血管平滑肌细胞凋亡在腹主动脉瘤发病中的作用

目的：探讨腹主动脉瘤（AAA）中膜血管平滑肌细胞（VSMC）密度降低的机制。方法：选取人体肾下AAA及正常腹主动脉组织（NA）标本,采用免疫组化及原位末端DNA标记技术,测定中膜VSMC,凋亡细胞及其相关蛋白,计算机图像分析并计算VSMC密度及凋亡指数。结果：与NA相比,AAA中膜VSMC密度降低,VSMC凋亡指数及其相关蛋白P53,P21明显增加,而bcl-2无显变化。结论：VSMC凋亡在细胞水平参与腹主动脉结构损伤与重构,促进AAA形成。     

血管平滑肌细胞凋亡在腹主动脉瘤形成机制中的作用

细胞凋亡（ａｐｏｐｔｏｓｉｓ）是机体在维持正常组织结构和生理功能过程中自然发生的细胞更替,它的异常可导致很多疾病,促进细胞凋亡可使正常细胞过多死亡,同样阻滞细胞凋亡的发生则可能导致恶性肿瘤。最近的研究表明细胞凋亡在正常和病变再塑形的血管壁中均起重要作用［１～２］。腹主动脉瘤（ａｂｄｏｍｉｎａｌａｏｒｔｉｃａｎｅｕｒｙｓｍ,ＡＡＡ）是血管壁再塑形的典型例子,以动脉壁逐渐变弱和渐进性扩张最后破裂为特征［３］。其主要病理变化包括：动脉壁中层弹力纤维的广泛断裂和退化、胶原纤维所占百分比相对过多、免疫及…     

终末期肾病血管钙化机制

终末期肾病(ESRD)患者血管钙化十分普遍,可导致心肌缺血、梗死等严重症状及死亡。深入研究其发生机制将有助于对其防治,减少ESRD患者心血管疾病(CVD)的发生,提高生存率。一、ESRD血管钙化的概述1998年美国ESRD患者的病死率为20%,其中透析患者的CVD病死率较普通人群高10～20倍犤1犦。CVD是ESRD患者的主要死因。Savage等犤2犦发现71%(17/24)患者存在明确的颈动脉和股动脉的钙化斑块。Goodman等犤3犦发现年轻透析患者(20～30岁)中,14(14/16)例存在冠状动脉钙化,其中9例在(20±3)个月后钙化积分平均增长近两倍;而正常人群中,仅3(3/…     

终末期肾病患者骨折影响因素的Meta分析

目的 系统分析终末期肾病患者发生骨折的相关因素。方法 计算机检索中国知网、维普、万方、中国生物医学数据库、PubMed以及EMBASE中自建库以来公开发表的相关文献。由2名研究员独立筛选文献、提取资料、纳入文献质量评价,采用Review Manager 5.3软件进行统计分析。结果 共纳入19篇文献,合并结果提取8个相关影响因素,其中6个危险因素：年龄（OR=1.28, 95%CI:1.20～1.37, P<0.01）、女性（OR=1.93, 95%CI:1.51～2.46, P<0.01）、碱性磷酸酶（OR=1.03,95%CI:1.01～1.05,P=0.008）、甲状旁腺激素（OR=1.07,95%CI:1.04～1.10,P<0.01）、糖尿病（OR=1.70,95%CI:1.44～2.02,P<0.01）、透析年限（OR=1.83,95%CI:1.20～2.80,P=0.005）;2个保护因素：血清白蛋白（OR=0.81,95%CI:0.69～0.96,P=0.02）和活性维生素D3(OR=0.33,95%CI:0.16～0.67,P=0.002)。结...     

胃肠道平滑肌肿瘤中血管形成与细胞凋亡的关系

通过检测胃肠道平滑肌肿瘤 (ｇａｓｔｒｏｉｎｔｅｓｔｉｎａｌｓｍｏｏｔｈｍｕｓｃｌｅｔｕｍｏｒｓ,ＧＩＳＭＴ)中微血管密度 (ＭＶＤ)、血管内皮细胞生长因子(ＶＥＧＦ)和细胞凋亡指数 (ＡＰＩ)、凋亡调控基因ｂｃｌ 2 ,以探讨血管生成、细胞凋亡及调控基因在ＧＩＳＭＴ发生发展过程中的作用及相互关系。资料与方法1.一般资料 :从中国医科大学第二临床学院 1977年～1996年间手术切除的ＧＩＳＭＴ病例中选择诊断明确、病历完整者 86例 ,其中男性 49例 ,女性 37例 ;年龄 1～ 77岁 ,平均48 6岁。肿瘤发生于食管 11例、胃 30例、小肠 2 7例…     

细胞外信号调节激酶对人血管平滑肌细胞的作用及其机制

目的观察磷酸化细胞外信号调节激酶(ERK)对离体培养的平滑肌细胞增殖、凋亡和表型改变的影响。方法培养人大隐静脉平滑肌细胞,实验组培养液中加入PD98059对磷酸化ERK进行阻断后,观测平滑肌细胞的增殖活性,凋亡细胞所占比例以及细胞表型,并与对照组比较。结果实验组与对照组相比,细胞增殖活性明显低于对照组(P<0.01),电镜和荧光染色测定凋亡细胞所占比例明显高于对照组(P<0.01),电镜检测合成型细胞所占比例明显低于对照组(P<0.01),收缩型细胞所占比例较对照组明显为高(P<0.01),α肌动蛋白免疫荧光化学检测显示处理组荧光表达强于对照组。结论对离体培养的人大隐静脉平滑肌细胞磷酸化ERK进行阻断后,平滑肌细胞的增殖受抑,凋亡增加,细胞由合成型向收缩型转变。     

终末期肾病血液透析患者Fetuin-A水平与血管钙化的关系

目的：观察终末期肾病（ESRD）血液透析患者血浆胎球蛋白A水平与血管钙化的关系,探讨血管钙化发生的相关因素。方法：收集40例ESRD患者的临床资料,检测血浆胎球蛋白A水平及对患者进行颈总动脉B超检查。所有患者的随访期限为12月。40例健康查体者为对照组。结果：ESRD患者血胎球蛋白A水平血液透析前较健康对照组明显降低（P〈0.001）,有血管钙化的患者胎球蛋白A水平明显低于无钙化者（P〈0.01）,经血液透析治疗1年后,血浆胎球蛋白A水平较透析前明显降低（P〈0.001）。相关性分析显示胎球蛋白A水平与C-反应蛋白（r=-0.35,P〈0.01）、血钙磷乘积（r=-0.37,P〈0.01）、血白蛋白（r=0.31,P〈0.05）,尤其血磷（r=-0.24,P〈0.01）相关。结论：ESRD患者经血液透析治疗后胎球蛋白A水平持续下降,血管钙化可能与血浆胎球蛋白A水平下降有关。     

23例终末期肾病患者透析时机影响因素的调查分析

目的 评估基层医院终末期肾病患者的透析时机,探讨过晚透析的影响因素及改进措施.方法 以郸城县人民医院透析科2007年7月1日至2009年7月1日开始透析的所有非糖尿病性终末期肾病患者为研究对象,分析其开始透析时血肌酐、肾小球滤过率、尿毒症症状、尿毒症心、脑血管并发症、首次透析情况,调查过晚透析的影响因素.结果 (1)进入透析时肾小球滤过率平均为(3.6±0.9)ml/min;(2)所有患者均有尿毒症并发症、营养状态下降,30.4%患者需急诊透析;(3)91.3%患者系透析时机过晚,其原因以经济原因多见,其次与就诊晚和认识不足有关.结论 基层医院开始血液透析的终末期肾病患者多数为透析时机过晚.经济原因、就诊过晚及认识不足是影响透析时机的关键因素.     

血管平滑肌细胞凋亡与尿毒症血管钙化

目的 探讨尿毒症患者血管平滑肌细胞（VSMC）凋亡与血管钙化之间的关系及其与临床指标的相关性。 方法 在43例尿毒症患者行动-静脉内瘘手术时留取桡动脉0.5~1 cm。用茜素红染色判断血管中膜钙化，并根据茜素红染色结果将患者分为钙化组和无钙化组。以肾功能正常、年龄相匹配的因外周血管疾病行血管手术的4例患者的动脉为对照组。采用DNA 片段原位缺口末端标记技术(TUNEL 法)检测上述3组血管平滑肌细胞凋亡情况，结果用凋亡指数表示。用免疫组化的方法检测凋亡相关基因p53、Bcl-2的表达情况。临床指标包括血钙、血磷、钙磷乘积、全段甲状旁腺素（iPTH）、血浆白蛋白（Alb）、血红蛋白（Hb）、总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白（LDL）、C反应蛋白（CRP）、丙二醛（MDA）、超氧化物歧化酶（SOD）。用Pearson相关分析计算凋亡指数与上述临床指标的相关性。 结果 尿毒症患者的血管中膜均可见到明显的VSMC凋亡，钙化组VSMC凋亡指数显著高于无钙化组（48.13±17.81比15.90±8.25，P ＜ 0.01）。而在对照组血管中膜只有极少量凋亡的VSMC。钙化组p53表达比无钙化组和对照组明显升高（31.22±14.25比20.67±7.35、4.17±2.61，P ＜ 0.01），Bcl-2表达比对照组明显降低（4.29±2.16比14.87±5.62，P ＜ 0.01)。VSMC凋亡指数与临床指标中的钙磷乘积、血磷和LDL水平呈正相关（P ＜ 0.05）。 结论 尿毒症患者桡动脉中膜存在VSMC凋亡，且VSMC凋亡发生在血管钙化之前。VSMC凋亡可能参与了尿毒症患者血管钙化的发生和发展。     

细胞凋亡与糖尿病肾病

糖尿病肾病 (ＤＮ)是糖尿病全身性微血管合并症之一 ,其早期改变为肾脏肥大、肾小球滤过率升高 ,肾小球及肾小管基底膜增厚 ,逐渐导致肾小球硬化 ,最终发生肾衰竭。其发病机理尚不十分清楚 ,目前研究证实细胞凋亡在其发生发展中起着重要作用。1　细胞凋亡的理化特征及相关基因细胞凋亡 (ａｐｏｐｔｏｓｉｓ)或细胞程序性死亡 (ｐｒｏ ｇｒａｍｍｅｄｃｅｌｌｄｅａｔｈ ,ＰＣＤ)是与细胞坏死截然不同的细胞死亡方式 ,是指在一定的生理或病理条件下 ,细胞遵循自身的规律 ,按步就班地走向死亡的过程。凋亡是维持细胞群体动态平衡的重要因素之…     

Smooth Muscle Cell Apoptosis in Primary Varicose Veins

OBJECTIVES: One of the important factors responsible for vessel wall remodelling is programmed cell death. In the paper the role of smooth muscle cell (SMC) apoptosis in primary varicose veins (PVV) is investigated. MATERIAL AND METHODS: Vein specimens were obtained from 40 patients with PVV. In each case proximal and distal (upper crural) great saphenous veins (GSV) were harvested. Morphometric computer assessed quantitative evaluation of SMCs, collagen and elastin content was carried out. Apoptotic cells were detected by TUNEL assay. The levels of p53, BAX, BCLl-2 and p21 mRNA expression were assessed by real time RT-QPCR and the presence of respective proteins in the vessel wall was confirmed by immunohistochemistry. RESULTS: In the proximal GSV segments a significant increase of p53, p21 and BCL-2 mRNA levels was found in PVV patients. In the distal segments BAX and BCL-2 expression levels were higher. Taking into account the patient age, elevated p53 mRNA expression level was noticed in the distal incompetent GSVs of young PVV patients. In this group a statistically significant increase in the apoptotic index (APIx) within the vein media was found which correlated positively with p53 mRNA expression level. There was no increase of the apoptotic activity in elderly patients that led to the structural changes increase. In proximal GSV segments, despite SMC amount reduction or presence of structural changes in perivalvular wall region, no increase of the APIx with was noticed. CONCLUSIONS: P53-related apoptosis is one of the regulatory mechanisms of vein wall homeostasis maintenance. During varicose vein development its activation is related to the early stages of the disease. In the further course, the down-regulation of the SMC apoptosis within the vein media leads to the structural changes increase. The reduction of the SMC population corresponding to an increase of p21 expression in proximal saphenous vein segments suggests that the cell cycle disturbances may lead to the 'weakness' of the proximal GSV wall. Valve injury is not the only factor leading to the varicose veins occurrence.     

Effects of L-arginine on Vascular Smooth Muscle Cell Proliferation and Apoptosis after Balloon Injury

Objective—Vital exhaustion (VE) and a hypercoagulable state both have been associated with coronary artery disease (CAD). Candidate mechanisms by which VE predicts CAD events are impaired fibrinolysis and inflammatory changes, the latter also affecting hemostasis. We investigated whether VE and inflammation would independently relate to hemostasis.

Design—Study participants were 217 (mean age?±?SD, 40?±?9 years) apparently healthy men and women working at an airplane manufacturing plant in Germany who completed the Shortened 9‐item VE Maastricht Questionnaire. All subjects had a set of classic cardiovascular risk factors assessed, and plasma levels of fibrin D‐dimer, type I plasminogen activator inhibitor (PAI‐1) antigen, C‐reactive protein (CRP), and tumor necrosis factor (TNF)‐α were measured.

Results—PAI‐1 correlated with VE (r?=?0.18, p?=?0.009), CRP (r?=?0.20, p?=?0.004), and TNF‐α (r?=?0.18, p?=?0.009); D‐dimer correlated with CRP (r?=?0.16, p?=?0.018). In linear regression analyses, VE and TNF‐α independently explained 2 and 1%, respectively, of the variance in PAI‐1.

Conclusion—Our study corroborates previous findings on impaired fibrinolysis in VE. The findings suggest that VE and inflammation may impair fibrinolysis by different pathways, and independently of traditional cardiovascular risk factors.     

内放射对兔颈动脉术后中膜平滑肌细胞增殖与凋亡的影响

目的　通过血管内放射防治颈动脉内膜剥脱术 (CEA)后再狭窄 ,动态观察其对中膜平滑肌细胞(SMC)凋亡和增殖的影响 ,并初步探讨其防治再狭窄的可能机理。方法　 4 0只兔行CEA后 ,随机分为 4组 ,分别给予 0Gy、10Gy、2 0Gy及 4 0Gy放射剂量的3 2 P ,于术后 3d、7d、14d、2 8d及 5 6d分别处死动物 ,取出血管标本进行病理组织学分析。结果　术后除 3d及 7d外 ,对照组发生明显内、中膜增生及管腔狭窄 ,而内放射组可见内、中膜增生明显受抑制 ,管腔面积缩小显著减轻 (P<0 .0 5 )。SMC凋亡率和细胞增殖核抗原 (PCNA)阳性细胞均于术后 3d增加 ,7d时达高峰 ,治疗组与对照组比较差异有显著性意义 (P<0 .0 1) ;2 0Gy和 4 0Gy的作用明显强于 10Gy组 (P<0 .0 1)。结论　内放射治疗可能是通过抑制SMC增殖、迁移 ,诱导SMC凋亡 ,防治再狭窄 ,并且随剂量呈递增效应 ;球囊损伤后早期以细胞增生和凋亡为主要表现 ,晚期表现为内、中膜面积的增加     

Vascular Smooth Muscle Cell Apoptosis Promotes Transplant Arteriosclerosis Through Inducing the Production of SDF‐1α

Transplant arteriosclerosis is a leading cause of late allograft loss. Medial smooth muscle cell (SMC) apoptosis is considered to be an important event in transplant arteriosclerosis. However, the precise contribution of medial SMC apoptosis to transplant arteriosclerosis and the underlying mechanisms remain unclear. We transferred wild‐type p53 to induce apoptosis of cultured SMCs. We found that apoptosis induces the production of SDF‐1α from apoptotic and neighboring viable cells, resulting in increased SDF‐1α in the culture media. Conditioned media from Ltv‐p53‐transferred SMCs activated PI3K/Akt/mTOR and MAPK/Erk signaling in a SDF‐1α‐dependent manner and thereby promoted mesenchymal stem cell (MSC) migration and proliferation. In a rat aorta transplantation model, lentivirus‐mediated BclxL transfer selectively inhibits medial SMC apoptosis in aortic allografts, resulting in a remarkable decrease of SDF‐1α both in allograft media and in blood plasma, associated with diminished recruitment of CD90⁺CD105⁺ double‐positive cells and impaired neointimal formation. Systemic administration of rapamycin or PD98059 also attenuated MSC recruitment and neointimal formation in the aortic allografts. These results suggest that medial SMC apoptosis is critical for the development of transplant arteriosclerosis through inducing SDF‐1α production and that MSC recruitment represents a major component of vascular remodeling, constituting a relevant target and mechanism for therapeutic interventions.     

坏死细胞释放IL-1对血管平滑肌细胞增殖及炎性反应的影响

目的 研究坏死血管平滑肌细胞对周围正常细胞增殖及炎性因子分泌的影响,并探讨其可能的作用机制.方法 无血清低糖低氧条件下培养细胞,建立细胞坏死模型,收集坏死细胞培养上清液,用于干预正常血管平滑肌细胞.实验设坏死上清组、IL-1组、坏死上清+IL-1RA组和对照组.MTT法检测各组细胞增殖情况;RT-PCR检测各组MCP-1、IL-6和IL-8 mRNA的表达;ELISA检测各组MCP-1、IL-6和IL-8的分泌;Western blot检测各组NF-kB的激活情况.结果 NCS组和IL-1组细胞增殖能力均显著高于对照组(P＜0.01),NCS+IL-1 β组细胞增殖能力显著低于NCS组和IL-1组(P＜0.01);NCS组和IL-1 β组MCP-1、IL-6、IL-8 mRNA的表达均显著高于对照组(P＜0.05),NCS+IL-1RA组MCP-1、IL-6、IL-8 mRNA的表达均显著低于NCS组和IL-1组(P＜0.05);NCS组和IL-1 β组的MCP-1、IL-6及IL-8蛋白的分泌均显著高于对照组(P＜0.05),NCS+IL-1RA组MCP-1、IL-6和IL-8蛋白的分泌显著低于NCS组及IL-1组(P＜0.05);NCS组和IL-1组NF-kB的活性显著高于对照组(P＜0.05),与NCS组和IL-1组相比NCS+IL-1RA组NF-kB的活性显著降低(P＜0.05).结论 坏死血管平滑肌细胞能够促进正常细胞的增殖,诱导NF-kB的激活以及炎性因子的释放,并且这种促进作用可能与IL-1的大量释放有关.     

Comparison of Vascular Smooth Muscle Cell Apoptosis and Fibrous Cap Morphology in Symptomatic and Asymptomatic Carotid Artery Disease

The biological cascades that lead to carotid plaque disruptions and symptoms are largely unknown. Certain cellular events within the plaque might be responsible for destabilizing the plaque, though the popular belief is that the plaque size is directly related to symptoms. The aim of our study was to assess the morphology of the fibrous cap and apoptosis in the plaque and compare these two pathological features in symptomatic and asymptomatic carotid artery disease. Our work was carried out in plaques obtained following carotid endarterectomy performed for symptomatic disease (including hemispheric transient ischemic attacks, amaurosis fugax, or stroke) or asymptomatic high-grade severe stenosis. Scion images of Gomori's stained sections were used to measure fibrous cap thickness and area. TUNEL assay was performed to assess the extent of apoptosis. The results indicated that the area of the fibrous cap did not significantly correlate with the presence of symptoms. There was a higher percentage of apoptotic nuclei and the thinner fibrous cap in symptomatic plaques than in asymptomatic plaques. This finding suggests that these factors might be involved in destabilizing plaque, causing rupture and leading to symptomatic carotid disease.     

白细胞介素-10腺病毒载体制备及其在血管平滑肌细胞的表达

目的构建人白细胞介素(human interleukin,hIL)-10和绿色荧光蛋白(green fluorescent protein,GFP)双表达腺病毒载体,观察目的基因在血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的表达。方法以T载体(pUCm-T/hIL-10cDNA)为模板,PCR扩增hIL-10cDNA,连入穿梭质粒,卡那霉素抗性筛选,NotⅠ、XholⅠ酶切鉴定,测序,局部序列搜索工具(BLAST)分析序列正确,PmeⅠ酶切,电穿孔单独转化BJ5183-AD-1,转化XL10-Gold扩增,PacⅠ酶切,AD-293细胞包装,PCR鉴定,测定病毒滴度,脂质体介导转染VSMCs,荧光显微镜下观察GFP的表达,酶联免疫吸附试验(ELISA)检测培养上清液中hIL-10的含量。结果重组穿梭质粒构建正确,同源重组后PacⅠ酶切获得30kb和3kb片段,重组病毒滴度为3×1010efu/ml,PCR检测含目的基因,转染的VSMCs可见绿色荧光,hIL-10含量为25ng/106个细胞。结论构建的双表达载体于VSMCs成功表达,为血管内膜增生的基因治疗打下基础。     

低氧对SD大鼠阴茎海绵体平滑肌细胞凋亡的影响

目的:探讨低氧和SD大鼠阴茎海绵体平滑肌细胞(CCSMC)凋亡的关系。方法:体外培养CCSMC,免疫组化鉴定细胞;低氧(1%O2浓度)干预12、24、48、72 h,常规氧浓度作为对照,流式细胞术测定细胞周期变化和凋亡情况。结果:体外培养的CCSMC生长良好,抗平滑肌α-肌动蛋白单克隆抗体免疫组化DAB法染色阳性。流式细胞术检测CCSMC G0/G1期在48 h内细胞比例逐渐增加,后下降;S期细胞比例与G0/G1期呈相反趋势;G2/M期细胞比例无明显规律。结论:CCSMC在低氧环境下,随时间的延长凋亡程度加重,48 h达到最大值,进一步延长时间细胞裂解,不能加重凋亡。     

OPG对高磷诱导的牛血管平滑肌细胞钙化的作用

目的：研究护骨素（OPG）对高磷诱导的牛血管平滑肌细胞钙化是否具有抑制作用。方法：用含不同磷（Pi）浓度（1.4～2.0mmol/L）的培养基体外培养牛血管平滑肌细胞,观察细胞钙沉积及OPG的表达;在高磷（2.0mmol/L）培养基中加入不同浓度外源性OPG测定细胞钙化值的变化。用甲ó-酚肽络合酮法测定钙化量,BCA法（二喹啉甲酸检测法）测蛋白含量,并以细胞蛋白含量标化钙化量。Western Blotting法测定OPG蛋白表达。结果：（1）培养基磷浓度1.4mmol/L组细胞仅有少量钙沉积,随着培养基磷浓度增高及培养时间的延长,细胞钙沉积增加,第9天时,iP2.0mmol/L组与iP1.4mmol/L组钙沉积[（138.00±12.53）μg/mg protein比（22.67±2.52）μg/mg protein]差异有统计学意义。（2）随着培养基磷浓度增高OPG蛋白表达量相应增加。（3）随着培养基中OPG浓度的增高细胞钙化有减少趋势,OPG浓度50ng/ml和100ng/ml组钙化分别为（82.67±5.79）μg/mg protein、（60.67±7.36）μg/mg protein,与单纯高磷组比较其差异有统计学意义（P〈0.05）。结论：高磷培养基能诱导血管平滑肌发生钙化和OPG表达,且与磷浓度和作用时间有关,外源性OPG能抑制高磷诱导的细胞钙化的产生,而且这种抑制作用具剂量依赖性。     

RNA干扰对兔血管平滑肌细胞bcl-2基因表达的影响

目的研究RNA干扰(RNAi)对兔血管平滑肌细胞(VSMCs)bcl-2基因表达的干扰效应。方法用脂质体法将构建的装载靶向bcl-2基因小干扰性RNA(siRNA)的表达载体(pshRNA-bcl-2质粒)转染VSMCs(基因组),以转染空载体的细胞和加DMEM的空白细胞作为对照,采用半定量RT-PCR和Westernblot法检测bcl-2基因的表达,用MTT法检测VSMCs生长情况。结果转染siRNA的表达载体可以抑制内源性bcl-2基因在转录和转译水平上的表达,基因组bcl-2的mRNA和蛋白表达较空载体组和空白对照组明显减少(P<0.01);基因组的VSMCs生长也较空载体组和空白对照组明显受到抑制(P<0.01)。结论载体介导的RNAi技术可明显抑制内源性bcl-2基因的表达和VSMCs生长。