Morphological and smooth muscle cell phenotypic changes in fetal rabbit aorta during early development

Summary Previous studies from our laboratory noted a) high aortic cholesterol esterification activity in the fetal rabbit, and b) increased susceptibility of fetal aortic explants to smooth muscle cell proliferation in culture, two features commonly associated with atherogenesis. This prompted us to examine in detail morphological development of fetal aorta and its relationship to fetal plasma cholesterol levels. Our studies made three important observations: 1) plasma cholesterol levels were high in early fetus which decreased at term; 2) in early fetus aortic endothelial cells appear to protrude into the lumen, whereas at birth the cells become flat, forming a continuous endothelium sheet; 3) in early fetus, smooth muscle cells exist predominantly in synthetic phenotype, while at birth the cells appear contractile. Despite the presence of synthetic smooth muscle cells and hypercholesterolemia in early fetal life, no accumulation of lipids was evident under transmission electron microscopy.     

Immunohistological and ultrastructural analysis of the intimal thickening in coarctation of human aorta.

OBJECTIVES: The histological nature and characteristics of aortic coarctation are not clearly defined, the aim of this study is to analyse intimal thickening in aortic coarctation. METHODS: In order to characterize the components of intimal thickening in coarctation, narrowed segments of aorta obtained after surgery from ten children were examined immunocytochemically and by electron microscopy. RESULTS: Histological analysis of aortic coarctation demonstrated a widened subendothelial region with separation of endothelial cells from the internal elastic lamina. Masson's trichrome staining showed a marked increase in extracellular matrix and cell numbers in the intimal thickening compared with normal aorta. Cellular component analysis demonstrated invagination of the intima by smooth muscle actin-positive cells, with a fragmentation of the internal elastic lamina. No proliferating smooth muscle and inflammatory cells were identified in the intima. In order to characterize the smooth muscle cell phenotypes, various smooth muscle cell markers were sought using specific monoclonal antibodies: alpha-smooth muscle actin, smooth muscle-myosin heavy chain, heavy caldesmon, desmin. In moderate coarcted aorta, at least two distinct smooth muscle phenotypes were identified. In the juxtamedial part of the intima smooth muscle, cells were differentiated and expressed all smooth muscle markers; in the subendothelial part of the intimal thickening, the majority of smooth muscle cells expressed only alpha-smooth muscle actin and appeared dedifferentiated. In regions of marked stenosis, a strong expression of smooth muscle-myosin heavy chain, and heavy caldesmon in the intimal thickening pointed to the presence of redifferentiated smooth muscle cells, not still expressing desmin. Electron microscopic examination also revealed a variety of smooth muscle cell phenotypes in the intimal thickening. In the superficial layer, smooth muscle cells appeared to be in the synthetic state, while in the deeper part, both synthetic and contractile components were identified. CONCLUSIONS: These observations indicated that human coarctation was characterized by intimal recruitment of non-proliferating smooth muscle cells with dedifferentiated phenotype. However, the presence of smooth muscle cells with an intermediate phenotype in the narrowest part of the coarctation suggest that the redifferentiation process could participate in the pathogenesis of aortic coarctation.     

Stimulation of aortic smooth muscle prostacyclin by serotonin: role of distinct receptors in contractile and synthetic states

It has been shown previously that serotonin stimulates the production of prostacyclin by bovine aortic smooth muscle cells in culture, via 5-HT2 receptors (Coughlin SR, Moskowitz MA, Antoniades HN, Levine L. Proc Natl Acad Sci USA 1981;78:7134-7138). These cells express a synthetic phenotype, whereas the majority of the smooth muscle cells in the media from adult arteries are in a contractile state. We have now compared 5-HT stimulated prostacyclin production in bovine aortic media explants, a preparation of contractile smooth muscle, with cultured smooth muscle cells derived from the explants. In the 1-10 microM range, serotonin stimulates the release of prostacyclin from the explants of bovine aortic media, cultured for a short period. In the presence of cocaine (30 microM), 1 microM was sufficient to produce a maximal effect. The stimulatory action of serotonin was sustained with time and did not induce a lasting desensitization. The effect of serotonin on the explants was inhibited only partially (+/- 30%) by ketanserin, a selective and potent 5-HT2 antagonist. It was mimicked by 5-carboxamido-tryptamine, a 5-HT1 agonist, but was only weakly inhibited by methiothepin, a 5-HT1 antagonist. As expected, in cultured smooth muscle cells, 5-carboxamido-tryptamine was only a weak agonist in stimulating prostacyclin production. In conclusion, it appears that the serotonin effect on prostacyclin production is mediated by different receptors in media explants from bovine aortic media and cultured cells obtained by outgrowth from these explants: a 5-HT2 receptor in the smooth muscle cells in culture and a receptor presenting some similarities with 5-HT1 receptors in the explants.     

Characterization of the phenotype of smooth muscle cells in human fetal aorta on the basis of ultrastructure, immunofluorescence, and the composition of cytoskeletal and cytocontractile proteins

The phenotype of smooth muscle cells (SMCs) in the aortic media of 7 human fetuses (14-20 weeks of gestation) was examined with transmission electron microscopy, immunofluorescence microscopy, and gel electrophoresis of the cytoskeletal and cytocontractile proteins. Ultrastructurally, virtually all medial cells were identified as SMCs having a poorly differentiated phenotype with a cytoplasm rich in rough endoplasmic reticulum and organelles, and with only a few myofilaments. All medial cells stained intensely with antibodies to vimentin, but only in a 20-week-old fetus could we find a few SMCs staining with antibodies to desmin. Nor was desmin detectable with SDS gel electrophoresis followed by immunoblotting, while clear bands corresponding to vimentin, myosin, and actin were present. In isoelectric focusing and two-dimensional gel electrophoresis beta-actin was the most prominent of the 3 actin isoforms in all cases. The present results show that SMCs in the media of fetal human aorta have a poorly differentiated phenotype, which morphologically and biochemically resembles that previously described in the aorta of fetal and newborn rat, in the arterial intima after endothelial injury, in atherosclerotic lesions, and after spontaneous modulation of medial SMCs in culture.     

Three-dimensional architecture of elastic tissue in athero-arteriosclerotic lesions of the rat aorta

The overall three-dimensional architecture of elastic tissue in athero-arteriosclerotic lesions of the ascending aorta of the rat was studied, using scanning electron microscopy (SEM), after hot-formic acid extraction followed by a freeze-drying method. The lesions were induced by feeding the rats a diet containing 2% cholesterol for 6 weeks and administering massive doses of vitamin D2 for the first 4 days. SEM revealed the synthesis of elastins mainly in the intima and the degradation in the internal elastic lamina (IEL) and medial concentric elastic lamellae. Intimal elastic elements involved in atherosclerotic lesions were composed of an intricately tangled network of fibrous elastins or a row of elastic laminae that surrounded many rectangular compartments, which were separated by fenestrated septum-like elastins and were assumed to be the sites of smooth-muscle-cell proliferation. These observations indicated remodelling of the elastic laminae in the intima. Degradation in the IEL and media was attributed to either vitamin D2 intoxication, which was characterized by the formation of a large degraded portion, or an enhanced elastolytic process. These alterations of elastic elements probably reflect the exact frame of the modes of migration of aortic smooth muscle cells in the development of athero-arteriosclerotic lesions, thereby indicating the role of smooth muscle cells in the progress of atherosclerosis.     

Modulation of human aorta smooth muscle cell phenotype: a study of muscle-specific variants of vinculin, caldesmon, and actin expression.

Vinculin- and caldesmon-immunoreactive forms and actin isoform patterns were studied in samples of normal and atherosclerotic human aorta. After removal of adventitia and endothelium, the remaining tissue was divided into three layers: media, muscular-elastic (adjacent to media) intima, and subendothelial (juxtaluminal) intima. In media of normal aorta, meta-vinculin accounted for 41.0 +/- 0.9% (mean +/- SEM) of total immunoreactive vinculin (meta-vinculin + vinculin); 150-kDa caldesmon accounted for 78.2 +/- 5.1% of immunoreactive caldesmon (150-kDa + 70-kDa); the fractional contents of alpha-smooth muscle actin, beta-nonmuscle, and gamma-isoactins were 49.0 +/- 0.6%, 30.4 +/- 0.6%, and 20.8 +/- 0.8%, respectively. Muscular-elastic intima was very similar to media by these criteria. In subendothelial intima, the fractional content of meta-vinculin and 150-kDa caldesmon was significantly lower (6.9 +/- 1.5% and 32.7 +/- 7.0%, respectively) than in muscular-elastic intima and media, whereas the isoactin pattern was identical to that in adjacent layers, demonstrating the smooth muscle origin of subendothelial intima cells. In atherosclerotic fibrous plaque, the fractional content of alpha-actin was decreased in subendothelial intima, rather than in media and muscular-elastic intima. Additionally, the proportion of subendothelial intima cells [i.e., the cells that express low amounts of smooth muscle phenotype markers (meta-vinculin, 150-kDa caldesmon, and alpha-actin)] in the total intima cell population increased dramatically in atherosclerotic fibrous plaque. The results suggest that changes in the relative content of meta-vinculin and 150-kDa caldesmon as well as alpha-actin in human aortic intima are associated with atherosclerosis although, in subendothelial intima of normal aorta, a certain smooth muscle cell population exists that expresses reduced amounts of "contractile" phenotype markers, even in the absence of the disease.     

大鼠胸主动脉平滑肌细胞的培养与鉴定

目的探讨平滑肌细胞培养方法,了解生长特性。方法用组织贴块法培养大鼠胸主动脉平滑肌细胞,用相差显微镜观察其生长情况,用免疫荧光染色分析其抗原的表达。结果培养3日可见组织块周边有细胞长出,2周达亚融合,传代后细胞“谷峰”生长明显。SM-α-肌动蛋白和Calponin抗体免疫荧光染色呈阳性。结论组织贴块法是简单、经济、高效的平滑肌细胞培养方法,为心血管疾病发病机制的研究提供了理想的细胞模型。     

Rhythmic structures of the aorta in children and young adults

517 samples of the aorta (descending thoracic and abdominal) from children aged 0 to 14 years and 70 men aged 20-29 years were investigated. Rhythmic structures (RS) were found in 4% of still-borns and in children in their first days of life. Thereafter, the occurrence of RS systematically increased with age, and after the first year of life they occurred in 60% of cases. In men aged 20-29 years RS were transformed in 40% into ribbon-shaped fibrous plaques (FP); in cases when RS were absent, small FP occurred only in 7% of cases. The initial phase of RS development is characterized by impaired and disorganized elastic fibres at the border between the intima and the media. This is followed by migration of smooth muscle cells from the media into the intima and intimal thickening, accompanied with neoformation of irregularly arranged elastic fibres. The next phase, is characterized by fibrous transformation of RS into FP. Similar to aortic RS coronary arteries of children develop "cushions" as a result of hyperplasia of intimal elastic fibres and smooth muscle cells.     

Changes in morphology of elastin fibers during development of the tunica intima of monkey aorta

Summary The normal development of elastin fibers in the thoracic aorta was studied in fetal, young, and adult monkeys. Tissue was examined by scanning electron microscopy (SEM) after NaOH treatment and by transmission electron microscopy (TEM). The NaOH treatment of fixed tissues effectively removed collagen fibers and enabled three-dimensional visualization of the elastin fibers. In intact fetal aortae, the internal elastic lamina (IEL) was situated immediately beneath the endothelium. This IEL consisted of superficial, longitudinally arranged bundles of elastin fibrils and an underlying solid sheet containing round fenestrations. In neonates, diffuse intimal thickening was observed. In the young and young-adult monkeys, the aortae exhibited intimal thickening with slender but split IEL. One of the most important findings of this study was that elastin fibers in the intimal thickening, as well as smooth muscle cells, ran in a longitudinal fashion. This was in contrast with the elastic laminae of the media which were mainly oriented circumferentially. Subendothelial elastin fibers in this intimal thickening combined with longitudinally arranged microfibrils which formed close associations with endothelial stress fibers. In some adult monkey aortae with well-developed intimal thickening, a complex meshwork of slender elastin fibers was also found beneath the endothelium. The development of the intimal elastin fibers is discussed in relation to hemodynamic forces.     

Endothelial healing following defined injury to rabbit aorta. Depth of injury and mode of repair

A study has been made of the healing of a narrow deep injury to rabbit aortic endothelium which also involves damage to the media of the vessel. The injury was produced using a nylon catheter containing a wire filament; the injury was approximately 150 micron in width and damaged up to 3 elastic lamellae. Immediately after injury platelet aggregates were observed over the injured areas, several hours later large numbers of leukocytes were also seen to adhere. Two days after injury a non-thrombogenic neointimal surface was observed over deeply injured areas; endothelial cells could be identified covering the injured area at 6 days. The healing process following the injury has been directly compared with the healing of rabbit aortic endothelium following a superficial injury of similar width, where endothelial cells are removed without significant damage to the media of the vessel [1]. The results show that (a) following a narrow injury to the aorta which causes damage to the media platelet aggregation and proliferation of smooth muscle cells occurs, (b) despite the disruption of subendothelial components, endothelium rapidly regenerates over the narrow injured area, although not as quickly as for a superficial injury.     

What controls smooth muscle phenotype?

Enzyme-dispersed smooth muscle cells from the adult pig aortic media in the first few days of primary culture are in the contractile phenotype and do not divide when challenged with 5% WBS. After 6–8 days the isolated cells spontaneously undergo a change in phenotype where contraction cannot be stimulated and the cells respond to mitogens in WBS by logarithmic growth. The change in phenotype is reversible if the cells are seeded sufficiently densely (5 X 10⁴ to 1 X 10⁵/ml) that a confluent monolayer results after less than 1 week of proliferation, but is irreversible if the cells are seeded sparsely (1 X 10³ to 5 X 10³ /ml) and take more than 2 weeks of proliferation to reach confluence. When the cells are seeded so densely (10⁶/ml) that a confluent monolayer is present from day 1, the cells do not undergo a change in phenotype but remain indefmitely in the contractile state.

The spontaneous modulation of phenotype of isolated smooth muscle cells can be inhibited by a confluent monolayer of contractile smooth muscle or endothelial cells in co-culture with the sparsely seeded smooth muscle such that the two cell layers are not in contact but bathed by the same nutrient medium. Smooth muscle modulation can also be inhibited by a factor extracted from pig and rabbit aortic tissue, and its effects mimicked by commercially available sodium heparin at 50 units/ml.

The results are discussed in relation to control of proliferation of arterial smooth muscle as in the formation of atherosclerotic plaques.     

The urokinase receptor mediates basic fibroblast growth factor-dependent smooth muscle cell migration through baboon aortic explants

The urokinase receptor is required for vascular smooth muscle cell migration in vitro, but may not be needed in vivo since smooth muscle cell migration and intimal hyperplasia after arterial injury in mice are not affected by urokinase receptor gene deletion. We have used baboon aortic explants as a bridge between cell culture and in vivo experiments to determine if the urokinase receptor is required for smooth muscle cell proliferation and smooth muscle cell migration in primate vessels. Levels of urokinase receptor in explants increased with time after explantation, while blockade of urokinase receptor with an antibody decreased smooth muscle cell proliferation and smooth muscle cell migration from the explants. A blocking antibody to basic fibroblast growth decreased levels of urokinase and urokinase receptor in explants, and it decreased smooth muscle cell migration and mitogenesis. These results suggest that the factor urokinase receptor plays a positive role in smooth muscle cell migration and proliferation in injured primate arterial tissue, in part mediating the pro-migratory and proliferative effects of basic fibroblast growth factor released by damaged smooth muscle cells.     

Effect of Saireitoh on rabbit smooth muscle cell proliferation and experimental atherosclerosis

Saireitoh is a traditional Chinese medicine that is often given to patients with nephrotic syndrome or glomerulonephritis. Studies have reported that Saireitoh stimulates intrinsic steroid secretion in rats and suppresses the proliferation of fibroblasts in vitro. We examined the effects of Saireitoh on vascular smooth muscle cell proliferation and migration in vitro and experimental atherosclerosis in vivo. Saireitoh rabbit serum obtained from New Zealand White rabbits which were given a diet containing 2% Saireitoh for 3 days significantly inhibited [3H]-thymidine incorporation by smooth muscle cells, which were isolated from thoracic aorta explants of rabbits. The addition of 10% Saireitoh rabbit serum to a culture medium containing smooth muscle cells inhibited DNA synthesis by 50% as compared with a control culture to which 10% normal rabbit serum was added. We also found that the number of smooth muscle cells in the culture containing Saireitoh rabbit serum was decreased. When PDGF was used as a chemoattractant, we demonstrated that Saireitoh rabbit serum slightly inhibits the migration of smooth muscle cells. In in vivo experiments, Saireitoh did not suppress the development of atherosclerosis but tended to reduce the damage. We concluded that although Saireitoh inhibited the proliferation of smooth muscle cells, the effect of prevention on the development of atherosclerosis is weak in the in vivo condition.     

Development of the aorta in the chick embryo: structural and ultrastructural study

A structural and ultrastructural study was designed to analyze systematically the cellular events which take place in the aortic wall between days 7 and 21 of chick embryo development. Between days 7 and 18, increase in total diameter, number of cell layers, and aortic wall thickness are highly correlated, whereas between days 18 and 21 the total diameter increase is correlated mainly with an increase in vessel lumen diameter. Cell layers of smooth muscle cells showing an immature or synthetic phenotype arise from progressive association and organization of mesenchymal cells originated from an endothelial activation process in which a hyaluronic acid-rich extracellular matrix seems to be involved. It is suggested that the process of endothelial activation takes place between days 7 and 18 of embryonic development provided that within that period the typical cellular events which are involved in such a process take place (hypertrophy, reorientation, invagination, mitotic activity, acquisition of migratory appendages, endothelial detachment and incorporation into adjacent spaces). This endothelial activation has been recognized as a selective multiphasic process required for the transition of endothelial cells into mesenchyma.     

Cytoskeletal features of rat aortic cells during development. An electron microscopic, immunohistochemical, and biochemical study

Actin, vimentin, desmin, and tropomyosin distribution in rat aortic endothelial and smooth muscle cells has been studied during development using fetal (18 to 20 days of gestation), and 5- and 14-day-, and 5-, and 12-week-old rats. Endothelial cells of newborn animals actively replicate and contain many actin stress fibers, whereas, in adult animals, replication is minimal and actin stress fibers are rare. The actin, vimentin, desmin, and tropomyosin content of smooth muscle cells increases gradually from fetal to adult animals. The number of desmin-containing cells also increases from 13% in fetal rats to 51% in adult rats. The beta-actin isoform is predominant in fetal and newborn animals, but gradually the alpha-isoform becomes quantitatively the most important, as seen by bidimensional polyacrylamide gels. Several analogies exist between the features of developing smooth muscle and what is known for developing striated muscle cells. The evolution of cytoskeletal features from fetal to adult animals is remarkably the opposite of what takes place in: (1) rat aortic smooth muscle cells proliferating after an endothelial injury, (2) human arterial smooth muscle cells present in atheromas, and (3) actively growing rat aortic smooth muscle cells in vitro. Thus, the assumption that pathological or cultured smooth muscle cells are "dedifferentiated" is supported by our biochemical observations.     

Phenotype related changes of intimal smooth muscle cells from human aorta in primary culture.

To study the functional characteristics of smooth muscle cell (SMC) phenotypes, we have investigated myosin expression, cell proliferation, collagen production and low-density lipoprotein (LDL) receptor activity in intimal SMCs of normal human aorta during their growth in primary culture. By staining with rabbit antibodies to smooth muscle myosin (ASMM) 3 cell types could be distinguished in culture: homogeneously stained cells, cells with discontinuous myosin fibrils and myosin-negative cells. The ratio of cell types greatly changed with culture growth: on days 5, 7 and 14 it was 82:1:17%, 70:5:25% and 10:30:60%, respectively. After 5-6 days of culture intimal SMCs began to proliferate and DNA-synthesizing nuclei were seen 1.5-4.3 times more frequently in myosin-negative cells than in cells with homogeneous myosin distribution. At that time the number of cells reacted with monoclonal antibody (MAb) to an epitope shared collagen types I and III started to increase. By double immunofluorescence staining it was shown that the cultured cells containing both ASMM and MAb markers were found 2.0-4.8 times more rarely than MAb-positive staining in myosin-negative cells. During the first 5 days in culture LDL binding and uptake were diminished in intimal cells with intercellular lipid inclusions independently of their myosin staining pattern, but their activity increased with culture growth. Thus, SMCs from human aortic intima change their phenotype on days 6 and 7 in primary culture as manifested by alteration of myosin expression, increased cell proliferation, collagen production and LDL receptor activity. Changes in myosin expression, however, are not an essential prerequisite for cell proliferation and collagen production.     

Identification and expression of scavenger receptor SR-BI in endothelial cells and smooth muscle cells of rat aorta in vitro and in vivo

In this study, we used immunoelectron microscopy to investigate the subcellular localization of scavenger receptor class B type I (SR-BI) in the arterial walls of rats. The expression of SR-BI in cultured endothelial and smooth muscle cells of rat aorta after exposure to high-density lipoprotein (HDL) was also investigated by immunofluorescence microscopy and immunoblotting analysis. A peptide containing residues 495-509 from mouse SR-BI (mSR-BI) plus an NH2-terminal cysteine was coupled to hemocyanin to generate mSR-BI antiserum in rabbits. Reactivity of antiserum against the synthetic peptides was confirmed with an enzyme-linked immunosorbent assay (ELISA). The results showed that SR-BI was specifically localized on the surface of the endothelial cells and smooth muscle cells. SR-BI was also observed in the cytoplasm of smooth muscle cells. Immunoblotting analysis indicated that SR-BI was expressed in the cell membrane. The levels of SR-BI increased gradually from 1 to 3 h and decreased at 24 and 48 h after cholesterol-loaded cells were incubated in the culture medium containing HDL. We conclude that SR-BI, a functional receptor for HDL, is expressed in the aortic endothelial cells as well as in smooth muscle cells. This receptor also responds to the presence of HDL in the culture medium.     

Nov gene encodes adhesion factor for vascular smooth muscle cells and is dynamically regulated in response to vascular injury

Nephroblastoma overexpressed (NOV) is a member of the CCN family (connective tissue growth factor, CYR61, and NOV) of proteins that are involved in regulating the proliferation, differentiation, and adhesion of a variety of cell types. We have examined the expression of the NOV: gene and NOV protein by vascular smooth muscle cells (VSMCs), in vitro and in vivo, and the effects of recombinant NOV on VSMCs. Rat aortic VSMCs were found to express NOV: mRNA and NOV protein in vitro and in vivo. NOV: expression in adult rat tissues was very high in the aorta and was detected only weakly in the brain and lung by Northern analysis (relative levels 33:3:1). During postnatal development (3 days to 12 weeks), the expression of NOV: was correlated with markers of the differentiated smooth muscle cell phenotype (smooth muscle myosin heavy chain and SM22 alpha). In the rat carotid artery balloon injury model, NOV: was detectable by in situ hybridization and was downregulated in the media of the injured artery compared with the uninjured artery at 7 and 14 days after injury. Expression in the developing intima was barely detectable at 7 days after injury except for strong expression at the luminal surface. At 14 days after injury, NOV: expression was substantially increased throughout the intima. In vitro studies of the function of NOV protein showed that it promoted VSMC adhesion via a mechanism that was divalent cation and Arg-Gly-Asp independent but that it did not modulate VSMC proliferation or phenotype. The strong expression and dynamic regulation of NOV: in the arterial wall, together with its ability to promote VSMC adhesion, suggest that it may be involved in homeostasis and repair.     

Ascending aorta dilatation in aortic valve disease: morphological analysis of medial changes

We investigated whether and how the severity of medial degeneration lesions varies along the circumference of the dilated intrapericardial aorta. Two groups of aortic wall specimens, respectively harvested in the convexity and concavity of ascending aorta in 72 patients undergoing surgery for dilatation of the intrapericardial aorta associated with aortic valve disease, were separately sent for pathology, morphometry, and ultrastructural examination. Cystic medial necrosis, fibrosis, and elastic fiber fragmentation were classified into three degrees of severity; their mean degree and morphometric findings in the convexity and in the concavity specimens were compared by paired t-test. Correlation between echocardiographic degree of aortic dilatation and severity of medial degeneration was assessed separately for each of the two groups of specimens. Morphologically, medial degeneration was found in all cases; a higher mean degree was found in the convexity group (2.39 ± 0.58 vs 1.44 ± 0.65 in the concavity group; P < 0.001). At morphometry normal smooth muscle cells in the convexity specimens were significantly reduced (P = 0.007); the length (P = 0.012) and number (P = 0.009) of elastic fibers reduced and increased, respectively. Moreover, in the convexity specimens a significantly smaller amount of smooth muscle cells and an increase of immunohistochemical labeling of apoptosis-associated proteins in the subintimal layer of the media was noticed. Correlation between aortic ratio and medial degeneration degree was significant in the convexity group (P < 0.001), but not in the concavity group (P = 0.249). Scanning electron microscopy analysis confirmed morphological results and allowed us to better distinguish the early pathological cavities from the microvessels, which were in the outer media in normal aorta and ubiquitous in aortitis or atherosclerosis. Electron transmission microscopy analysis showed changes in the extracellular matrix and smooth muscle cells, and these changes increased from the intima to the adventitial layer of the media. In dilated intrapericardial aorta, medial degeneration changes and expression of apoptosis-associated proteins are more marked in the ascending aorta convexity, likely due to hemodynamic stress asymmetry. Ultrastructural findings allow us to distinguish the early medial changes not yet evident on light microscopy.     

Angiotensin II type 1 receptor antagonist downregulates nonmuscle myosin heavy chains in spontaneously hypertensive rat aorta

The aim of this study was to clarify the differences between the angiotensin II type 1 (AT1) receptor antagonist and the angiotensin-converting enzyme (ACE) inhibitor on smooth muscle and nonmuscle myosin heavy chain isoforms in aortic smooth muscle cells of Wistar-Kyoto rats and spontaneously hypertensive rats. All 4 myosin heavy chain isoforms are heterogeneously expressed in the smooth muscle cells of the aortic tunica media in 20-week-old rats, and the contractile-type myosin heavy chains are highly expressed in smooth muscle cells of the aortic tunica media compared with the synthetic-type myosin heavy chains. Both the AT1 receptor antagonist and the ACE inhibitor had the same effects on hemodynamics, smooth muscle cell hypertrophy and proliferation, fibrosis, and vascular remodeling in spontaneously hypertensive rats. However, the AT1 receptor antagonist had a more potent effect on the downregulation of the synthetic-type myosin heavy chains than the ACE inhibitor in spontaneously hypertensive rat aortic tunica media. In contrast, these effects of the AT1 receptor antagonist and the ACE inhibitor on hemodynamics, morphology, fibrosis, and expression of myosin heavy chain isoforms in smooth muscle cells of the aortic tunica media were not observed in Wistar-Kyoto rats. Thus, within 6 weeks, the AT1 receptor antagonist might modulate the cellular composition of myosin heavy chain isoforms in smooth muscle cells more efficiently than the ACE inhibitor, without morphological changes in the spontaneously hypertensive rat aorta.