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1.
Summary The distribution of actin, myosin and tropomyosin in freshly isolated and short-term cultured rat hepatocytes was investigated by use of both rhodaminyl-phalloidin staining and immunofluorescence techniques. The cytoskeletal proteins were mainly located in distinct areas of the hepatocyte membrane, corresponding to their accumulation in the bile-canalicular region of liver tissue. In freshly prepared cells, these sections resembled sharp, angled or branched bands, similar to the pattern of hemicanaliculi. During incubation in a monolayer culture, these bands were transformed to circular formations. Simultaneously, enclosed bile-canalicular spaces between undissociated hepatocytes were visualized by staining of actin, myosin, and tropomyosin. The preservation of canalicular cytoskeletal structures in isolated hepatocytes is an indication of cellular polarity. Our findings suggest a uniform association of membrane-bound F-actin with myosin and tropomyosin. 相似文献
2.
用不同自由基源处理的胆红素与鼠肝细胞相互作用,结果表明:胆红素自由基可引起鼠肝细胞脂质过氧化,使总谷胱甘肽及GSSG水平明显下降,细胞损伤后,乳酸脱氢酶外漏,上述结果与自由基浓度正相关,而与其种类无关,牛血清白蛋白有明显的抑制作用。差示光谱表明:胆红素可能与细胞色素P-450快速形成络合物。根据以上结果,重点讨论了胆红素自由基对肝细胞损伤的化学本质及其与胆结石形成的关系。 相似文献
3.
臧宁 《中国生物化学与分子生物学报》1994,10(2):191-195
用大鼠肝脏门静脉或肝静脉周围的肝细胞来研究葡萄糖和酮体生成的区域分布。肝细胞通过毛地黄皂苷-胶原酶灌流技术分离。门静脉周围肝细胞的γ谷氨酰转肽酶的活性比肝静脉周围肝细胞高2.4倍;而谷氨酰胺合成酶的活性则相反,肝静脉周围肝细胞高出56倍。门静脉周围肝细胞的内源性葡萄糖合成比肝静脉周围肝细胞高1.57倍。给予刺激葡萄糖异生的底物,门静脉周围肝细胞的葡萄糖合成则增加1.7-2.1倍。肝静脉周围肝细胞的内源性酮体生成比门静脉周围肝细胞高1.3倍。给予能明显刺激酮体生成的辛酸盐,肝静脉周围肝细胞的酮体生成仅略为增加。我们的结果证实,在基础和刺激的条件下,葡萄糖的异生在门静脉周围肝细胞中优先,而酮体生成仅在肝静脉周围肝细胞占微弱的优势。 相似文献
4.
Toshitsugu Nakamura Masao Hotchi 《Virchows Archiv. B, Cell pathology including molecular pathology》1993,63(1):11-16
DNA strand breaks (nicks) in non-parenchymal cells (NPCs) in CCl4-induced acute or chronic liver injury in rats were detected using an in situ nick translation method; their dynamic changes
were analysed in relation to the proliferation pattern of hepatocytes and NPCs, as revealed by bromodeoxyuridine (BrdU)-up-take.
In acute injury, hepatocyte proliferation started before centrilobular necrosis had occurred, whereas BrdU-labeled sinusoidal
NPCs markedly increased only after centrilobular necrosis was apparent. DNA breakages in NPCs paralleled the proliferation
pattern of these cells, suggesting that nicks are physiological, and reflect proliferation and activated gene expression.
In chronic injury, liver cirrhosis developed after 9 weeks, but BrdU-labeling of hepatocytes was almost the same level as
that in untreated liver. The number of BrdU-labeled NPCs showed only a slight increase, while those with DNA breakages were
much more frequent in the cirrhotic stage, suggesting a significant role for NPCs in the fibrotic process. These results indicate
that DNA strand breaks in NPCs act as a marker for activation states such as proliferation, differentiation and/or activated
gene expression. 相似文献
5.
应用[~3H]TdR掺入离体培养大鼠肝细胞DNA的方法,测定由本室提取的pHSS的生物活性。结果表明,pHSS可显著促进原代培养大鼠肝细胞的DNA合成,其促进率约为对照组的10倍左右。培养液中血清浓度对pHSS的生物活性表达有显著影响,不同浓度血清可以使pHSS表现出不同的量效关系,这些结果在Buffello大鼠肝细胞系的实验中得到进一步证实。在低剂量pHSS的刺激下,不同年龄大鼠肝细胞的[~3H]TdR掺入率无显著差异。但高剂量时,pHSS对幼鼠作用不明显。 相似文献
6.
Phosphorylation of a 16-kDa protein by diacylglycerol-activated protein kinase C in vitro and by vasopressin in intact hepatocytes 总被引:1,自引:0,他引:1
A replication-defective Simian virus 40 genome, with a deletion of about 120 nucleotides in the region encoding the N-terminal fourth of the large T antigen, has been isolated from the DNA of Simian cells transformed by SV40. Both the original transformants, and the murine transformants obtained by transfection with this cloned mutant DNA, produced a large T antigen displaying in immunofluorescence an exclusively cytoplasmic localization. The protein apparent molecular mass (83 kDa) was about 6% smaller than that of normal karyophilic large T. Restriction analysis showed that the deletion eliminated two close HinfI sites, at nucleotides 4459 and 4376 (map unit 0.50). 相似文献
7.
S. Ferri 《Protoplasma》1980,103(1):99-103
Summary Freshwater teleost hepatocytes show hypertrophied Golgi complexes 3 days after a single intraperitoneal injection of cadmium chloride (10 mg/Kg). 相似文献
8.
We examined the interaction of glycolipid-containing phospholipid vesicles with rat hepatocytes in vitro. Incorporation of either or the monosialoganglioside, GM1, enhanced liposomal lipid uptake 4–5-fold as judged by the uptake of radioactive phosphatidylcholine as a vesicle marker. Cerebroside enhanced phospholipid uptake only when incorporated into dimyristoyl, but not into egg phosphatidylcholine vesicles. The lack of cerebroside effect in egg phosphatidylcholine-containing vesicles appeared to be due to a limited exposure of the carbohydrate part of the glycolipid as suggested by the reduced agglutinability of those vesicles by Ricinus communis agglutinin.In contrast to the results with radioactive phosphatidylcholine, we observed only a 20% increase in vesicle-cell association as a result of glycolipid incorporation, when a trace amount of [14C]cholesteryloleate served as a marker of the liposomal lipids or when using the fluorescent dye, carboxyfluorescein, as a marker of the aqueous space of the vesicles. By the same token, intracellular delivery of vesicle-contents was only slightly enhanced (approx. 10%).The discrepancy between the association with the cells of phosphatidylcholine on the one hand and cholesteryoleate or entrapped marker on the other suggests different mechanisms of uptake for these markers. Our results are compatible with the notion that the main effect of incorporation of glycolipids into the vesicles is the enhancement of exchange or transfer of phospholipid molecules between vesicles and cells. Incubation of the cells with galactose or lactose, prior to addition of vesicles, suggests that this enhanced phospholipid exchange or transfer involves specific recognition of the terminal galactose residues of the glycolipid vesicles by a receptor present on the plasma membranes of hepatocytes. 相似文献
9.
José Remacle 《生物化学与生物物理学报:生物膜》1980,597(3):564-576
The in vitro incorporation of cytochrome into purified plasma membranes was investigated by biochemical and immunological methods. Plasma membrane preparations incorporated three times less cytochrome than did microsomal preparations; 60% of this cytochrome could not be reduced by the NADH-cytochrome reductase and was considered as being bound to the plasma membrane. The morphological observations made after the immunochemical labeling of cytochrome clearly showed a good but asymmetrical distribution of the ferritin labeling: only the inner face of the plasma membrane incorporated cytochrome . These results are discussed with respect to theories which concern the subcellular membrane relationships in the cell. 相似文献
10.
George A. Cook Vincent H. Gattone Andrew P. Evan Robert A. Harris 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1983,763(4):356-367
The structural changes accompanying digitonin-induced release of enzymes and metabolites from isolated hepatocytes have been studied by scanning and transmission electron microscopy. In the initial phase, characterized by total release of the cytosolic marker enzyme, lactate dehydrogenase, the plasma membrane was immediately damaged, rapidly followed by extensive damage to the endoplasmic reticulum. The shape of the cell, however, was maintained, and the mitochondria and nucleus remained tightly held together by the cytoskeleton. Mitochondria remained intact initially, whereas the cytosol became less electron dense and the nuclear chromatin was more dispersed. An intermediate phase was characterized by total release of adenylate kinase and most of the glucose-6-phosphatase, marker enzymes for the mitochondrial intermembrane space and the endoplasmic reticulum, respectively. The outer mitochondrial membrane was ruptured, but mitochondria maintained their normal matrix electron density. In the final phase, characterized by the beginning of citrate synthase release from the mitochondrial matrix space, the mitochondria became swollen, and only the nucleus, inner and outer mitochondrial membranes, and the cytoskeleton could be clearly distinguished. Although the plasma membrane could not be readily discerned in electron micrographs after the initial phase, the plasma membrane marker enzyme 5′-nucleotidase remained associated with digitonin-treated hepatocytes. Acetyl-CoA carboxylase was released much more slowly than lactate dehydrogenase, indicating some severe restriction on its release. The release of acetyl-CoA carboxylase closely paralleled the release of glucose-6-phosphatase. The controlled exposure of hepatocytes to digitonin, therefore, leads to the sequential release of soluble, compartmentalized cellular components and some membrane-bound components, but the mitochondrial membrane, cytoskeleton and the nucleoskeleton survive even long-term digitonin treatment. 相似文献