Monocyte-macrophage infiltration in portions of atherosclerotic lesions of the human aorta]

Human thoracic aorta was taken from trauma victims aged from 6 to 40 years. Cell composition of the intimal layer was investigated in the lesion predisposed (LP) areas of the aorta and in the lesion resistant (LR) areas. By double immunofluorescent staining with mono and polyclonal antibodies to monocytes/macrophages (Mn/Mph) and smooth muscle cells (SMC) the presence of Mn/Mph was revealed in all aortas studied. The number of these cells was 2-6 fold higher in LP areas, compared to LR areas in all persons over 21 years of age. Scanning electron microscopy revealed Mn on the luminal surface of the vessel the number of which was frequently higher in LP areas. Mn/Mph in LP areas were heterogeneous in their structure depending on the depth of their localization in the intima. The revealed Mn/Mph infiltration in aortas of young subjects may be the earliest manifestation of the atherosclerotic lesion.     

人主动脉粥样硬化病变中修饰脂蛋白的研究

目的研究人粥样硬化病变主动脉中丙二醛（ＭＤＡ）和４羟基壬烯醛（ＨＮＥ）修饰载脂蛋白Ｂ（ａｐｏＢ）的分布特点、含量和理化特性,并与脂蛋白（ａ）［Ｌｐ（ａ）］和ａｐｏＢ的分布进行比较。方法应用免疫组织化学、电镜、免疫电镜以及生物化学等方法进行定性或定量分析。结果ＭＤＡ和ＨＮＥ－ａｐｏＢ在细胞外基质中的分布与Ｌｐ（ａ）和ａｐｏＢ一致,但在泡沫细胞内则有不同。ＭＤＡ－ａｐｏＢ和ＨＮＥ－ａｐｏＢ在泡沫细胞内呈环状、孔状分布,与蜡样色素相似。病变动脉内膜低密度脂蛋白的超微结构、化学成分及电泳行为均与体外修饰脂蛋白相似,其醛类修饰ａｐｏＢ含量明显高于无病变者。结论脂蛋白的氧化性修饰是动脉粥样硬化发生的必要条件。     

Interstitial collagens in fibrous atherosclerotic lesions of human aorta

The present study was undertaken to clarify the existing controversy on the collagenous content and composition of human fibrous atherosclerotic versus normal aortic tissues. Several analytic procedures (slab gel electrophoresis; cyanogen bromide peptide mapping; high performance liquid chromatography; ion exchange chromatography) revealed that the amount of the interstitial collagens, i.e. types I and III, was similar in fibrous atherosclerotic lesions and control tissues (70% and 30% respectively). Moreover, when fibrous lesions were analyzed as serial fractions there was a uniform distribution of type I and type III throughout the lesion. Small increases in type III were observed only beneath the lesion where it interfaced with the normal media. The results suggest that contrary to some previous studies no major shifts in the ratio of the interstitial collagens are evident in atherosclerotic lesions as compared to normal intima-media preparations.     

Ciliated smooth muscle cells in atherosclerotic lesions of human aorta

One or two rudimentary cilia were observed by electron microscopy in smooth muscle cells (SMCs) of fatty dots and streaks, but not in normal intima of human aorta. Similar organelles are known to occur in many cell types and various species, but to the best of the author's knowledge, have never been found in the SMCs of arterial or other tissues of man in health and disease; recently, they were reported to be present in the SMCs of experimental atherosclerosis in rabbits. The rudimentary cilia observed in this study had a "9 + 0" axoneme (microtubular complex) and differed also in other aspects from the classical cilia. Semiserial sections of SMCs containing a diplosome disclosed that on several occasions both of its constituent centrioles gave rise to rudimentary cilia. SMCs containing cilia or their basal bodies were observed more often in the human than in experimental atherosclerotic lesions. Whereas the function and significance of the rudimentary cilia remain largely unknown, the current theory proposes that a sudden transformation from a mitotic replicative to a nonmitotic structured tissue "diverts" centrioles to the formation of these unusual organelles. It is conceivable that rudimentary cilia could serve as morphological indicators of aborted mitosis in human atherosclerotic lesions.     

Microcalcifications in early intimal lesions of atherosclerotic human coronary arteries

Although calcium (Ca) precipitation may play a pathogenic role in atherosclerosis, information on temporal patterns of microcalcifications in human coronary arteries, their relation to expression of calcification-regulating proteins, and colocalization with iron (Fe) and zinc (Zn) is scarce. Human coronary arteries were analyzed post mortem with a proton microprobe for element concentrations and stained (immuno)histochemically for morphological and calcification-regulating proteins. Microcalcifications were occasionally observed in preatheroma type I atherosclerotic intimal lesions. Their abundance increased in type II, III, and IV lesions. Moreover, their appearance preceded increased expression of calcification-regulating proteins, such as osteocalcin and bone morphogenetic protein-2. In contrast, their presence coincided with increased expression of uncarboxylated matrix Gla protein (MGP), whereas the content of carboxylated MGP was increased in type III and IV lesions, indicating delayed posttranslational conversion of biologically inactive into active MGP. Ca/phosphorus ratios of the microcalcifications varied from 1.6 to 3.0, including amorphous Ca phosphates. Approximately 75% of microcalcifications colocalized with the accumulation of Fe and Zn. We conclude that Ca microprecipitation occurs in the early stages of atherosclerosis, inferring a pathogenic role in the sequel of events, resulting in overt atherosclerotic lesions. Microcalcifications may be caused by local events triggering the precipitation of Ca rather than by increased expression of calcification-regulating proteins. The high degree of colocalization with Fe and Zn suggests a mutual relationship between these trace elements and early deposition of Ca salts.     

Intercellular contacts in the media of the thoracic aorta of rat fetuses treated with beta-aminopropionitrile

Beta-aminopropionitrile (BAPN) alters the scleroproteins of the arterial media in a way which permits an increase in the stress (force/unit area) applied to smooth muscle cells at a given pressure. Pregnant Sprague-Dawley rats were treated with BAPN and the thoracic aortas of exposed fetuses were studied by electron microscopy and image analysis. Transmission electron microscopy of the media of the thoracic aorta revealed that the types of intercellular contacts seen in the controls and in the BAPN group were intermediate junctions (IJs), nexus junctions (NJs), simple appositions (SAs), and interdigitations (Ids) as in adult vessels. The BAPN-treated animals showed an increased density of IJs, SAs, and Ids when compared with controls but the NJs demonstrated no change. Load-sensitive intercellular contacts have been considered to be the major route by which stress is applied across cells, and the increased number in the BAPN group probably represents a response of the vessel wall to increased tangential tension.     

Analysis of the intermediate and basement membrane collagens in fibrous atherosclerotic lesions of human aorta

The collagens of fibrous atherosclerotic lesions of human aortae obtained at post mortem examination were compared with those of normal intima-media preparations. Assessed quantitatively, pepsin-solubilized types IV, V and VI collagens decreased in relation to types I and III in preparations from lesions as compared to values for controls. The type V collagen in both tissues were composed of alpha 1 (V) and alpha 2 (V) chains in a 2:1 ratio. A novel ("V") collagen polypeptide identical in size to the alpha 1 (V) chain was identified in association with the interstitial collagen fraction in both tissue types. This chain had unique solubility characteristics and cyanogen bromide peptide composition. The exact relation of this polypeptide to the other collagens is not known, but it is possible that it accounts for the reported fluctuations in type V chains in aortic tissues.     

脂蛋白（a）在主动脉粥样硬化病变中的定位与定量研究

脂蛋白（ａ）［Ｌｉｐｏｐｒｏｔｅｉｎ（ａ），Ｌｐ（ａ）］是动脉粥样化中的独立危险因子，我们采用免疫组织化学技术、免疫电镜技术、酶联免疫吸附法及图像分析技术，研究了正常及不同程度动脉粥样硬化病变的尸检主动脉中Ｌｐ（ａ）分布的量及形式。结果显示：动脉粥样硬化血管壁Ｌｐ（ａ）含量显著增高，各不同病变区域Ｌｐ（ａ）有其独特的分布规律，Ｌｐ（ａ）主要位于细胞外基质中，只在少数泡沫细胞内才发现有Ｌｐ（ａ）。Ａ     

Pathogenesis of early atherosclerotic lesions in infants

High serologic lipid levels, infections, and genetic susceptibility have been proposed as possible etiologic factors of initial atherosclerotic lesions of the coronary arteries in infancy. At a recent WHO annual meeting, it was stated that breast milk substitutes cause irreparable damage in infants. This prompted us to verify whether formula feeding and parental cigarette smoking might play a role in the pathogenesis of early atherosclerotic alterations in infancy. The major epicardial coronary arteries from 36 infants dying suddenly and unexpectedly (sudden infant death syndrome) were embedded in paraffin and serially cut for histologic examination. In 67% of the cases, multifocal coronary early atherosclerotic lesions of varying entities were detected. The alterations ranged from focal plaques with mild myointimal thickening to juvenile soft plaques reducing the arterial lumen. A significant correlation was observed between the early atherosclerotic lesions and the risk factors considered. In particular, we noted different morphologic patterns related to formula feeding and cigarette smoking. Baby formula feeding and parental cigarette smoking might have an atherogenic effect on the coronary walls as from the first months of life. The lesions appear to be larger and more diffuse when both these atherogenic factors are present.     

血管树突状细胞在人主动脉粥样硬化早期病变中的分布

目的：探讨血管树突状细胞在人早期动脉粥样硬化(AS)病变中的分布模式。方法：人主动脉标本15例主要取自尸检和外科手术，常规连续切片，分别行HE及S100／CD1a免疫细胞化学染色，光镜下观察S100／CD1a阳性细胞分布情况。结果：15例HE染色标本中，2例正常，13例人动脉血管可见内膜的增厚及泡沫细胞等AS早期病理表现。9例S100／CD1a染色阳性，阳性率为69．2％。S100／CD1a阳性细胞分布在病变的内膜和外膜，外膜的S100／CD1a阳性细胞主要分布在滋养血管的周围。结论：在AS早期病变部位有血管树突状细胞的聚集，主要分布在病变血管的内膜和外膜，提示血管树突状细胞可能参与了AS早期的免疫反应。     

Intercellular contacts between stromal cells in the normal human endometrium throughout the menstrual cycle

Increasing differentiation in the human endometrium is associated with an apparent decrease in intercellular contacts between endometrial stromal cells     

Endothelial cilia in human aortic atherosclerotic lesions

Summary Primary cilia were present in the endothelial cells of human aortic fatty dots and streaks but not in those of normal intima. They had the features of cilia of the 9+0 axonemal configuration observed in many other cells. A lateral foot process and transitional fibers anchored the ciliary basal body in the cytoplasm, but rootlets were not identified in material examined. Ladder-like configurations interconnected the two centrioles (=diplosome) of control endothelium.The primary cilia of endothelium differed from those of the rudimentary type observed in smooth muscle cells in similar lesions of man, but shared many features with cilia of those present in experimental atherosclerosis in rabbit.Cilia were rarely described in vascular endothelium. It is believed that, to date, they were not reported to occur in normal or pathological arteries in man.It is being stressed that whereas the significance of these unusual organelles remains uncertain, their widespread occurrence may indicate that their role is more important than was believed previously, and they should cease being a curiosity only.Presented-in-part at the Workshop of the American Heart Association: Evolution of the Human Atherosclerotic Plaque, Rockville, Maryland, September 20–23, 1986.Dedicated to Professor Dr. Gotthard Schettler, Department of Internal Medicine, University of Heidelberg, FRG, on the occasion of his 60 birthday (April, 1987).     

Histochromatographical determination of cholesteryl ester composition in the human atherosclerotic aorta

In 14 human aortas, the cholesteryl ester (CE) composition of fatty spots was investigated histochromatographically. By a special method the intracellular lipid (foam cells) lying in the inner intima could be separated from the extracellular lipid found in the depth of the intima in many cases. The fatty acid pattern of CE of the intracellular lipid differed from that of the extracellular lipid by a relative increase in monoenoic acids (M) and trienoic acids (Tr) as well as by a relative decrease in linoleic acid (D), tetraenoic acids (Ar), and saturated fatty acids (S). In every case, the sum of the polyunsaturated fatty acids (PU) of CE (with 2 to 6 double bonds) was lower in the intracellular than in the extracellular lipid (highest difference: 23,2%, lowest difference: 5,7%). High differences of the PU-values between intra- and extracellular CE were associated with high or low D-values and with low values for Tr, Ar, and HU (high unsaturated fatty acids with more than 4 double bonds) of intracellular CE. Low differences of the PU-values between intra- and extracellular CE were accompanied with high D-values and medium or high values for Tr, Ar, and HU of intracellular CE. The results suggest that the cholesteryl esters in the intracellular lipid lose polyunsaturated fatty acids by the processes of hydrolysis and reesterification which are possibly used for other metabolic processes.     

Apolipoprotein A-1-derived amyloid in atherosclerotic plaques of the human aorta

Previous studies have shown that the amyloid localized to the aortic intima may be a biochemical entity different from other forms of localized amyloid. The amyloid fibril protein in one patient studied consisted of an N-terminal fragment of apolipoprotein A-1 (apo A-1). Since this patient was later shown to carry a missense mutation in the apo A-1 gene, leading to a deletion at position 107 of the mature protein, the question remained whether wild-type apo A-1 is amyloidogenic. In autopsy specimens from the thoracic aorta from 69 individuals, intimal atherosclerotic plaque-related amyloid was present in 11 cases (16%) and amyloid outside plaques in 37 cases (54%). The immunoreactivity of amyloid localized to the aortic intima was evaluated with the aid of antisera against N-terminal segments of apo A-1. The amyloid in association with atherosclerotic plaques was positively labelled by immunohistochemistry. The amyloid fibril protein from one patient, previously shown not to carry any mutation in the apo A-1 gene, was purified and shown by amino acid sequence analysis to be of apo A-1 nature. The result shows that wild-type apo A-1 is amyloidogenic and gives rise to a common localized form of amyloid associated with atherosclerosis.     

Composition and classification of human atherosclerotic lesions

Summary Human atherosclerotic disease can be resolved into eight types of lesion, each characterized by its composition and structure and the absence or degree of intimal injury. The eight types have been arranged in the sequence in which they may progress in complexity from the initial change in childhood or youth to the clinical endpoints in older persons. While lesions at first increase primarily by intra- and extracellular accumulation of lipid, this in itself rarely accounts for symptomatic obstruction. Lipidic lesions become symptomatic primarily by means of successively superimposed deposits of thrombotic material. Non-homogeneity of hemodynamic forces within the length of an artery account for local differences in intima thickness (adaptive intimal thickening) and, in persons with risk factors, differences in susceptibility to lesion formation. According to the degree to which they can accumulate or retain lipid and bring about secondary mechanisms, specific locations of the arterial tree have been designated asatherosclerosisresistant, atherosclerosis-prone and progression-prone.     

Composition of proteoglycans from human atherosclerotic lesions

Proteoglycans from human atherosclerotic lesions and from uninvolved aortic intima were isolated and their composition was studied. The tissues were sequentially extracted by guanidine hydrochloride followed by hydrolysis of the tissue by elastase. Chondroitin sulfate/dermatan sulfate proteoglycans were predominant in guanidine hydrochloride extracts of the tissue. Most of the heparan sulfate proteoglycans were released from the tissue by hydrolysis with elastase. The content of proteoglycan material, measured as uronate per unit weight of wet tissue, was lower in fatty streaks and fibrous plaques than in uninvolved tissue (0.58 and 0.48 mg vs. 0.7 mg/g wet tissue). The distribution of different glycosaminoglycans in guanidine hydrochloride-extracted proteoglycans was similar among the lesions and uninvolved tissue, but varied in the elastase-hydrolyzed extracts. Gel filtration studies suggested that the major proteoglycan material, chondroitin sulfate proteoglycans, from lesions had greater molecular weight than proteoglycans from uninvolved tissue. The studies indicate that alteration in intrinsic composition and molecular size of proteoglycans occurs in atherosclerotic lesions.     

Role of the monocytes and IgG in the development of atherosclerotic lesions in the aorta of rabbits

In experimental hypercholesterinemia in rabbits, immune autoradiographic studies showed that lesions in the endothelial monolayer of the aorta developed in parallel with IgG deposition in the cytoplasm of the affected endothelial cells and in different parts of atherosclerotic plaques. This process was accompanied by active replacement of blood monocytes into the internal sheath of the aorta in the zone of IgG deposition which appeared to be due to the presence in these cells of Fc-receptors for immunoglobulins. Incomplete catabolism of lipids in the cytoplasm of macrophages of monocytic genesis is one of the key events of atherosclerosis morphogenesis.