冠状动脉斑块中炎性细胞和平滑肌细胞的定量及其与斑块稳定性的关系

目的　探讨稳定及不稳定斑块中巨噬细胞、Ｔ淋巴细胞及平滑肌细胞数量的差异及其与斑块稳定性的关系。方法　将急性心肌梗死死亡病例的尸检冠状动脉完整剥离后固定、脱钙、连续取材并常规切片及ＨＥ染色,按有无脂质中心及其大小将斑块分为稳定斑块（ 斑块无或仅有较小的脂质中心） 及不稳定斑块（ 脂质中心超过斑块的４０ ％ ）。选取２ 种斑块各１６３ 个,用ＣＤ６８ 、肌动蛋白、ＵＣＨＬ－ １ 抗体分别对巨噬细胞、平滑肌细胞、Ｔ淋巴细胞进行免疫组化染色,并通过计算机图像分析系统对２ 种斑块纤维帽中的３ 种细胞进行定量。结果　不稳定斑块纤维帽中巨噬细胞及Ｔ淋巴细胞的面积百分比分别为８９．６％ 和３２．５％ ,稳定斑块分别为１８．３％ 和７．２ ％ ,差异均具有显著性（ Ｐ＜０．０１） 。而平滑肌细胞在稳定斑块纤维帽中的面积百分比明显大于不稳定斑块,分别为６８．４ ％ 和２１ ．８％ ,差异具有显著性（Ｐ＜ ０．０５） 。结论　稳定斑块与不稳定斑块不仅脂质中心大小不同,其纤维帽中炎细胞和平滑肌细胞的数量也不同,粥样坏死中心越大,其表面纤维帽中的巨噬细胞、Ｔ淋巴细胞就越多,而平滑肌细胞则越少。脂质中心大小和纤维帽中炎细胞数量是决定斑块稳定性的主要因素。     

兔不稳定斑块的18F-FDG PET/CT显像及其病理学检测

 目的 探讨氟18标记的脱氧葡萄糖 (18F-FDG) PET/CT无创检测不稳定斑块的可行性。方法 用新西兰雄性大白兔建立动脉粥样硬化模型,注射18F-FDG后行腹主动脉PET/CT成像,离体动脉数码照相,测定动脉片段的放射强度,选取不稳定和稳定斑块各30块,比较其放射强度,采用免疫组化方法观察斑块中的巨噬细胞和平滑肌细胞并计数。结果 PET/CT活体成像可见实验组沿腹主动脉走行的不均匀的放射性分布,与离体动脉的斑块分布基本一致;不稳定斑块组的靶-非靶比值和巨噬细胞数明显高于稳定斑块组(P＜0.01),而平滑肌细胞数明显降低(P＜0.01);靶-非靶比值与斑块中的巨噬细胞数呈正相关(r=0.815,P＜0.01),而与平滑肌细胞数呈负相关(r=-0.684,P＜0.01)。结论18F-FDG PET/CT无创检测实验性不稳定斑块具有一定的可行性。     

缺氧时肺动脉内皮细胞与肺动脉平滑肌细胞相互作用对细胞内 …

目的：观察了培养的小牛肺动脉内皮细胞（ＰＡＥＣ）和肺动脉平滑肌细胞（ＰＡＳＭ）于缺氧时对细胞内环核苷酸的影响。结果：ＰＡＥＣ和ＵＰＡＳＭ共培养２４ｈ，ＰＡＥＣ细胞内ｃＡＭＰ含量显著降低（Ｐ〈０．０１），而ＰＡＳＭ细胞内ｃＡＭＰ含量显著增加（Ｐ〈０．０１），二种细胞内ｃＧＭＰ含量显著增加（Ｐ〈０．０１）。缺氧对二种细胞内ｃＧＭＰ含量无显著影响，但能增加ＰＡＳＭ的ｃＧＭＰ含量（Ｐ〈０．０１），降低ＰＡ     

培养的大鼠血管平滑肌细胞中bFGF基因表达与血管紧张素Ⅱ …

采用Ｎｏｒｔｈｅｒｎ杂交检测培养的自发性高血压大鼠（ＳＨＲ）和正常血压ＷＫＹ大鼠的血管平滑肌细胞（ＶＳＭＣ）中碱性成纤维细胞生长因子（ｂＦＧＦ）基因表达，用紫外法和放免法分别测定培养液中血管紧张素转换酶（ＡＣＥ）活性和血管紧张素Ⅱ（ＡｎｇⅡ）含量，发现ＡｎｇⅡ能明显促进ＶＳＭＣ中ｂＦＧＦ基因表达，而ｂＦＧＦ则能明显诱导ＡＣＥ活性和提高ＡｎｇⅡ释放，且ＳＨＲＶＳＭＣ的ｂＦＧＦ基因表达，ＡＣＥ活性和Ａ     

球囊损伤血管内皮后平滑肌细胞凋亡及相关基因表达 …

目的：探讨球囊损伤血管内皮后平滑肌细胞（ＳＭＣ）凋亡的机制。方法：采用末端脱氧核苷酸转移酶介导的三磷酸脱氧尿嘧啶缺口末端标记法（ＴＵＮＥＬ）和免疫组织化学技术检测球囊损伤内皮后血管平滑肌细胞（ＶＳＭＣ）凋亡及Ｂａｘ,Ｂｃｌ－２蛋白的变化,结果：球囊损伤内皮后第３ｄ,血管中层出现凋亡的ＳＭＣ;损伤后第７ｄ,内膜和中层ＳＭＣ凋亡率最高,凋亡的ＳＭＣ主要分布内膜层;以后爱渐降低,至损伤后第２８ｄ,仅内膜     

冠状动脉粥样硬化斑块内血管新生与斑块稳定的关系

目的　比较稳定斑块和不稳定斑块内新生血管形态学分布和数量上的差异 ,探讨新生血管与斑块稳定的关系。方法　从死亡前有急性冠脉综合征发生的 5 2例尸检冠状动脉标本中选取晚期动脉粥样硬化病变的组织块共 92 2块 ,以脂质核心面积是否大于斑块面积的 4 0 %为标准将其分为不稳定斑块组 (15 3块 )和稳定斑块组 (76 9块 ) ,比较两组斑块内新生血管的检出率。由两组随机选取各 4 0个组织块进行第八因子相关抗原抗体的免疫组织化学染色 ,观察阳性物质在斑块内的分布特点并通过计算机图像分析系统进行量化分析。结果　不稳定斑块组新生血管检出率 (80 4 % )显著高于稳定斑块组 (6 6 6 % ,P <0 0 1)。新生血管主要分布在斑块的肩部和基底部 ,纤维帽相对较少 ;不稳定斑块组各部位的新生血管最大密度 [肩部 :(2 2 16± 19 96 )个 /mm2 ,基底部 :(2 1 6 8± 2 0 4 4 )个 /mm2 ,纤维帽 :(3 80± 5 32 )个 /mm2 ]均显著大于稳定斑块组的相应部位 [肩部 :(10 0 4± 11 5 2 )个 /mm2 ;基底部 :(9 6 8± 11 5 2 )个 /mm2 ;纤维帽 :(1 4 8± 2 2 8)个 /mm2 ,P <0 0 5 ]。结论　不稳定斑块组较稳定斑块组新生血管检出率和密度均显著增高 ,提示新生血管与斑块的不稳定性密切相关。除目前较为公认的脂质核心大小     

动脉粥样硬化斑块形成时血管平滑肌细胞增生相关基因的cDNA全 …

目的 克隆动脉粥样硬化斑块形成血管平滑肌细胞（ＳＭＣ）增生相关基因的ｃＤＮＡ全长,并对其功能进行初步探讨。方法 选用氧化低密度脂蛋白作为刺激物作用于培养的人主动脉ＳＭＣ,利用减除杂交技术构建减除文库,克隆相关基因片段,在全长文库构建的基础上筛选相关基因的ｃＤＮＡ全长。并进行原核系统内蛋白表达及量效关系检测。结果 发现了４个新基因片段,克隆了一个新基因ｃＤＮＡ全长,该基因在原核系统内有约相对分子质量     

骨桥蛋白和骨连结蛋白在冠状动脉粥样硬化斑块中的表达及与钙化的关系

目的 :探讨骨桥蛋白 (Osteopontin ,OPN)和骨连结蛋白 (Osteonectin ,ON)在冠状动脉粥样硬化斑块中的表达情况及与钙化的关系。方法 :从 1 3例压力灌注固定后的心脏标本中分离 2 6支冠状动脉 ,取其中的 2 60个血管段进行组织切片 ,HE染色。用LF1 2 3、LF3 7、CD68、α actin抗体分别对OPN、ON、巨噬细胞、平滑肌细胞进行免疫组化染色 ,Leica图像分析仪观察斑块的OPN、ON以及钙化和两种细胞的染色程度。结果 :钙化斑块内均有OPN表达 ,OPN的表达程度随钙化加重而增加 (x2 =3 5 0 .0 79,P <0 .0 0 1 ) ;ON在正常和粥样硬化的冠状动脉基本不表达或少数弱阳性 ;OPN的表达程度与CD68和α actin的表达水平之间有显著关联性 ,即随着OPN表达程度的增加 ,CD68的表达显著增强 ,巨噬细胞数增多 (x2 =1 82 .83 9,P <0 .0 0 1 ) ,α actin的表达显著减弱 ,平滑肌细胞数减少 (x2 =5 8.875 ,P <0 .0 0 1 )。结论 :OPN参与冠状动脉钙化的调节 ,其主要来源于巨噬细胞和巨噬细胞源性的泡沫细胞。ON在冠状动脉钙化中的作用不明     

缺氧对肺动脉内皮细胞和平滑肌细胞5—羟色胺转载体基因表达 …

目的：探讨无氧（０％Ｏ２＋９５％Ｎ２＋５％ＣＯ２）和低氧（２．５￣３％＋９％Ｎ２＋５％ＣＯ２）对新生小牛肺动脉内皮细胞（ＰＡＥＣ）和肺动脉平滑肌细胞（ＰＡＳＭ）５－羟色胺转载体基因表达的影响。方法：应用细胞培养，核到分子杂交技术。结果：无氧组和低氧组（１．５ｈ、３ｈ、６ｈ）ＰＡＥＣ５－羟色胺转载体ｍＲＮＡ表达显著同于常氧组（Ｐ〈０．０５），而无氧和低氧１２ｈ至４８ｈ对ＰＡＥＣ５－羟色胺转载体ｍＲＮ     

Identification of macrophages and smooth muscle cells with monoclonal antibodies in the human atherosclerotic plaque

Summary Sections of human atherosclerotic plaques, obtained from 21 autopsy cases with various degrees of atherosclerosis, were stained with the indirect immunoperoxidase technique using specific monoclonal antibodies against macrophages and smooth muscle cells. Distinctive results were found in differing stages: Single blood monocytes were observed in diffuse intimal thickening and the foam cells seen in fatty streaks were mostly identified as mature tissue macrophages, while only very few blood monocytes were present. The spindle cells observed in fibroelastic plaques showed positive reactions to antibodies against desmin, which points to their derivation from smooth muscle cells, whereas only a few macrophage-derived foam cells were seen in these lesions. In the complicated lesions the majority of foam cells were macrophage-derived, but there was also a small number of foam cells positive to antibodies against desmin, suggesting a smooth muscle cell derivation. - Our results confirm that in human atherosclerotic plaques the majority of the foam cells are obviously macrophage-derived, which emphasizes the important role of macrophages in the morphogenesis of these lesions.Supported by Landesamt für Forschung Nordrhein-Westfalen     

Heterogeneity of smooth muscle cells in atheromatous plaque of human aorta.

This study was undertaken to investigate the expression of cytoskeletal proteins and the ultrastructure of cells in normal intima and atheromatous plaque of human aorta. It has been established, using double-labeling immunofluorescence, that smooth muscle cells (SMC) in normal aortic intima contain myosin, vimentin, and alpha-actin but do not react with antibodies against desmin. In contrast, 7 of 28 atherosclerotic plaques contained many cells expressing desmin in addition to the other cytoskeletal proteins characteristic of normal intima SMC. These cells were localized predominantly in the plaque cap and had the ultrastructural features of modulated SMC, ie, well-developed endoplasmic reticulum and Golgi apparatus. Besides, some cells in the 13 atherosclerotic plaques proved to be myosin, alpha actin, and desmin negative but contained vimentin and actin as revealed by fluorescent phalloidin. These cells were found in the immediate proximity of atheromatous material and reacted with a monoclonal antibody specific to SMC surface protein (11G10) but not with monoclonal anti-muscle actin (HHF35) and anti-macrophage (HAM56) antibodies. Electron microscopy of this plaque zone revealed that the cytoplasm of these cells was filled with rough endoplasmic reticulum and a developed Golgi complex. At the same time, a certain proportion of cells in this region retained morphologic features of differentiated SMC such as the presence of a basal lamina and myofilament bundles. The revealed peculiarities of cytoskeletal protein expression and the ultrastructure of cells in human aortic atherosclerotic plaques may be explained by a phenotypic modulation of vascular SMC.     

Cytokeratins in smooth muscle cells and smooth muscle tumours

A keratin positive metastatic leiomyosarcoma in the lung, which resulted in diagnostic error, is reported. The results of additional studies of 17 benign and malignant leiomyogenic tumours with various keratin antibodies are presented and discussed in the light of recent bibliographical data.     

Electrical coupling between smooth muscle cells and endothelial cells in pig coronary arteries

 The control of smooth muscle cells by endothelial cells has been well established by the identification of vasoactive factors released by the endothelial cells. In contrast, the possibility that smooth muscle cells influence the endothelial cells has been considered rarely. Some results suggest possible electrical communication between the smooth muscle and the endothelial cells but proof is lacking. We therefore tested for electrotonic conduction of signals from smooth muscle cells to endothelial cells. The endothelium was removed from half of a strip of porcine coronary artery. In a partitioned chamber, rectangular hyperpolarization or depolarization was applied to the de-endothelialized region by field stimulation. The resulting membrane potential changes in the smooth muscle cells spread electrotonically along the media into the area with intact endothelium. We recorded from endothelial cells to determine whether this electrical signal spreads into endothelial cells. Hyperpolarization or depolarization initiated in smooth muscle cells was recorded consistently in endothelial cells. This demonstrates a functional electrotonic propagation from smooth muscle to endothelial cells. Received: 23 May 1996 / Received after revision: 3 September 1996 / Accepted 16 September 1996     

缝隙连接在血管内皮和平滑肌细胞中的作用及其与高血压的关系

细胞间的直接通讯--缝隙连接在控制和协调血管功能方面起重要作用。缝隙连接是沟通相邻细胞质膜的唯一一类通道,为离子、小分子代谢物质、第二信使等细胞间信号分子的直接交换提供一个通道。血管内皮细胞和平滑肌细胞表达多种类型缝隙连接蛋白(connexin,Cx),多种Cx 在心血管系统中的共同表达并协调一致地工作,对心血管系统的发育、平滑肌和内皮细胞功能的整合、以及对血管壁细胞功能协调等起着重要作用。在血管发生疾病时Cx表达相应发生改变,以与缝隙连接调节血管紧张度和动脉血压的作用相一致。本综述讨论了在血管内皮和平滑肌细胞在生理和病理情况下(以高血压为例)缝隙连接对血管结构和功能的影响。     

The response to percutaneous transluminal coronary angioplasty: An ultrastructural study of smooth muscle cells and endothelial cells

We investigated the ultrastructure of the repair tissue formed after percutaneous transluminal coronary angioplasty (PTCA). Four hearts (6 coronary arteries) were investigated; 5 arteries after single PTCA and 1 after repeated PTCA. In the earliest lesion (5 days after PTCA), all smooth muscle cells were of a synthetic phenotype, with abundant rough endoplasmic reticulum and few myofilaments, whereas the oldest lesion (259 days after PTCA) was composed of contractile smooth muscle cells, characterized by abundant myofilaments and small amounts of rough endoplasmic reticulum. In lesions taken at intermediate times (74 and 77 days), the smooth muscle cells were of an intermediate phenotype, with rough endoplasmic reticulum and myofilaments present in approximately equal portions. Regenerating endothelial cells observed at 74 days after PTCA were characterized by a rounded shape and a paucity of intercellular connections in the form of simple junctions, which suggested an immature state. At 259 days after PTCA, the endothelial cells were more mature, as indicated by a flat shape and the presence of many tight junctions and a complete basement membrane. The findings suggest a relation between the stage of endothelial cell regeneration and the phenotype of the smooth muscle cells.     

Interactions of coronary artery smooth muscle cells with 3D porous polyurethane scaffolds

One strategy in vascular tissue engineering is the design of hybrid vascular substitutes where vascular cells infiltrate biostable porous scaffolds that provides favorable environment for guided cell repopulation and acts as a mechanically supporting layer after the tissue regeneration process. The aim of the present work was to study the interaction of human coronary artery smooth muscle cells (HCASMC) with 3D porous polyurethane scaffolds. We therefore fabricated porous and highly interconnected 3D polyurethane scaffolds that can promote HCASMC attachment, proliferation, and migration. SEM and microCT studies of the fabricated scaffolds showed that the current scaffolds had highly open and interconnected pore structures, with an average porosity of 84%. HCASMC interaction on polyurethane films revealed that cells adhere and express specific marker proteins (vinculin and h-caldesmon). This expression was further enhanced by coating the polyurethane with Matrigel. On uncoated 3D scaffolds, dense spherical aggregates of cells were often encountered with little adhesion of individual cells alongside the struts of the scaffold, independent of the porogens used. In contrast, when cultured on Matrigel-coated scaffolds, cell numbers quickly increased after 14 days and spread along the entire scaffold. At the upper scaffold surface, elongated cells were seen adhering to one another and also to the scaffold surface. These cells were elongated, aligned in parallel and contained abundant F-actin bundles suggesting a differentiated contractile phenotype. Deep into the scaffold, cells were encountered that formed actin-rich lamellipodial extensions spreading along the strut and lacked stress fibers, suggesting active cell migration along the substrate.