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

The presence and distribution of collagenase in experimental CCl4 cirrhosis of the liver in rats has been studied by immunohistochemical techniques. A monospecific anti-rat uterus collagenase antibody was raised in rabbits and used for indirect immunofluorescence staining of liver sections obtained from rats in both the reversible and irreversible stages of CCl4-induced cirrhosis. Collagenase is present assoicated with connective tissue septums as long as cirrhosis is reversible, and it is not detectable in the irreversible stage. In animals sacrificed during the transition between the reversible and irreversible stages of cirrhosis, collagenase appeared bound to the outer surfaces of connective tissue septums and was absent from the deeper portions. These observation suggest that the irreversibility of experimental CCl4 cirrhosis of the liver is associated with a disturbance in the mechanisms of collagen degradation, which may be a deficiency in collagenase activity, a change in the susceptibility of the substrate, or a combination of both factors.
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PMID:Collagenase in experimental carbon tetrachloride cirrhosis of the liver. 20 92

Previous investigators have reported a protective effect of some prostaglandins and of the prostaglandin E2 analogue enprostil on carbon tetrachloride (CCl4)-induced injury of liver cells. In the present study liver cells were isolated from the rat liver by collagenase perfusion and suspended in F-10 medium, containing 20% foetal bovine serum, 1% gentamicin, and 1% glutamine. In the first study cells were cultured in T-flasks with 3 ml suspension of 6 x 10(6) cells/ml, and in the second study (extended dose response) cells were cultured in tissue culture wells with 0.5 ml cell suspension. Misoprostol was added to groups of cultures 15 min before CCl4, 2 microliters/ml, and the number of living cells was counted 45 min after the first addition. The number of living cells was compared with those of other groups with CCl4 only and control groups. In the first experiment misoprostol was given in doses of 200, 400, and 800 ng/ml medium and in the second experiment in 0.1, 1, 10, 100, and 1000 ng/ml medium. CCl4 is an agent well known to be toxic to liver cells, and in cultures to which only CCl4 was added, the number of living cells was significantly reduced compared with controls. When 0.1 ng misoprostol was added before CCl4, no significant difference in the number of living cells was shown compared with cultures with CCl4 only. On the other hand, misoprostol given in doses from 1 ng to 1000 ng before CCl4 resulted in a higher number of living cells, indicating a protective effect.
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PMID:Effect of the prostaglandin E1 analogue misoprostol on the carbon tetrachloride-induced injury of rat liver cells in culture. 194 73

In order to explore the cellular source(s) and the behaviour of the collagenolytic activity previously described in rat liver homogenates, in the reversibility of experimental cirrhosis of the liver, enriched suspensions of hepatocytes and of sinusoidal liver cells were obtained by a procedure which employs low EDTA concentrations and no bacterial collagenase. Cell suspensions were prepared from three different groups of animals: 1) normal controls, 2) rats with CCl4-induced cirrhosis of the liver, and 3) rats with swine serum-induced cirrhosis of the liver. Animals were sacrificed in each group upon completion of treatment and also after 3, 6 and 12 months. In each liver wet weight and collagen concentration were determined, and collagenolytic activity of both enriched cell suspensions was measured separately. In addition, histological studies of liver tissue and ultrastructural examination of cell suspensions were performed by standard procedures. Enriched suspensions of both normal hepatocytes and sinusoidal liver cells display Ca2(+)-dependent collagenolytic activities. Both cell suspensions obtained from each of the two types of cirrhotic livers show normal or slightly increased average levels of collagenase activity at the time of treatment discontinuation, when average liver collagen content ranges from 6 to 10-fold over normal, suggesting that the normal collagenase/collagen ratio is disturbed and that collagenolytic activity is deeply decreased in relation to the actual liver collagen load.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Collagenase of hepatocytes and sinusoidal liver cells in the reversibility of experimental cirrhosis of the liver. 198 May 58

Circulating AC levels as well as antibodies against AC-protein adducts are increased in non-alcoholic liver injury. To identify the adducts, we used rats with CCl4-induced cirrhosis. Liver subcellular fractions were analyzed by immunochemical staining of protein slot blots and of electrophoretically separated proteins, transferred to nitrocellulose, using AC-protein adduct-specific antibodies. One reactive protein of about 200 kD was detected in the liver soluble fraction and in the cytosol of isolated hepatocytes and, to a lesser extent in the liver microsomes of CCl4-treated rats; in control animals, this reactivity was much weaker. The immunopositive AC adduct co-migrated with the beta 1,2 dimer of rat collagen type I; it was sensitive to digestion by a highly purified collagenase and also reacted with anti-rat collagen type I-specific IgG. In addition, comparison of peptides of the CNBr-digested, immunoprecipitated AC adduct with those of rat collagen type I revealed a high degree of similarity. Thus, AC adduct formation occurs in liver injury of non-alcoholic origin, and a target protein appears to be related to collagen type I, most likely the procollagen precursor.
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PMID:Acetaldehyde-collagen adducts in CCl4-induced liver injury in rats. 217 75

Previous experiments showed that the presence of high levels of acute phase reactants (APR) enhance CCl4-induced liver fibrosis in the rat. A high correlation was found between the degree of fibrosis and alpha 2-macroglobulin of the rat (alpha 2-macrofetoprotein, alpha M-FP) used for monitoring the acute phase response. This acute phase reaction was provoked by epinephrine just before CCl4 treatment was started. In the present study we analyzed the effect of APR by repeating these experiments and estimating liver neutral collagenase with a synthetic substrate and endogenous collagen as a substrate, and liver prolyl-4-hydroxylase. A strong depression of liver collagenase activity was found in rats with a preceding acute phase reaction contrary to the rats that underwent CCl4 treatment only. A high level of alpha M-FP correlated negatively with collagenase activity. Also in vitro alpha M-FP proved to inhibit collagenase activity. Prolyl-4-hydroxylase was increased in the rats during acute phase reaction and correlated highly and positively with alpha M-FP, haptoglobin, and ceruloplasmin. Thus high levels of APR promote development of CCl4-induced fibrosis, partly by anticollagenase activity and partly because of enhancement of prolyl-4-hydroxylase activity. The latter phenomenon can also be explained by the presence of APR, but this has to be proved.
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PMID:Mechanisms by which acute phase proteins enhance development of liver fibrosis: effects on collagenase and prolyl-4-hydroxylase activity in the rat liver. 242 60

A short-term in vivo method for assay of repair and replication of rat liver DNA has been developed, by which possible hepatocarcinogens could be identified in a few days. F344 rats were treated orally with two genotoxic hepatocarcinogens, dimethylnitrosamine (DMN) and 2-acetylaminofluorene (2AAF), or a nongenotoxic hepatocarcinogen, carbon tetrachloride (CCl4). Then at suitable times after treatment, their hepatocytes were isolated by a two-step collagenase perfusion technique in situ and incubated with [3H]dThd with or without hydroxyurea, which inhibits DNA replication. Their nuclear DNA was then extracted and the incorporation of [3H]dThd into nuclear DNA was determined in a liquid scintillation counter. Unscheduled DNA synthesis (DNA repair), induced by DMN at doses of 2.5-10 mg/kg body weight and by 2AAF at doses of 12.5-50 mg/kg body weight, could be detected 2 h and 4 h after their administration as an increase of DNA synthesis of up to 5.8-fold and 6.0-fold, respectively, in the presence of hydroxyurea. Replicative DNA synthesis, induced by CCl4 at a dose of 200 mg/kg body weight, could be detected 48 h after its administration as a 23-fold increase of DNA synthesis in the absence of hydroxyurea and was inhibited approximately 97%-99% by hydroxyurea. Replicative DNA synthesis induced by 2AAF at a dose of 25 mg/kg body weight 16 h after its administration could be detected as a 6.8-fold increase of DNA synthesis in the absence of hydroxyurea. These results show that unscheduled and replicative DNA synthesis can be clearly distinguished by simultaneous measurements of the incorporation of [3H]dThd into nuclear DNA in the presence and absence of hydroxyurea.
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PMID:In vivo short-term assays of repair and replication of rat liver DNA. 276 99

Prostaglandins have been reported to reduce the carbon tetrachloride (CCl4)-induced liver cell injury in rats. The object of the present experiments was to examine the effect of the prostaglandin E2 analogue enprostil on the survival of isolated liver cells exposed to CCl4. Liver parenchymal cells were isolated from rat livers by collagenase perfusion and released into a 'suspension' buffer. Aliquots of the cell suspension were incubated with 1 micrograms or 0.5 microgram CCl4, and to parallel test suspensions 20 ng enprostil was added 5-10 min before CCl4. Incubation was performed on ice, at room temperature, and at 37 degrees C. The average percentage of dead cells after CCl4 treatment was significantly reduced by pretreatment with enprostil at room temperature (1 microgram CCl4: 69 +/- 21% and 44 +/- 13%, respectively) and after 10 min of incubation at 37 degrees C (1 microgram Cl4: 56 +/- 25% and 37 +/- 27%; 0.5 microgram CCl4: 51 +/- 33% and 29 +/- 18%, respectively). When the liver cell mortality approximated 100% after long-term incubation at 37 degrees C, no protective effect of enprostil was observed.
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PMID:Prostaglandins and CCl4-induced liver cell mortality. The effect of the prostaglandin analogue enprostil. 314 36

The interaction between fat-storing cells (FSCs) and Kupffer cells (KCs) in vitro has been studied in an attempt to clarify certain aspects of the pathogenesis of fibrotic process in the liver. FSCs and KCs were isolated from the livers of rats either treated with CCl4 for 6 weeks, or with vitamin A for 6 weeks or from untreated rats by the pronase-collagenase digestion method. FSCs were further purified by centrifugation over a double layered metrizamide gradient, and KCs were separated from other sinusoidal cells by the dish adherence technique. FSCs from CCl4-treated rats divided rapidly, while those from vitamin A-treated rats divided slowly, as compared with untreated rats. Furthermore, the proliferation of FSCs was enhanced in the presence of KCs from CCl4-treated rats, but was slightly suppressed by KCs from normal and vitamin A-treated rats. This enhancement was mediated by a non-dialyzable, soluble factor present in the conditioned medium of KCs from CCl4-treated rats, but was not detected in the conditioned medium of KCs from normal or vitamin A-treated rats. From the present study, a growth factor secreted by KCs from CCl4-treated rats may play an important role in controlling the proliferation of FSCs during the pathogenesis of liver fibrosis.
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PMID:Kupffer cells from CCl4-induced fibrotic livers stimulate proliferation of fat-storing cells. 355 40

In vitro studies were carried out on isolated rat hepatocytes to examine further the proposed cytoprotective actions of prostaglandins (PG) using carbon tetrachloride (CCl4) as the toxic agent. Isolated hepatocytes, prepared by collagenase, were cultured in Leibowitz-15 medium. Following preincubation, CCl4 (300 or 150 micrograms/ml) was added to the hepatocytes. Treatment with Indomethacin (INDO), 16, 16-dimethyl-PGE2(PGE2) and prostacyclin (PGI2) was assayed in the cultures. Cell damage was measured by lactic dehydrogenase (LDH) release. 6-keto-PGF1 alpha was measured in the supernatant by direct radioimmunoassay. The results showed PGI2 (30.0 ng/ml) treatment 30 min after CCl4 (300 micrograms/ml) addition to be highly protective (p less than 0.001 versus CCl4 control). PGE2 (3 ng/ml) showed similar protection (p less than 0.001). INDO (2 micrograms/ml) following CCl4 (150 micrograms/ml) demonstrated increased cell death (p less than 0.001). INDO (0.5 micrograms/ml) reduced 6-keto-PGF1 alpha production (p less than 0.05). Low dose ethanol (1.5 micrograms/ml) increased 6-keto-PGF1 alpha production (p less than 0.05). Ethanol (1.5 micrograms/ml), added to stimulate endogenous PG production, was cytoprotective when added prior to CCl4 (p less than 0.01). This protection was suppressed by INDO. Ethanol added after CCl4 was not protective. We conclude that exogenously added PGI2 and PGE2 are cytoprotective in this in vitro model and that endogenous PG production may play a protective role in the initial stages of cellular damage.
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PMID:Cytoprotective effect of prostaglandins on isolated rat liver cells. 388 50

16,16-Dimethyl PGE2 (dmPGE2) has previously been shown to protect the in vivo rat liver against CCl4-induced damage. These studies were undertaken to determine if this protection could be demonstrated in vitro where factors of absorption, secretion, and blood flow are not present. Primary hepatocyte cultures were established by perfusing rat liver with collagenase. Hepatocytes were plated at a density of 2 X 10(4) cells/cm, allowed 90 min to attach, then stabilized in L15 medium for 18 h. Hepatocytes were then challenged with CCl4 with concomitant exposure to 10(-9) to 10(-5) M dmPGE2, stearic acid, oleic acid, or ethanol vehicle (0.00001 to 0.1%). After 1 h, challenge was aspirated and cells were stained with 0.04% trypan blue to determine viability. Hepatocytes in the vehicle groups took up more trypan when exposed to CCl4 than those treated with dmPGE2, stearic acid, or oleic acid at concentrations of 10(-9) to 10(-7) M. At 0.1% ethanol vehicle protected as well as all other treatments. Protection against CCl4 by dmPGE2, stearic, and oleic acids as well as high concentrations of ethanol may occur by altering the metabolism of CCl4.
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PMID:16,16-Dimethyl PGE2 and fatty acids protect hepatocytes against CCl4-induced damage. 403 Jun 26


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