Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.24.3 (collagenase)
 18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Subendothelial cells (SEC) were obtained from the inner intimal layer of adult human aorta by collagenase treatment. SEC were identified in primary culture either as smooth muscle cells by staining with FITC-labeled antisera against human smooth muscle myosin or as macrophages, foam cells and contaminating endothelial cells by their uptake of malondialdehyde treated low density lipoproteins labeled with fluorescent dye 3,3'-dioctadecylindocarbocyanine. Between 1 and 5 days in culture, along with smooth muscle cells (SMC, 38-82%), endothelial cells (0-9%), macrophages and foam cells (2-32%), one more type of cell was found. This cell type resembled SMC in size and shape, but was not stained by antisera to SMC myosin. By ultrastructural criteria these cells were characterized as modulated SMC for they contained prominent rough endoplastic reticulum and Golgi complex together with basement membrane and a large number of plasmalemmal vesicles. Like SMC they reacted with phalloidin and were stained by anti-vimentin but not by anti-desmin monoclonal antibodies. The proportion of such cells varied from 5 to 33% of total cell number and increased in parallel to macrophages and foam cells in vessels with well developed atherosclerotic lesions. We conclude that the applied technique may be used for identification of cultured vascular cells including modulated SMC.
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PMID:Identification of intimal subendothelial cells from human aorta in primary culture. 313 80

Organ-derived endothelia have been shown to exhibit distinct patterns of morphology and growth responsiveness in vitro. This report describes the development, cloning and establishment of long-term serial cultures of rat vascular endothelial cells derived from cerebrocortical resistance vessels (small arteries and arterioles). Modification of our previous published technique for establishing resistance vessel-derived smooth muscle cells (RV-SMC) resulted in enhanced levels of endothelial outgrowth from collagenase-treated microvessel fragments. Although primary culture growth consisted predominantly of SMC, subsequent subcultivation of these cultures revealed the presence of distinct endothelial cell clusters within the SMC monolayer. Serial cloning of these isolates resulted in a homogeneous population of cells with the characteristic endothelial cobblestone growth pattern and positive immunofluorescence for factor VIII-related antigen. Previously established RV-SMC frozen stocks provided an additional source for obtaining resistance vessel endothelial cells. This was made possible by the slow proliferation rate of early-passage RV-SMC and their inability to withstand freezing procedures. Endothelial cells from both preparations were identical and designated resistance vessel derived endothelial cells RV-EC. Upon long-term cultivation (> P15), confluent RV-EC cultures expressed spontaneous multicellular cord development that stained positive for factor VIII-related antigen. Cell growth studies demonstrated that RV-EC were capable of significant growth when maintained in serum-free conditions. Growth kinetics using serum-free conditioned medium demonstrated mitogenic activity indicating the presence of an autocrine growth factor. Increase growth responsiveness was also noted in RV-EC when treated with a variety of peptide growth factors. These results indicate that resistance vessel endothelium can be successfully isolated and maintained in long-term serial cultures. Furthermore, the availability of cultured EC and SMC from this unique microvascular site will enable examination of cerebrovascular endothelial-smooth muscle cell interactions in vitro and may help to elucidate the mechanisms of altered vascular function in disease states.
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PMID:Isolation and characterization of cerebral resistance vessel endothelium in culture. 814 May 79

Migration to the intima and other responses of M-SMC in the rat carotid artery and abdominal aorta after balloon injury were investigated in vivo. Migration occurred intensively between the second and fifth days after injury. About 80% of the cells were in the G1 and S phases of the cell cycle. The majority of the migrating cells were therefore simultaneously proliferating. Positive values of 42.3%, 48.9%, 44.4%, and 32.8% of the migrating cells on the fifth day in the carotid artery for PDGF-B, elastase III B, MMP-I, and MMP-9, were observed, respectively. Many of the cells expressed messages of PDGF-A and elastases II and III B by in situ hybridization. Fine structures of the migrating cells were characterized as a synthetic phenotype of the smooth muscle cell with reduced attachment to their surrounding ECM. A biphasic proliferative response of the M-SMC appeared on the second and fifth days. Migration occurred correspondingly in the proliferative period. The populations of M-SMC positive in immunostainings for PDGFs, their receptors, elastase III B, and MMP-1 and MMP-9 also increased biphasically, around 12 h and five days after the injury. The results of these studies suggest that the migrating cells were proliferative and synthesizing PDGFs, elastases, and collagenases.
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PMID:Migration of medial smooth muscle cells to the intima after balloon injury. 918 23

The matrix metalloproteinases are a large group of proteases with a central role of the degradation of all types of extracellular matrix. The present study investigated expression of matrix metalloproteinases (MMP-1, -2, -9) and their inhibitors (TIMP-1, -2) in chronic Aneurysm of the Thoracic Aorta (ATA) and Post-Stenotic Dilatation of the ascending aorta due to valvular aortic stenosis (PSD). Fragments of the ascending aorta that had been taken from the patients during coronary by-pass surgery were used as controls. Immunohistochemical investigation showed that medical SMC in the samples taken from aortas with ATA and PDS expressed a stronger immunoreactivity for MMP-1, -2, -9 and TIMP-1, -2 as compared to controls. It can be suggested that during formation of ATA and PSD, production of MMPs and TIMPS by medial smooth muscle cells is of great importance.
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PMID:[Morphogenesis of thoracic aorta aneurysms: investigation of matrix metalloproteinases and their tissue inhibitors]. 1247 14

Chlorotyrosine is an oxidative product of hypochlorous acid and l-tyrosine, and is considered as a biomarker for oxidative stress and cardiovascular disease. However, it is not clear whether chlorotyrosine could directly contribute to vascular pathogenesis. In this study, we investigated the effect and potential mechanisms of chlorotyrosine on human aortic smooth muscle cell (AoSMC) migration. With Boyden chamber and wound healing assays, chlorotyrosine significantly increased AoSMC migration in a concentration- and time-dependent manner. In addition, chlorotyrosine significantly increased the expression of several key molecules related to cell migration including PDGF receptor-B (PDGFR-B), matrix metalloproteinases (MMP-1 and MMP-2) and integrins (alpha3, alphaV, and beta3) in AoSMC at both mRNA and protein levels. Furthermore, chlorotyrosine also increased superoxide anion generation in AoSMC with the fluorescent dye dihydroethidium (DHE) staining. Activation of mitogen-activated protein kinases (MAPKs) was analyzed with Bio-Plex Luminex immunoassay and Western blotting. Chlorotyrosine induced a transient phosphorylation of ERK1/2, but not JNK and p38 MAPKs. Antioxidants including selenomethionine (SeMet) and Mn(III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) as well as ERK1/2 inhibitor PD98059 effectively blocked chlorotyrosine-induced AoSMC migration. Thus, these findings demonstrate new biological functions of chlorotyrosine in human SMC migration, which may play a crucial role in the vascular lesion formation.
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PMID:Chlorotyrosine promotes human aortic smooth muscle cell migration through increasing superoxide anion production and ERK1/2 activation. 1828 Oct 51