyse the surgical technique and results of recanalization of the peripheral segment of iliac artery with Kensey's technique. Altogether 16 patients were operated. Arterial patency was observed in 14 patients immediately after surgery. This number decreased to 11 after a 3-month follow-up. Complications in the form of perforation of the arterial wall, hemorrhage at the site of puncture and thrombotic lesions in the reconstructed arterial channel were seen in 4 cases. Authors' own experience suggests that Kensey's technique together with laser surgery are the treatment of choice in case of iliac arterial occlusion in patients, in whom classic surgery is contraindicated.     

Subset of cells immunopositive for neurokinin-1 receptor identified as arterial interstitial cells of Cajal in human large arteries

In the adventitia of large arteries, dendritic cells are located between nerve fibers, some of which contain substance P. The aim of the present study was to examine whether neurokinin 1 receptor (NK-1R) was expressed by dendritic cells in the arterial wall. Parallel sections of aortic and carotid artery segments were immunostained with anti-NK-1R and cell-type-specific antibodies. Dendritic cells in the arterial wall expressed NK-1R, albeit at a low level. Other cells, which intensely expressed NK-1R, were located along the border between the media and adventitia. They did not co-express any dendritic cell markers, including fascin, CD1a, S100, or Lag-antigen, and were negative for CD68, CD3, and mast cell tryptase. These NK-1R⁺ cells were laser-capture microdissected and studied by means of electron-microscopic analysis. The microdissected cells were in direct contact with nerve endings, and their ultrastructure was typical of the interstitial cells of Cajal present in the gastrointestinal tract. Further systematic electron-microscopic analysis revealed that the cells displaying the features typical of interstitial cells of Cajal were a basic element of the human arterial wall architectonics. Arterial interstitial cells of Cajal were negative for c-kit but they expressed vasoactive intestinal peptide receptor 1 (VIPR1). Destructive alterations of contacts between arterial interstitial cells of Cajal and nerve endings were observed in arterial segments with atherosclerotic lesions. The functional significance of the arterial interstitial cells of Cajal and their possible involvement in atherosclerosis and other vascular diseases need clarification.This work was supported by the St Vincents Clinic Foundation, Sydney, Australia.