高血压Ⅱ期患者细胞因子诱生水平的研究

高血压是一种以平滑肌细胞增生为主要病变的疾病。近来的研究结果表明:高血压时,血管平滑肌细胞处于“合成状态”,细胞可以分裂、增殖,并有迁移性,此时血管平滑肌细胞内myc mRNA增加了14％～100％。汤健等也发现自发性高血压大鼠血管平滑肌细胞、心肌和肝脏中myc基因的转录水平明显高于wky大鼠。除此之外,sis基因表达亦高于wky大鼠,而fos基因的表达则高出wky大鼠5～8倍。由此可见,myc、fos、sis癌基因的激活可能引起平滑肌细胞增殖和肥厚,进而导致高血压发生。     

大鼠颈动脉再狭窄模型的建立及其病理机制的初步研究

目的:建立再狭窄动物模型,探讨再狭窄的发生规律及病理机制.方法:模拟临床经皮冠状动脉成形术(Percutaneous Transluminal Coronary Angioplasty,PTCA)过程,造成大鼠颈动脉扩张及血管内膜的损伤,在此基础上系统地观测术后不同时相点内膜、中膜的增生以及细胞增殖的动态变化规律,以及血管内皮修复情况.结果:新生内膜于术后28天增厚达高峰,增生的血管内膜细胞中以α-Atin染色细胞为主,提示血管内膜的增生大多来自中膜的平滑肌细胞.损伤动脉壁细胞增殖,中膜增厚在14时达到高峰与术后即刻比较有显著性差异(P＜0.05),但35天中膜面积与术后第一天比较无显著性差异(P＞0.05.血管回缩管腔明显变窄.血管外弹力板周径和内弹力板周径在手术后当天有明显增加,与对侧比较差异显著(P＜0.05).但随后开始出现收缩性重构并在第14-28天达到高峰.手术后35天与对侧比较无显著性差异(P＞0.05).血管内膜剥脱后血管内膜第14天即可以见到部分再内皮化,28天血管内膜基本完全再内皮化.结论:损伤动脉狭窄的主要原因为血管的收缩性重构和内膜过度增生.     

血管平滑肌细胞在小鼠动脉血管发育与疾病中的作用研究进展

平滑肌细胞招募和分化成熟是胚胎动脉发育的重要步骤。分化的平滑肌细胞在成年动脉中处于静息状态,高表达与收缩有关的蛋白。平滑肌细胞的分化是可逆的,在血管损伤后发生去分化,静息状态转变为增殖状态。平滑肌细胞的不受控增殖造成血管狭窄甚至堵塞,这是堵塞性血管病的主要病理过程。血管损伤后修复过程在多个方面与血管胚胎发育过程相同。过去几年对在动脉发育和疾病中平滑肌细胞的作用研究有了新的进展。该文首先明确了血管平滑肌细胞具有异质性,不同血管的平滑肌细胞起源不同;其次,动脉胚胎发育中存在平滑肌自内而外逐层次序成熟的现象。最近研究还发现,动脉损伤修复后增生的平滑肌细胞来源具有单(或寡)克隆性。这些最新研究有助于更好地理解平滑肌细胞在动脉血管发育与疾病中的作用,为干预堵塞性血管病带来新的希望。     

血管平滑肌细胞收缩的分子机制研究进展

血管中的平滑肌细胞位于中膜,具有维持血管形态和保持血管张力的重要作用。在正常情况下,血管平滑肌细胞处于一种收缩表型,而当其受到生物化学物质、机械刺激作用后会转变成分泌表型,表现为收缩力下降,迁移、增殖能力增强以及分泌细胞外基质能力增强。这些异常变化会促进血管再狭窄和动脉粥样硬化等疾病的发生与发展,因此研究其分子机制至关重要。本文主要概括论述参与调节血管平滑肌细胞收缩的分子机制研究进展。     

高压力对体外培养动脉中膜平滑肌细胞增殖及增殖相关蛋白和生长因子的影响

为了探讨力学因素在血管重建中的作用和机制,观察在单纯高压力条件下血管平滑肌细胞增殖及其相关蛋白和生长因子的变化。应用血管体外应力培养系统,在施加单纯压力的条件下培养猪颈总动脉。按压力大小,将培养的血管分为高压力(21．3kPa)组和正常压力(13．3kPa)组。两组血管均分别培养1、4和7d。免疫组织化学检测血管中膜平滑肌细胞的α-肌动蛋白,增殖细胞核抗原、血小板源性生长因子A、转化生长因子13l及P53蛋白的变化。结果显示：在高压力的作用下,随着培养时间的延长,α-肌动蛋白呈减少的趋势;增殖细胞核抗原,转化生长因子131、P53持续增多;血小板源性生长因子A先增加而后有所减少。说明高压力可明显促进血管平滑肌细胞表型的转变,发生增殖现象。提示高压力可能通过调节血小板源性生长因子A、转化生长因子131及P53蛋白的表达来调控血管平滑肌细胞的增殖。     

α-平滑肌肌动蛋白在人胸主动脉夹层中的表达

目的:探讨胸主动脉壁中α-平滑肌肌动蛋白(α-SMA)的表达与胸主动脉夹层(TAD)的关系。方法:采集人TAD的动脉壁组织和正常人胸主动脉壁组织,采用Western Blotting和免疫组织化学方法检测α-SMA在组织中的表达程度。结果:在DA组中,α-SMA的表达明显减少,血管平滑肌细胞(VSMC)以增殖表型为主。结论α-SMA的减少主要发生在血管中膜层的VSMC中,细胞发生表型变化,导致中膜弹性变差,发生夹层病变。因此,α-SMA可能在TAD的发病中具有重要作用,值得进一步深入探讨。     

对去血清后HITASY细胞分子表达及表型分析

以人血管平滑肌细胞克隆株HITASY为实验材料,探讨HITASY细胞分子表达与表型转换间的关系,为阐明血管新生内膜形成及再狭窄病理机制提供实验依据.实验表明,在含血清或去血清培养条件下,平滑肌细胞于体外发生表型转换.去血清后细胞外基质蛋白合成中止,增殖及移行能力趋于降低,细胞特异性标志物平滑肌α肌动蛋白、肌球蛋白重链及钙调结合蛋白等的表达随去血清时间延长而增加.进一步实验证实,在血管活性介质作用下,胞液钙离子浓度骤增产生膜信号级联反应,引发细胞面积减小而显示收缩功能.上述处于分化表型的细胞经补加血清后其表型特征又恢复到原有去分化型,提示体外培养人平滑肌细胞可发生表型转换.为验证去血清诱导表型转换过程中相关基因的表达变化,用差异显示PCR筛选出E1A激活基因阻遏子,在细胞处于分化表型时表达上调并对细胞增殖伴有较强抑制作用.     

慢性肺心病肺动脉平滑肌细胞的胶原合成：体内及体外观察

肺动脉构形重建（ｓｔｒｕｃｔｒｕｒａｌｒｅｍｏｄｅｌｉｎｇ）是慢性肺心病的重要血管病变,但其发生机制至今未完全明了。病变以血管中膜平滑肌细胞肥大、增生和细胞外基质（包括纤维性与非纤维性成分）增多导致的血管壁增厚、血管腔狭窄为特征。本文用天狼星红－偏振光显微镜观察,真彩色全自动图像分析法,测量１０例慢性肺心病尸检肺小动脉中膜厚度及中膜内Ⅰ、Ⅲ两型胶原的含量和所占的百分比。用３Ｈ－胸腺嘧院核苷和３Ｈ－脯氨酸掺入法,观察缺氧内皮细胞条件培养液（ＨＥＣＣＭ）对培养的肺动脉平滑肌细胞（ＰＡＳＭＣＳ）ＤＮＡ及胶原合成的影响。结果：（１）肺心病组４３７支肌型肺动脉平均中膜厚占血管直径的百分值高于对照组５±１．０８％;（２）肺心病组的Ⅰ型和Ⅲ型胶原面积分别占中膜面积的５４．６２％和５１９％,而对照组两型胶原占中膜面积小于２％。（３）ＨＥＣＣＭ组平滑肌细胞的３Ｈ－ＴｄＲ和３Ｈ－脯氨酸掺入量（ｃｐｍ值）,均明显高于常氧对照组（ＮＥＣＣＭ）,两组相比,有显著性差异（Ｐ＜０．０１）。（４）细胞周期分析,ＨＥＣＣＭ组平滑肌细胞的Ｇ０／Ｇ１期细胞数百分值比ＮＥＣＣＭ组少２８％,Ｇ２＋Ｍ期细胞百分值则比ＮＥＣＣＭ组高３０％。可以认为,缺     

血管内皮生长因子基因对兔髂动脉内膜损伤的组织形态变化过程的影响

探讨血管内皮细胞的特异丝裂原－血管内皮生长因子（VEGF）基因阻止血管内膜损伤后形成再狭窄的组织变化过程。建立球囊拉伤血管内膜的兔髂动脉模型,将携带VEGF目的基因的真核表达载体pcDNA3/VEGF经多聚赖氨酸处理的PTCA球囊导管导入拉伤的血管内膜。VEGF基因组拉伤2周时血管内壁有VEGF mRNA和蛋白的高表达。血管内膜内皮化较快。2周时即有许多血管内皮细胞呈岛状分布。4周时内膜基本恢复光滑。内膜平滑肌细胞增生明显减少,而对照组2周时血管内膜粗糙,基底膜暴露,拉伤后4周仍无内皮细胞再生,最后形成虫蚀样改变。血管中膜平滑肌细胞穿过内弹性膜进入内膜并大量增生,内膜增厚。VEGF基因定位导入血管内壁后。VEGF mRNA和蛋白高表达且发挥其生物学效应,内皮细胞岛状增生,加快内膜内皮化,减轻内膜增厚。     

SM22α在血管平滑肌细胞中的作用

平滑肌22α(SM22α)是平滑肌细胞(VSMC)骨架相关蛋白,通过与肌动蛋白的作用参与VSMC骨架重构,是近年发现的一种VSMC分化标志物,其表达具有平滑肌组织特异性和细胞表型特异性.血管平滑肌细胞(VSMC)表型转化是动脉粥样硬化、高血压等血管重塑性疾病的共同病理生理过程.VSMC表型转化过程中平滑肌特异基因的表达变化和细胞骨架的重构是当前研究的热点问题之一.本文就SM22α的结构特征及其在VSMC中的作用机制进行综述.     

Phenotypic switching of vascular smooth muscle cells in atherosclerosis,hypertension, and aortic dissection

Vascular smooth muscle cells (VSMCs) play a critical role in regulating vasotone, and their phenotypic plasticity is a key contributor to the pathogenesis of various vascular diseases. Two main VSMC phenotypes have been well described: contractile and synthetic. Contractile VSMCs are typically found in the tunica media of the vessel wall, and are responsible for regulating vascular tone and diameter. Synthetic VSMCs, on the other hand, are typically found in the tunica intima and adventitia, and are involved in vascular repair and remodeling. Switching between contractile and synthetic phenotypes occurs in response to various insults and stimuli, such as injury or inflammation, and this allows VSMCs to adapt to changing environmental cues and regulate vascular tone, growth, and repair. Furthermore, VSMCs can also switch to osteoblast-like and chondrocyte-like cell phenotypes, which may contribute to vascular calcification and other pathological processes like the formation of atherosclerotic plaques. This provides discusses the mechanisms that regulate VSMC phenotypic switching and its role in the development of vascular diseases. A better understanding of these processes is essential for the development of effective diagnostic and therapeutic strategies.     

Small artery remodelling in diabetes

The aim of this article is to briefly review available data regarding changes in the structure of microvessels observed in patients with diabetes mellitus, and possible correction by effective treatment. The development of structural changes in the systemic vasculature is the end result of established hypertension. In essential hypertension, small arteries of smooth muscle cells are restructured around a smaller lumen and there is no net growth of the vascular wall, although in some secondary forms of hypertension, a hypertrophic remodelling may be detected. Moreover, in non-insulin-dependent diabetes mellitus a hypertrophic remodelling of subcutaneous small arteries is present. Indices of small resistance artery structure, such as the tunica media to internal lumen ratio, may have a strong prognostic significance in hypertensive and diabetic patients, over and above all other known cardiovascular risk factors. Therefore, regression of vascular alterations is an appealing goal of antihypertensive treatment. Different antihypertensive drugs seem to have different effect on vascular structure. In diabetic hypertensive patients, a significant regression of structural alterations of small resistance arteries with drugs blocking the renin–angiotensin system (angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers) was demonstrated. Alterations in the microcirculation represent a common pathological finding, and microangiopathy is one of the most important mechanisms involved in the development of organ damage as well as of clinical events in patients with diabetes mellitus. Renin–angiotensin system blockade seems to be effective in preventing/regressing alterations in microvascular structure.     

Renin-angiotensin system and vascular remodelling

The renin-angiotensin system (RAS) is compartmented between circulating blood and tissue pericellular space. Whereas renin and its substrate diffuse easily from one compartment to another, the angiotensin peptides act in the compartment where there are generated: blood or pericellular space. Renin is trapped in tissues by low and high affinity receptors. In the target cells, angiotensin II/AT1 receptor interaction generates different signals including an immediate functional calcium-dependent response, secondary hypertrophy and a late proinflammatory and procoagulant response. These late pathological effects are mediated by NADPH oxydase-generated free oxygen radicals and NFkappaB activation. In vivo, the tissue binding of renin and the induction of converting enzyme are the main determinants of the involvement of the RAS in vascular remodeling. The target cells of interstitial angiotensin II are mainly the vascular smooth muscle cells and fibroblasts, whereas the endothelial cells and circulating leukocytes are the main targets of circulating angiotensin II. In vivo, angiotensin II participates in the vascular wall hypertrophy associated with hypertension. In diabetes, as in other localized fibrotic cardiovascular diseases, the tissue effects of angiotensin II are mainly dependent on its ability to induce TGF-beta expression. In experimental atherosclerosis, angiotensin II infusion induces aneurysm formation mediated by activation of circulating leucocytes. In these models, the administration of angiotensin II antagonists has beneficial effects on pathological remodeling. Such beneficial effects of angiotensin II antagonists in localized pathological remodeling have not yet been demonstrated in humans.     

iPLA2β overexpression in smooth muscle exacerbates angiotensin II-induced hypertension and vascular remodeling

Objectives

Calcium independent group VIA phospholipase A₂ (iPLA₂β) is up-regulated in vascular smooth muscle cells in some diseases, but whether the up-regulated iPLA₂β affects vascular morphology and blood pressure is unknown. The current study addresses this question by evaluating the basal- and angiotensin II infusion-induced vascular remodeling and hypertension in smooth muscle specific iPLA₂β transgenic (iPLA₂β -Tg) mice.

Method and Results

Blood pressure was monitored by radiotelemetry and vascular remodeling was assessed by morphologic analysis. We found that the angiotensin II-induced increase in diastolic pressure was significantly higher in iPLA₂β-Tg than iPLA₂β-Wt mice, whereas, the basal blood pressure was not significantly different. The media thickness and media∶lumen ratio of the mesenteric arteries were significantly increased in angiotensin II-infused iPLA₂β-Tg mice. Analysis revealed no difference in vascular smooth muscle cell proliferation. In contrast, adenovirus-mediated iPLA₂β overexpression in cultured vascular smooth muscle cells promoted angiotensin II-induced [³H]-leucine incorporation, indicating enhanced hypertrophy. Moreover, angiotensin II infusion-induced c-Jun phosphorylation in vascular smooth muscle cells overexpressing iPLA2β to higher levels, which was abolished by inhibition of 12/15 lipoxygenase. In addition, we found that angiotensin II up-regulated the endogenous iPLA₂β protein in-vitro and in-vivo.

Conclusion

The present study reports that iPLA₂β up-regulation exacerbates angiotensin II-induced vascular smooth muscle cell hypertrophy, vascular remodeling and hypertension via the 12/15 lipoxygenase and c-Jun pathways.     

Mechanisms linking angiotensin II and atherogenesis

PURPOSE OF REVIEW: The concept that angiotensin II plays a central role in early atherogenesis, progression to atherosclerotic plaque, and the most serious clinical sequelae of coronary artery disease is the subject of considerable current interest. Results from recent large clinical trials confirm that blunting of the renin-angiotensin system through either angiotensin converting enzyme inhibition or angiotensin II type 1 receptor blockade incurs significant beneficial outcomes in patients with coronary artery disease. The exact mechanisms for these effects are not yet clear, but are suggested by studies demonstrating that suppression of the renin-angiotensin system is associated with muted vascular oxidative stress. RECENT FINDINGS: As most of the biological effects of the renin-angiotensin system occur through stimulation of the angiotensin II type 1 receptor, the focus of this review is on changes in the vascular wall mediated by this receptor and primarily related to endothelial and vascular smooth muscle cells, monocyte/macrophages and platelets. The interactions between angiotensin II and nitric oxide exert particular demands on the vascular capacity to adapt to dyslipidemia, hypertension, estrogen deficiency and diabetes mellitus that appear to exacerbate atherogenesis. Associated with each of these conditions is angiotensin II-mediated stimulation of macrophages, platelet aggregation, plasminogen activator inhibitor 1, endothelial dysfunction, vascular smooth muscle cell proliferation and migration, apoptosis, leukocyte recruitment, fibrogenesis and thrombosis. SUMMARY: Inhibition of the actions of angiotensin II serves a dual purpose: indirectly through reduction of mechanical stress on the vascular wall, and directly by diminished stimulation for vascular restructuring and remodeling. Collectively, data from studies published over the last year confirm and extend the notion that angiotensin II is a true cytokine prevalent at all stages of atherogenesis.     

Regulation of vascular smooth muscle cells and mesenchymal stem cells by mechanical strain

Vascular smooth muscle cells (SMCs) populate in the media of the blood vessel, and play an important role in the control of vasoactivity and the remodeling of the vessel wall. Blood vessels are constantly subjected to hemodynamic stresses, and the pulsatile nature of the blood flow results in a cyclic mechanical strain in the vessel walls. Accumulating evidence in the past two decades indicates that mechanical strain regulates vascular SMC phenotype, function and matrix remodeling. Bone marrow mesenchymal stem cell (MSC) is a potential cell source for vascular regeneration therapy, and may be used to generate SMCs to construct tissue-engineered vascular grafts for blood vessel replacements. In this review, we will focus on the effects of mechanical strain on SMCs and MSCs, e.g., cell phenotype, cell morphology, cytoskeleton organization, gene expression, signal transduction and receptor activation. We will compare the responses of SMCs and MSCs to equiaxial strain, uniaxial strain and mechanical strain in three-dimensional culture. Understanding the hemodynamic regulation of SMC and MSC functions will provide a basis for the development of new vascular therapies and for the construction of tissue-engineered vascular grafts.     

Mechanisms underlying mechanosensitivity of mesenteric afferent fibers to vascular flow

Spinal afferent neurons, with endings in the intestinal mesenteries, have been shown to respond to changes in vascular perfusion rates. The mechanisms underlying this sensitivity were investigated in an in vitro preparation of the mesenteric fan devoid of connections with the gut wall. Afferent discharge increased when vascular perfusion was stopped ("flow off"), a response localized to the terminal vessels just prior to where they entered the gut wall. The flow-off response was compared following pharmacological manipulations designed to determine direct mechanical activation from indirect mechanisms via the vascular endothelium or muscle. Under Ca(2+)-free conditions, responses to flow off were significantly augmented. In contrast, the myosin light chain kinase inhibitor wortmannin (1 microM, 20 min) did not affect the flow-off response despite blocking the vasoconstriction evoked by 10 microM l-phenylephrine. This ruled out active tension, generated by vascular smooth muscle, in the response to flow off. Passive changes caused by vessel collapse during flow off were speculated to affect sensory nerve terminals directly. The flow-off response was not affected by the N-, P-, and Q-type Ca(2+) channel blocker omega-conotoxin MVIIC (1 muM intra-arterially) or the P2X receptor/ion channel blocker PPADS (50 microM). However, ruthenium red (50 microM), a blocker of nonselective cation channels, greatly reduced the flow-off response and also abolished the vasodilator response to capsaicin. Our data support the concept that mesenteric afferents sense changes in vascular flow during flow off through direct mechanisms, possibly involving nonselective cation channels. Passive distortion in the fan, caused by changes in blood flow, may represent a natural stimulus for these afferents in vivo.     

Mineralocorticoid receptor: a critical player in vascular remodeling

Vascular remodeling is a pathological condition with structural changes of blood vessels. Both inside-out and outside-in hypothesis have been put forward to describe mechanisms of vascular remodeling. An integrated model of these two hypotheses emphasizes the importance of immune cells such as monocytes/macrophages, T cells, and dendritic cells. These immune cells are at the center stage to orchestrate cellular proliferation, migration, and interactions of themselves and other vascular cells including endothelial cells (ECs), vascular smooth muscle cells (VSMCs), and fibroblasts. These changes on vascular wall lead to inflammation and oxidative stress that are largely responsible for vascular remodeling. Mineralocorticoid receptor (MR) is a classic nuclear receptor. MR agonist promotes inflammation and oxidative stress and therefore exacerbates vascular remodeling. Conversely, MR antagonists have the opposite effects. MR has direct roles on vascular cells through non-genomic or genomic actions to modulate inflammation and oxidative stress. Recent studies using genetic mouse models have revealed that MR in myeloid cells, VSMCs and ECs all contribute to vascular remodeling. In conclusion, data in the past years have demonstrated that MR is a critical control point in modulating vascular remodeling. Studies will continue to provide evidence with more detailed mechanisms to support this notion.     

Monocyte chemoattractant protein-1 mediates angiotensin II-induced vascular smooth muscle cell proliferation via SAPK/JNK and ERK1/2

Abnormal vascular smooth muscle cells proliferation is the pathophysiological basis of cardiovascular diseases, such as hypertension, atherosclerosis, and restenosis after angioplasty. Angiotensin II can induce abnormal proliferation of vascular smooth muscle cells, but the molecular mechanisms of this process remain unclear. Here, we explored the role and molecular mechanism of monocyte chemotactic protein-1, which mediated angiotensin II-induced proliferation of rat aortic smooth muscle cells. 1,000 nM angiotensin II could stimulate rat aortic smooth muscle cells' proliferation by angiotensin II type 1 receptor (AT(1)R). Simultaneously, angiotensin II increased monocyte chemotactic protein-1 expression and secretion in a dose-and time-dependent manner through activation of its receptor AT(1)R. Then, monocyte chemotactic protein-1 contributed to angiotensin II-induced cells proliferation by CCR2. Furthermore, we found that intracellular ERK and JNK signaling molecules were implicated in angiotensin II-stimulated monocyte chemotactic protein-1 expression and proliferation mediated by monocyte chemotactic protein-1. These results contribute to a better understanding effect on angiotensin II-induced proliferation of rat smooth muscle cells.     

Haemodynamics and wall remodelling of a growing cerebral aneurysm: a computational model

We have developed a computational simulation model for investigating an often postulated hypothesis connected with aneurysm growth. This hypothesis involves a combination of two parallel and interconnected mechanisms: according to the first mechanism, an endothelium-originating and wall shear stress-driven apoptotic behavior of smooth muscle cells, leading to loss of vascular tone is believed to be important to the aneurysm behavior. Vascular tone refers to the degree of constriction experienced by a blood vessel relative to its maximally dilated state. All resistance and capacitance vessels under basal conditions exhibit some degree of smooth muscle contraction that determines the diameter, and hence tone, of the vessel. The second mechanism is connected to the arterial wall remodeling. Remodeling of the arterial wall under constant tension is a biomechanical process of rupture, degradation and reconstruction of the medial elastin and collagen fibers. In order to investigate these two mechanisms within a computationally tractable framework, we devise mechanical analogues that involve three-dimensional haemodynamics, yielding estimates of the wall shear stress and pressure fields and a quasi-steady approach for the apoptosis and remodeling of the wall. These analogues are guided by experimental information for the connection of stimuli to responses at a cellular level, properly averaged over volumes or surfaces. The model predicts aneurysm growth and can attribute specific roles to the two mechanisms involved: the smooth muscle cell-related loss of tone is important to the initiation of aneurysm growth, but cannot account alone for the formation of fully grown sacks; the fiber-related remodeling is pivotal for the latter.