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1.
Micrococcal-nuclease digestion of rat liver nuclei selectively released mononucleosomes associated with ADP-ribosylated [Caplan, Ord & Stocken (1978) Biochem. J.174, 475-483] histone H1. Two classes of mononucleosome were detected, those that leaked out during digestion and those that were subsequently released by 5mm-sodium phosphate buffer (pH6.8)/0.2mm-NaEDTA. The former, from which histone H1 had been dissociated, contained 140-base-pair-length DNA and core histones;the latter contained core particles and mononucleosomes with histone H1 and 200-base-pair-length DNA. When normal liver nuclei were phosphorylated with [gamma-(32)P]ATP, dissociated histone H1, which could be separated from core particles with Sephadex G-200, showed (32)P uptake. (32)P uptake into histones H2A and poly(ADP-ribosyl)ated H3 was appreciable in core particles, but was less evident in nucleosomes still containing histone H1. When [(3)H]-thymidine was given to partially hepatectomized rats in S-phase, 5-10min pulses in animals of over 300g body wt. showed the presence of high-specific-radioactivity DNA in released core particles and mononucleosomes compared with DNA retained in the nuclear pellets. Mononucleosomes from rat livers in S-phase with new, [(3)H]lysine-containing histones, had higher (32)P incorporation in histones H1 and their core histones, than for di- or tri-nucleosomes. Thermal-denaturation properties of control and phosphorylated mononucleosomes and core particles were very similar; removal of histone H1 and non-histone chromosomal proteins in 0.5m-NaCl markedly increased the proportion of DNA ;melting' below 70 degrees C. 相似文献
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Transforming growth factor beta inhibits DNA synthesis in hepatocytes isolated from normal and regenerating rat liver 总被引:9,自引:0,他引:9
A J Strain A Frazer D J Hill R D Milner 《Biochemical and biophysical research communications》1987,145(1):436-442
The inhibitory action of transforming growth factor beta (TGF beta) on DNA synthesis in hepatocytes isolated from the liver of normal rats or from the liver remnant of rats 18 h following partial hepatectomy was compared. Continuous exposure to TGF beta inhibited DNA synthesis of cultured hepatocytes to a similar degree in both groups when labelled with 3H thymidine from 24-48 h or 48-72 h. At 20 pM TGF beta, 3H-thymidine incorporation was reduced by 64-78% in hepatocytes from normal liver and by 60-73% in cells from 18 h regenerating liver. The nuclear labelling index was reduced by 70-80% in all cells. Exposure to TGF beta at concentrations up to 500 pM from 0-24 h had no effect on 3H-thymidine incorporation, but exposure at 20 pM for 24 h periods thereafter was uniformally effective. These results indicate that there is no change in sensitivity of hepatocytes from 18 h regenerating liver to TGF beta, compared with normal cells, and that TGF beta may act at some point in the G1 phase of the cell cycle to inhibit hepatocyte growth. 相似文献
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Plasma membrane changes associated with rat liver regeneration 总被引:3,自引:0,他引:3
G Bruscalupi G Curatola G Lenaz S Leoni M T Mangiantini L Mazzanti S Spagnuolo A Trentalance 《Biochimica et biophysica acta》1980,597(2):263-273
The lipid composition and fluidity of plasma membranes have been studied at different stages of liver regeneration (4, 15 and 24 h after surgery). The phospholipid and fatty acid composition is not modified, whereas the cholesterol/phospholipid ratio is lower with respect to control membranes. The modification of the physical properties of the membranes has been studied directly by EPR analysis and indirectly by temperature dependence and cooperativity of some membrane-bound enzymes (Mg2+-ATPase, (Na+ + K+)-ATPase and 5'nucleotidase). Surgical operation or anaesthesia alone causes an early increase in fluidity; such an effect appears to be markedly reduced at a later stage. There seems to be a marked effect of regeneration on plasma membrane fluidity 15 h after partial hepatectomy when several parameters--surface fluidity, cholesterol/phospholipid ratio, and 5'-nucleotidase activity in the presence of concanavalin A -- are modified and indicate an increase in membrane fluidity. It is suggested that this modification of membrane properties could be related to the proliferative process. 相似文献
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Proliferation and death of hepatocytes in regenerating liver of 17-day white rat fetuses were investigated. During 2 days after liver resection (20%), animals were sacrificed every 3 h. In experimental groups, the index of Ki67-positive hepatocytes increased sharply in 15 h after liver resection. In all experimental and control groups, the ratio of the metaphase, the longest phase of mitosis, and index to mitotic index remained unchanged, indicating identical duration of hepatocytes mitoses in regenerating liver. In the regenerating and intact liver hepatocytes labeled with antibodies to caspase 3 were not detected. Thus, resection of 20% rat fetal liver did not contribute to increased apoptosis of hepatocytes. 相似文献
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Proliferation and death of hepatocytes in regenerating liver were studied in 17-day-old fetal white rats. Two days after liver resection (20%), animals were sacrificed every 3 h. In experimental groups, the index of Ki67-positive hepatocytes increased sharply 15 h after liver resection. In all experimental and control groups, the ratio of the index of the metaphase, the longest phase of mitosis, to the mitotic index remained unchanged, indicating the same duration of hepatocyte mitoses in regenerating liver. In regenerating and intact liver, hepatocytes labeled with antibodies to caspase 3 were not detected. Thus, resection of 20% fetal rat liver did not promote enhancement of apoptosis of hepatocytes. 相似文献
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A bile canalicular membrane fraction was isolated from 24-hour regenerating rat livers, and its properties were compared to those of homologous fractions prepared from the livers of sham-operated and unoperated controls. These canalicular membrane fractions were found to be closely related in terms of their morphology, their purity, their yield, and their qualitative protein banding profiles on sodium dodecyl sulfate-polyacrylamide gels. However, when a rigorous examination of plasma membrane enzyme marker activities was made, the regenerating liver membranes were shown to possess an increased specific activity of alkaline phosphatase and lower levels of Mg2+ ATPase and 5'-nucleotidase in comparison with control specific activity values. 相似文献
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After partial hepatectomy in rats, the following changes in enzymic activities were observed in the remnant liver during the prereplicative period. In the initial period of the prereplicative process, soon after removal of part of the liver, ornithine decarboxylase [EC 4.1.1.17] and IMP dehydrogenase [EC 1.2.1.14] increase. Subsequently, for entry into the S period, thymidine kinase [EC 2.7.1.75] increases simultaneously with increase in the intracellular cyclic AMP level and decrease in its phosphodiesterase [EC 3.1.4.17]. 相似文献
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Z Drahota H Rauchová V Sedlák J Koci Z Cervinková 《Physiological research / Academia Scientiarum Bohemoslovaca》1999,48(2):167-170
The increase of the membrane fluidity during the early phase of liver regeneration after partial hepatectomy was described in literature in plasma membrane and in microsomes. We found similar changes also in isolated mitochondria and in crude total membrane fraction of the liver homogenate. The administration of triiodothyronine to rats before partial hepatectomy diminished the increase of the membrane fluidity in the regenerating liver by 50%. Triiodothyronine effect is explained by hormonal modification of lipid metabolism in the regenerating liver. 相似文献
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Oriol Bachs Montserrat Soriano M. Rosa Piol Joan Serratosa Carlos Enrich 《Cell biochemistry and function》1987,5(1):37-46
We have studied calcium movement from blood into the bile by injecting 45Ca2+ intravenously and measuring the radioactivity appearing in the bile. 45Ca2+ started to appear in the bile at 3 min and maximum values were observed at 5 min after its administration. The amount of calcium secreted into the bile was proportional to the blood calcium concentration indicating that the main pathway involved in calcium movement behaved as a non-saturable system. We have also studied the 45Ca2+ circulation from blood into the bile in rats subjected to a partial hepatectomy. Thereafter, the calcium transported into the bile per gram of liver increased by about 50 per cent. Since bile flow behaved in a similar way, the biliar calcium concentration remained unmodified after hepatectomy. Determination of the activities of the Ca2+ transporting systems in isolated plasma membrane fractions from regenerating livers showed no modification in these activities suggesting that the elevation in calcium movement observed after hepatectomy is not due to an increase in the circulation of Ca2+ through the transhepatocyte pathway, an observation compatible with the absence of saturation in the transport. 相似文献
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Cell suspensions were prepared from normal and regenerating liver of adult rats by perfusion with a calcium-chelating agent (EGTA), collagenase and hyaluronidase, and the cells were incubated in culture medium. In cultures prepared from regenerating liver at 20 h after partial hepatectomy, 23 ± 4% of parenchymal cells initially incorporated [3H]TdR. This incorporation was shown to reflect semiconservative DNA replication. At least some parenchymal cells were able to complete their DNA synthesis and to progress through G2 and mitosis. Numbers of hepatocytes in mitosis increased up to 12 h of culture. On the other hand, no entry of hepatocytes into the S period was detectable in cultures prepared from normal or regenerating liver. 相似文献
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Initial RNA excess hybridization experiments employing total cell RNA and the complete complement of nonrepetitive DNA sequences showed no differences between normal and regenerating rat liver. However, when the DNA from the RNA-DNA hybrids was isolated and then reacted with homologous and heterologous RNAs the sensitivity of the assay was sufficiently improved to reveal that some of the nonrepetitive DNA transcrips present in normal liver are missing at 24 h and 48 h after a 70% partial hepatectomy. Additional experiments showed that while some of the missing sequences were common to both stages of regeneration, some were also different. The data thus suggest both quantitative and qualitative changes during liver regeneration in the population of RNA molecules transcribed from nonrepetitive DNA. 相似文献
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Yun JP Liew CT Chew EC Yin XY Lai PB Fai YH Li HK Jin ML Ding MX Li MT Lin HL Lau WY 《Journal of cellular biochemistry》2004,91(6):1269-1279
We explored the feasibility of studying nuclear matrix protein (NMP) expressions of the hepatocytes in normal and cirrhotic rat livers with liver regeneration after partial hepatectomy. Sixteen Wistar healthy rats were studied with experimental liver regeneration and/or liver cirrhosis. Two-dimensional (2-D) gel electrophoresis was used to generate these NMP compositions from these rat liver samples. Several antibodies against cytokeratin, vimentin, actin, B23, HNF4alpha, and heat shock protein 70 were used for identification by Western blot. Totally, 41 strongly stained protein spots were characterized on the 2-D gels. Thirty-four protein spots were detected in all of these rat livers, of which, cytokeratin, vimentin, actin, HNF4alpha, and heat shock protein 70 were identified. B23 was detected in the regenerated livers. Three protein spots (s33, s34, and s35) were detectable only in NMP preparation extracted from the regenerating rat livers after hepatectomy. Another three protein spots (s36, s37, and s38) were detectable only in NMP preparation extracted from thioacetamide-induced cirrhotic rat livers. Under these conditions including experimental liver regeneration and/or liver cirrhosis, Over thirty higher abundance NMPs of hepatocytes were consistently expressed and considered as common and basic NMPs. Some of the NMPs are specific for liver regeneration and may play a critical role in cell proliferation and cell cycle, and some are specific for liver cirrhosis. 相似文献
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Morphometry of normal, regenerating and cancerous hepatocytes 总被引:1,自引:0,他引:1
Z Rudzki J Szczudrawa J Stachura 《Folia histochemica et cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society》1989,27(3):141-148
During the morphometric analysis of liver cell carcinomas arising in cirrhotic livers, the sizes and shapes of 2200 cancerous and 1800 regenerative hepatocytes were measured and compared to normal hepatocytes. The neoplastic population showed significantly higher polymorphism, nucleocytoplasmic ratio and the percentage of multinucleated cells, whereas the sizes of cancerous cells were the smallest. Values for regenerative cells were mainly between those of neoplastic and normal cells. The exception was constituted by the group of very large regenerative cells which met the criteria of large liver cell dysplasia (LLCD). Low nucleocytoplasmic ratio achieved by these cells is consistent with the hypothesis of regenerative character of LLCD. 相似文献