Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.27.1 (RNase)
 16,360 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

1. Activated hepatic lipocytes are central to the pathogenesis of liver fibrosis as the principal source of both interstitial collagens and matrix-degrading metalloproteinases. In progressive fibrosis there is a failure to degrade interstitial collagens with a reported decrease in collagenase activity. In these studies we investigate expression of the potent collagenase inhibitor, tissue inhibitor of metalloproteinase-1, and interstitial collagenase in end-stage autoimmune chronic active hepatitis and activated human hepatic lipocytes in culture. 2. Messenger RNA transcripts for interstitial collagenase and tissue inhibitor of metalloproteinase-1 in explanted human liver were quantified by ribonuclease protection assay and densitometric analysis. This indicated that tissue inhibitor of metalloproteinase-1 and interstitial collagenase expression in autoimmune chronic active hepatitis were also coordinately up-regulated. 3. Using Northern analysis of RNA from human lipocytes in primary culture on plastic, mRNA for interstitial collagenase could not be detected in unstimulated cells but was present after stimulation with tumour necrosis factor alpha. Tissue inhibitor of metalloproteinase-1 mRNA was present in unstimulated lipocytes and up-regulated fivefold in response to tumour necrosis factor alpha. Using activity assay of serum-free conditioned media, interstitial collagenase could not be detected in unstimulated primary cultures, primary cultures stimulated with tumour necrosis factor alpha or transforming growth factor beta-1 (n = 3 and n = 4 respectively) or in passaged lipocytes (n = 6). In contrast, free tissue inhibitor of metalloproteinase-1 activity was present in unstimulated and passaged cultures and this was increased in response to tumour necrosis factor alpha and transforming growth factor beta-1.(ABSTRACT TRUNCATED AT 250 WORDS)
 ...
PMID:Tissue inhibitor of metalloproteinase-I and interstitial collagenase expression in autoimmune chronic active hepatitis and activated human hepatic lipocytes. 767 71

Liver fibrosis results from a relative imbalance between synthesis and degradation of matrix proteins. We have previously described release of the protein collagenase inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), by culture-activated human hepatic stellate cells (HSCs). In this study, we have investigated the relative expression of TIMP-1 and interstitial collagenase in culture-activated rat HSCs and rat models of liver injury and fibrosis. The complementary DNA (cDNA) for rat TIMP-1 was obtained by homology polymerase chain reaction (PCR) and sequenced. By Northern analysis using this probe, TIMP-1 messenger RNA (mRNA) expression was up-regulated with HSC activation by culture on plastic as defined by cellular expression of procollagen-1. Interstitial collagenase mRNA was expressed in early 1. Interstitial collagenase mRNA was expressed in early culture (<4 days) but became undetectable in more activated cells (7-21 days). By activity assay of serum-free cell-conditioned media, TIMP-1 was found to be released in increasingly concentrations with duration of culture on plastic. Expression of TIMP-1 interstitial collagenase, and procollagen-1 mRNAs were studied in rat models of biliary and parenchymal injury (bile duct ligation and CC14 administration) by ribonuclease protein assay. TIMP-1 mRNA expression was increased at 6, 24 hours, and 3 days after bile duct ligation and was also shown to rise in acute CC14 liver injury and remain elevated as the liver became fibrotic. TIMP-1 expression preceded procollagen-1 expression in both models. In contrasts, interstitial collagenase mRNA levels remained similar to control values throughout both models of liver injury. Total cellular RNA from hepatocytes, HSCs, and kupffer cells freshly isolated from livers after acute CC14 injury was subjected to Northern analysis. TIMP-1 transcripts were observed in nonparenchymal cells only. We suggest that increased expression of TIMP-1 relative to interstitial collagenase by HSCs may promote progression of liver fibrosis in these rat models by preventing degradation of secreted collagens.
 ...
PMID:Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. 870 59

Interstitial collagenase is secreted by the osteoblast in response to bone-resorbing agents. Previously, we cloned the rat interstitial collagenase cDNA from UMR 106-01 rat osteoblastic osteosarcoma cells. We demonstrated that induction of collagenase by PTH, a powerful resorbing agent, in UMR 106-01 cells is in part transcriptional. In the present study we isolate and characterize the rat interstitial collagenase gene. The gene consists of 10 exons and spans approximately 12 kbp. The major transcriptional start site, determined by primer extension analysis and confirmed by RNase protection assay, is 25 nucleotides upstream of the translational start site. The previously isolated cDNA was missing the 5'-untranslated sequence in addition to 17 nucleotides of the signal sequence of the preproenzyme; therefore, we also present these data. Chloramphenicol acetyl transferase (CAT) analyses were performed on the 5'-upstream region of the gene. These data indicate that PTH appears to mediate its effect through an AP-1 consensus-binding sequence (-51). Footprint analysis demonstrates protein binding to this site. Site-specific mutagenesis markedly decreased protein binding, which correlated directly with a decrease in CAT activation by PTH. Supershift data indicate that cAMP response element binding protein (CREB) is binding to this AP-1 consensus sequence. In addition we demonstrate that PTH induces phosphorylation of CREB.
 ...
PMID:Parathyroid hormone induction of rat interstitial collagenase mRNA in osteosarcoma cells is mediated through an AP-1-binding site. 881 27

We compared two models of cardiac fibrosis in which collagen synthesis is controlled at different levels. Regulation is pretranslational in aldosterone-salt-induced hypertension in young rats and posttranslational in 24-month-old rats. However, little is known about the role of matrix metalloproteinases (MMP) in fibrosis development. Ventricular MMP activities were studied by zymography, and MMP-2 and MMP-1 mRNA levels were determined using slot-blot and ribonuclease protection assay, respectively. After 1 month of aldosterone-salt treatment, proMMP-2, MMP-2, and proMMP-1 collagenolytic activities and their gene expression were unchanged compared with sham-operated rats. After 2 months, total MMP-2 activity was increased by 40% with parallel stimulation of its gene expression. These changes were localized by in situ zymography within the media of coronary vessels. These results suggest that MMP play a prominent role in vascular remodeling during the first steps of hypertension. During aging, however, there were 40% and 45% decreases in MMP-2 and proMMP-1 activity, respectively, with a corresponding down-regulation of MMP-2 mRNA. These observations suggest that depression of the degradative pathway is partly responsible for age-associated fibrosis. Thus, MMP have differing involvements in the cardiac remodeling associated with hypertension or aging.
 ...
PMID:Differential regulation of matrix metalloproteinases associated with aging and hypertension in the rat heart. 916 91

Delayed reperfusion has a beneficial effect on prognosis, independent of infarct size. One potential mechanism to explain this observation may be an effect on infarct healing. In this study, the impact of delayed reperfusion on two aspects of the healing process was examined, the activity of matrix metalloproteinase (MMP) enzymes and the expression of fibronectin (FN) mRNA. The rat model of coronary artery ligation was used and rats were randomly assigned to delayed reperfusion (150 min following coronary ligation) or permanent ligation. Animals were subsequently killed 1, 2, 3 and 7 days following infarction. Infarct tissue was harvested for MMP activity (zymography), FN mRNA (RNase protection analysis) and protein (immunofluorescence microscopy and Western analysis), and collagen content (hydroxyproline concentration). Infarction produced marked activation of MMP-1, -2, and -9. Reperfusion significantly attenuated the activity of these enzymes (approximately 50% reduction in MMP-1, P=0.03 and ;60% reduction in MMP-2 at 7 days, P=0.001; approximately 55% reduction in MMP-9 at 24 h and 84% reduction at 48 h, P=0.01 and 0.002, respectively). Delayed reperfusion also produced a trend toward a greater increase in FN mRNA 24 h following infarction and immunofluorescent staining suggested the presence of more FN protein at this point. These data demonstrate that delayed reperfusion alters matrix metalloproteinase activity and fibronectin mRNA expression in the infarct zone. The impact of these changes on infarct healing and their association with the improved prognosis of a patent infarct vessel following infarction will require further study.
 ...
PMID:Delayed reperfusion alters matrix metalloproteinase activity and fibronectin mRNA expression in the infarct zone of the ligated rat heart. 929 68

There is compelling in vitro and in vivo evidence to implicate mast cells in the development of fibrosis. However, an important question remains as to the mechanisms by which mast cells mediate fibrosis. Recent evidence from our laboratory (Gruber et al., 1997, J. Immunol. , 158:2310-2317) has revealed that tryptase, the unique and abundant serine protease of human mast cells, is capable of activating fibroblasts by stimulating chemotaxis, proliferation, and procollagen mRNA synthesis. Regulation of matrix metalloproteinase (MMP) expression is another key step in connective tissue remodeling. Therefore, the effect of tryptase on fibroblast MMP expression was investigated. Proteolytically active tryptase did not alter the cellular mRNA levels for fibroblast MMP-1, MMP-2, MMP-3, and MMP-9 as detected by RNase protection assays. Moreover, tryptase did not alter the basal levels of MMP-1, MMP-2, MMP-3, MMP-9, or the tissue inhibitor of MMP-1 (TIMP-1) in fibroblast conditioned media as detected by specific enzyme-linked immunosorbent assay (ELISA). These results indicate that tryptase does not increase MMP expression in normal dermal fibroblasts. Moreover, these data strengthen the potential role of this unique serine protease as a potent fibrogenic factor.
 ...
PMID:Mast cell tryptase does not alter matrix metalloproteinase expression in human dermal fibroblasts: further evidence that proteolytically-active tryptase is a potent fibrogenic factor. 1049 10

The purpose of this study was to determine whether the production of interstitial collagenase mRNA in response to parathyroid hormone (PTH) changes with osteoblast phenotypic development. To accomplish this, cells derived from fetal rat calvaria were examined. The calvarial osteoblasts, which proliferate when placed in culture, can be made to differentiate after confluence. Studies were performed on cells while they were proliferating, at confluence, and during the differentiation process. The cells were treated with PTH for various times, and interstitial collagenase mRNA was quantified by RNase protection assay. We concluded that the ability of PTH to induce interstitial collagenase mRNA in these cells increased with osteoblast phenotypic development. We also determined that the response could be mimicked by combining the effect of 8-bromo-cAMP and 12-O-tetradecanoyl-phorbol-13-acetate, stimulators of the protein kinase A and protein kinase C pathways, respectively, both known to be activated by PTH. The binding of nuclear factors to two regions previously reported to be important for PTH induction of the gene in UMR 106-01 cells was also examined. These data indicated that the binding of nuclear factors to oligonucleotides encompassing the TRE (-51) or the PEA3 (-80) elements changed with development of the osteoblast phenotype. The latter was also shown to be PTH responsive.
 ...
PMID:Induction of rat interstitial collagenase (MMP-13) mRNA in a development-dependent manner by parathyroid hormone in osteoblastic cells. 1096 42

Components of the extracellular matrix take part in tissue rebuilding as well as activating surface-bound growth factors. In the present study, expression and selected activities of urokinase-type plasminogen activator (uPA), matrix metalloproteinases (MMPs), their inhibitors (plasminogen activator inhibitor 1 (PAI-1) and tissue inhibitors of metalloproteinases (TIMPs)) were examined in bovine oviducts by RT--PCR, ribonuclease protection assay and activity assays. A high content of mRNA encoding for uPA was detected before ovulation with a three-fold decrease after ovulation. In contrast, PAI-1 expression appeared to be stable during the oestrous cycle. Oviductal flushings produced caseinolytic zones in zymograms containing plasminogen at approximately 50 kDa and 28 kDa. An activity assay for uPA showed highest net activity during the early to mid-luteal phase. Increased TIMP-1 and MMP-2 mRNA concentrations were found around the time of ovulation compared with the luteal phase. In contrast, MMP-1 mRNA transcripts were enriched during the early to mid-luteal phase. Gelatin zymograms detected a 70--72 kDa protease activity showing an oestrous cycle-dependent activity with highest activity before ovulation. Reverse zymography detecting TIMPs revealed proteins between 21 kDa and 24 kDa. Only for the smallest (21 kDa) protein were amounts increased around the time of ovulation compared with the luteal phase. The observation that several extracellular matrix components were regulated distinctly in bovine oviducts indicates that local interactions between these components, growth factors, gametes and the embryo are possible and may influence fertilization and early embryonic development.
 ...
PMID:Differential expression of extracellular matrix components in the bovine oviduct during the oestrous cycle. 1142 36

The abnormal expression of matrix metalloproteinases (MMPs) plays an important role in the invasion of malignant gliomas into the surrounding normal brain tissue. This study showed that curcumin has broad-spectrum inhibitory activity against MMP gene expression in human astroglioma cells. RNase protection assay showed that curcumin inhibited the PMA-induced mRNA expression of MMP-1, -3, -9, and -14. Curcumin repressed the DNA binding and transcriptional activities of AP-1, which is a common upstream modulator of MMP-1, -3, and -9 gene expression. In addition, curcumin suppressed the PMA-induced MAP kinase activities, which were differentially involved in modulating the MMPs. This suggests that the inhibition of MMP transcriptions by curcumin is mediated at least in part through the AP-1 and MAP kinase pathways. Curcumin was also found to significantly repress the in vitro invasion of glioma cells. Therefore, the broad-spectrum inhibition of MMP gene expression by curcumin might provide a novel therapeutic strategy for treating gliomas.
 ...
PMID:Curcumin is a potent broad spectrum inhibitor of matrix metalloproteinase gene expression in human astroglioma cells. 1619 11