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1.
The substrate specificity and subcellular location of the major sialidases of three types of rat blood cells were characterized and compared with those of the known three types of rat liver sialidase, which have been designated intralysosomal, cytosolic, and plasma membrane-associated sialidases. Platelets and leucocytes contain mainly an acid sialidase, which is highly active towards oligosaccharides and 4MU-NeuAc, and erythrocytes possess a high level of a sialidase acting on gangliosides. A Percoll gradient centrifugation study showed that the former is located in lysosomes and the latter in plasma membrane. When the sialidase was solubilized and partially purified from erythrocyte ghosts, the enzyme was found to hydrolyze actively gangliosides but only poorly other substrates such as 4MU-NeuAc, oligosaccharides, and glycoproteins. The sialidase partially purified from rat liver membrane fraction exhibited the same substrate specificity. It is concluded that the major sialidase of platelets and leucocytes corresponds to hepatic intralysosomal sialidase while erythrocytes contain almost exclusively a ganglioside sialidase which corresponds to hepatic plasma membrane sialidase. 相似文献
2.
Intact rat or human erythrocytes and their isolated (ghost) membranes were incubated with the high speed supernatant fraction of homogenates derived from 32P-labeled rat livers. Phospholipid molecules were transferred between the red cell membranes and the liver extracts, as reflected by the convergence of their specific radioactivities with time. Whereas ghosts usually approached isotopic equilibrium with the liver supernatant fraction during a few hours of incubation at 37° C, the exchange of phospholipids by intact cells was no more than one-half, even after 18 hr. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and sphingomyelin were all exchanged in both intact cells and ghosts, albeit to different extents. (A control experiment, incubating 32P-labeled rat erythrocytes or ghosts with unlabeled rat liver extracts, also demonstrated the exchange of all four major phospholipids.) These data may signify that the phospholipids on the cytoplasmic side of the membrane of intact erythrocytes do not exchange with the phospholipids in exogenous liver extracts. If so, all four major phospholipid classes would appear to be present to some extent at both membrane surfaces. The first inference is in agreement with several other studies on this membrane, while the second inference is not. 相似文献
3.
Hubert Kolb Amelie Kriese Victoria Kolb-Bachofen Hans-Albert Kolb 《Cellular immunology》1978,40(2):457-462
Neuraminidase-treated rat lymphocytes adhere strongly to rat hepatocytes in vitro. Binding between cells is due to stereo-specific interactions between a mammalian hepatic membrane lectin and galactosyl residues which are exposed on the lymphocyte surface after removal of sialic acid residues. The hepatic galactose specific lectin may play a role in the trapping of recirculating desialylated lymphocytes in the liver. 相似文献
4.
J. Schlepper-Schfer N. Holl V. Kolb-Bachofen E. Friedrich H. Kolb 《Biology of the cell / under the auspices of the European Cell Biology Organization》1985,52(3):253-258
The mechanism by which macrophages recognize tumor cells is still unknown. We have studied interactions between rat liver macrophages and rat L 5222 leukemia cells. These tumor cells, but not normal leukocytes or erythrocytes, adhere to freshly isolated macrophages in vitro. Binding of tumor cells by macrophages can be inhibited by N-acetyl-D-galactosamine, D-galactose and more potently by glycoproteins with terminal N-acetyl-D-galactosamine or D-galactose residues. Tumor cell adhesion is calcium-dependent. The relevant leukemia cell membrane structures which bear terminal beta-D-galactosyl or related residues have been determined as trypsin- and pronase-sensitive, and hence may presumably be glycoproteins. The tumor cell receptor on liver macrophages appears to be a lectin with the carbohydrate specificity N-acetyl-D-galactosamine greater than D-galactose greater than L-fucose. 相似文献
5.
J Schlepper-Sch?fer N Holl V Kolb-Bachofen E Friedrich H Kolb 《Biology of the cell / under the auspices of the European Cell Biology Organization》1984,52(3):253-258
The mechanism by which macrophages recognize tumor cells is still unknown. We have studied interactions between rat liver macrophages and rat L 5222 leukemia cells. These tumor cells, but not normal leukocytes or erythrocytes, adhere to freshly isolated macrophages in vitro. Binding of tumor cells by macrophages can be inhibited by N-acetyl-D-galactosamine, D-galactose and more potently by glycoproteins with terminal N-acetyl-D-galactosamine or D-galactose residues. Tumor cell adhesion is calcium-dependent. The relevant leukemia cell membrane structures which bear terminal beta-D-galactosyl or related residues have been determined as trypsin- and pronase-sensitive, and hence may presumably be glycoproteins. The tumor cell receptor on liver macrophages appears to be a lectin with the carbohydrate specificity N-acetyl-D-galactosamine greater than D-galactose greater than L-fucose. 相似文献
6.
Angelo Benedetti Alessandro F. Casini Marco Ferrali Mario Comporti 《The Biochemical journal》1979,180(2):303-312
The effects on cellular structures of products of peroxidation of rat liver microsomal lipids were investigated. A system containing actively peroxidizing liver microsomal fraction was separated from a revealing or target system by a dialysis membrane. The target system, contained in the dialysis tube, consisted of either intact cells (erythrocytes) or subcellular fractions (liver microsomal fraction). When liver microsomal fractions were incubated with NADPH (or an NADPH-generating system), lipid peroxidation, as measured by the amount of malonaldehyde formed, occurred very rapidly. The malon-aldehyde concentration tended to equilibrate across the dialysis membrane. When the target system consisted of erythrocytes, haemolysis occurred abruptly after a lag phase. The lysis was greatly accelerated when erythrocytes from vitamin E-deficient rats were used, but no haemolysis was observed when erythrocytes from vitamin E-treated rats were used. When, in the same system, freshly prepared liver microsomal fractions were exposed to diffusible factors produced by lipid peroxidation, the glucose 6-phosphatase activity markedly decreased. A similar decrease in glucose 6-phosphatase activity, as well as a smaller but significant decrease in cytochrome P-450, was observed when the target microsomal fractions were exposed to diffusible factors derived from the peroxidation of liver microsomal lipids in a separate preincubation step. These and additional experiments indicated that the toxicological activity is relatively stable. Experiments in which the hepatic microsomal fractions destined for lipid peroxidation contained radioactively labelled arachidonic acid, previously incorporated into the membranes, showed that part of the radioactivity released from the microsomal fraction into the incubation medium entered the dialysis tube and was recovered bound to the constituents of the microsomal fractions of the target system. These results indicate that during the course of the peroxidation of liver microsomal lipids toxic products are formed that are able to induce pathological effects at distant loci. 相似文献
7.
Binding and uptake studies of in vitro aged or senescent rat erythrocytes by isolated rat liver macrophages suggest recognition by galactose-specific receptors on the cell surface of the macrophages. We analyzed carbohydrates exposed on old erythrocytes by plant lectins in an agglutination assay in comparison with freshly isolated untreated erythrocytes. Rat erythrocytes aged in vitro by storage are agglutinated by a panel of lectins that do not react with freshly isolated erythrocytes. Specificity of agglutination was shown by inhibition with monosaccharides. Antibodies eluted from senescent rat erythrocytes agglutinate in vitro aged as well as senescent rat erythrocytes, but not freshly isolated cells nor human erythrocytes. Galactose-specific lectins isolated from rat liver give similar results; they also agglutinate normal human erythrocytes. Agglutination by the liver lectin is inhibitable by galactose and N-acetylgalactosamine but not by N-acetylglucosamine or mannose. Furthermore, rat liver macrophages devoid of galactose-specific receptors show markedly reduced binding of senescent rat erythrocytes. We conclude that recognition of old rat erythrocytes is mediated by two systems: old erythrocytes expose different terminal sugar residues or a different arrangement of glycans when compared to young erythrocytes, rendering them recognizable by liver lectins and by autoantibodies. 相似文献
8.
L Gattegno F Fabia D Bladier P Cornillot 《Biomedicine / [publiée pour l'A.A.I.C.I.G.]》1979,30(4):194-199
Evidence is presented to indicate a generalized role for the terminal sialic acid residues of circulating erythrocytes. After reinjection into their donors, neuraminidase-treated human, rabbit, rat and dog erythrocytes were promptly removed from the circulation : intect erythrocytes, previously incubated under the same conditions but without neuraminidase, were removed after a significantly longer period. The neuraminidase-treated erythrocytes were cleared by the liver and in a little part by the spleen. Old and young human, rabbit, rat erythrocytes contained different quantities of stromal sialic acid, significantly lowered on the old cells. But the half-life of old intact rabbit erythrocytes is sigificantly shorter than that of neuraminidase-treated young erythrocytes with a similar minidase-treated young erythrocytes with a similar sialic acid content. Indeed sialic acid is not the only carbohydrate component of the membrane that is decreased during erythrocyte ageing, the others membranous sugars are decreased too. Theses changes in the carbohydrate moity could have a role in the clearance of the erythrocytes. 相似文献
9.
An improved, fast, and relatively simple procedure for isolation of hepatic mitochondrial contact sites is described. These contact sites include conventional outer membrane, but the inner membrane component (which we term fusion patches) has a unique biochemical composition characterized by a clustering of three specific inner membrane proteins of 54, 52, and 31 kDa identified by proteomics, respectively, as the alpha and beta subunits of ATP synthase and the liver isoform of adenine nucleotide transferase. The contact site fraction was prepared using a discontinuous sucrose gradient from crude outer membranes derived from swollen/shrunk rat liver mitochondria. The resultant contact sites were analyzed using a continuous sucrose density gradient, revealing an apparent heterogeneity due to varying amounts of retained fusion patches in relation to the unvarying outer membrane component. By electron microscopy, contact sites consist of small vacuoles that contain one or several tiny vesicles, many of which are composed of multiple, closely packed lamellae. The contact site subfraction morphology is consistent with the biochemical variation. Thus, contact sites are not haphazard fusions of outer and inner membrane, but consist in part of regions of inner membrane of novel composition (fusion patches) and of conventional outer membrane. 相似文献
10.
Michael Trauner Marco Arrese Martin Wagner 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2010,1801(3):299-310
Fatty liver disease comprises a spectrum ranging from simple steatosis to steatohepatitis which can progress to liver cirrhosis and hepatocellular cancer. Hepatic lipotoxicity may ensue when the hepatic capacity to utilize, store and export fatty acids (FA) as triglycerides is overwhelmed. Additional mechanisms of hepatic lipotoxicity include abnormal FA oxidation with formation of reactive oxygen species, disturbances in cellular membrane FA and phospholipid composition, alterations of cholesterol content and ceramide signalling. Lipotoxicity is a key factor for the progression of fatty liver disease by inducing hepatocellular death, activating Kupffer cells and an inflammatory response, impairing hepatic insulin signalling resulting in insulin resistance, and activation of a fibrogenic response in hepatic stellate cells that can ultimately lead to cirrhosis. Therefore, the concept of hepatic lipotoxicity should be considered in future therapeutic concepts for fatty liver disease. 相似文献
11.
J Tsagurnis H Kolb K Heininger J Schlepper-Sch?fer 《Biochemical and biophysical research communications》1985,130(3):1002-1009
We have tested whether mannose- and galactose-specific lectins on liver cells are able to bind antibody-antigen complexes and thus function as Fc-receptors. Rat hepatocytes and liver sinusoidal cells were isolated by collagenase perfusion and differential centrifugation. Rat erythrocytes were coated with purified IgM or IgG from rabbits immunized with rat erythrocytes. Both IgM and IgG coated erythrocytes bound to liver macrophages but not to hepatocytes. The binding of IgM and IgG coated red blood cells to liver macrophages could not be blocked by potent inhibitors for mannose- and galactose-specific macrophage lectins such as mannan, D-mannose-bovine serum albumin, N-acetyl-D-galactosamine, D-galactose-bovine serum albumin, or asialofetuin. Although lectin activity is calcium dependent and trypsin sensitive neither condition blocked rosette formation between liver macrophages and opsonized erythrocytes. Thus mannose- and galactose-specific lectins are not involved in the sequestration of IgM- or IgG-antibody-erythrocyte complexes in the liver. 相似文献
12.
In the course of a freeze-cleave study on intercellular junctions in the regenerating rat liver, we observed an unusual array of intramembranous particles located in regions of contact between endothelial cells lining the hepatic sinusoids. These arrays were characterized by an accumulation of particles which resembled a zonula occludens in their linear deployment but differed in that the contact regions were composed of individual particles which remained separated from each other by regular particle-free intervals. 相似文献
13.
Hubert Kolb Victoria Kolb-Bachofen 《Biochemical and biophysical research communications》1978,85(2):678-683
Rat Kupffer cells in vitro strongly bind neuraminidase-treated rat erythrocytes but not untreated erythrocytes. Binding between cells is inhibited by preincubation of macrophages with D-galactose and related sugars, but not with unrelated saccharides. We therefore suggest that cell adherence is mediated by a galactose-specific receptor on the Kupffer cell membrane. 相似文献
14.
Enhanced maintenance and functions of rat hepatocytes induced by combination of on-site oxygenation and coculture with fibroblasts 总被引:4,自引:0,他引:4
Nishikawa M Kojima N Komori K Yamamoto T Fujii T Sakai Y 《Journal of biotechnology》2008,133(2):253-260
In in vivo liver tissue, each hepatocyte has intimate interactions not only with adjacent hepatocytes but also with nonparenchymal cells in a three-dimensional (3D) manner. We recently reported that hepatic function is highly maintained on collagen covalently immobilized poly-dimethylsiloxane (PDMS) membranes through which oxygen is supplied directly to the cells. In this study, to further enhance performances of hepatocytes culture, we investigated cocultivation of rat hepatocytes with a mouse fibroblast, NIH/3T3 (3T3) in the same PDMS membranes. Various functions of hepatocytes were better maintained on the membrane at remarkably higher levels, particularly albumin secretion on such coculture was about 20 times higher than that in conventional coculture on tissue-culture-treated polystyrene (TCPS) surfaces. The remarkable functional enhancements are likely to be explained by the net growth of hepatocytes (from 1.2- to 1.4-fold inoculated number) and very intimate contact between hepatocytes and 3T3 cells in almost continuous double-layered structures under the adequate oxygen supply. The results demonstrate that simultaneous realization of different requirements toward mimicking in vivo liver tissue microstructure is effective in improving performance of hepatocytes culture system. 相似文献
15.
H M Hughes G M Powell D J Snodin J W Daniel A Crawford J K Sanders C G Curtis 《The Biochemical journal》1989,259(2):537-543
The metabolism of 14C- and 36Cl-labelled 1,6-dichloro-1,6-dideoxyfructose (DCF) was studied in the isolated perfused rat liver system. Dechlorination of DCF occurred in the liver and erythrocytes and was GSH-dependent. The GSH conjugate formed was identified by 13C and 1H n.m.r. as the 6-chlorofructos-1-yl-SG conjugate. It is proposed that the GS- anion attacks the low steady-state concentration of the reactive keto form of DCF and that the conjugate formed cyclizes to the dominant beta-anomer. 6-Chlorofructos-1-yl-SG conjugate of hepatic origin is excreted into bile, whereas that produced in erythrocytes does not enter the liver. 相似文献
16.
大鼠运动性疲劳模型的建立 总被引:5,自引:0,他引:5
目的建立大鼠运动疲劳模型,观察运动疲劳对大鼠各项生理、生化指标的影响。方法20只大鼠随机分为正常对照组和运动疲劳模型组,运动疲劳模型组大鼠每日按照方案进行锻炼。实验结束后大鼠检测相关指标:血清MDA含量和红细胞SOD活性,肝脏与骨骼肌MDA含量、SOD活性,骨骼肌线粒体游离钙离子含量,骨骼肌线粒体膜电位,下丘脑神经递质。电镜观察骨骼肌线粒体细微结构。结果运动疲劳模型组大鼠造模2周以后其血清、肝和骨骼肌MDA含量均有显著升高,红细胞和骨骼肌SOD活性均有显著降低,骨骼肌线粒体膜电位显著性降低,骨骼肌线粒体游离Ca2+含量有显著性降低,下丘脑GABA、5-HT含量有显著升高,下丘脑DA、ACh含量有显著性下降,电镜观察显示骨骼肌超微结构改变并以线粒体改变较为明显。结论大鼠跑台运动2周可造成运动疲劳模型,并造成大鼠骨骼肌线粒体损伤。 相似文献
17.
Epimorphin is involved in differentiation of rat hepatic stem-like cells through cell-cell contact 总被引:5,自引:0,他引:5
Miura K Nagai H Ueno Y Goto T Mikami K Nakane K Yoneyama K Watanabe D Terada K Sugiyama T Imai K Senoo H Watanabe S 《Biochemical and biophysical research communications》2003,311(2):415-423
Epimorphin, a mesenchymal cell surface-associated molecule, is detected on hepatic stellate cells (HSCs) in the liver. Here, we show the involvement of epimorphin in differentiation of rat hepatic stem-like cells (HSLCs) through contact with HSCs. HSLCs, isolated from adult rats, cultured in stellate cell-conditioned medium had no phenotypic and morphological changes, whereas HSLCs co-cultured with HSCs expressed albumin, transferrin, and tyrosine aminotransferase. An anti-epimorphin antibody inhibited hepatocytic differentiation of HSLCs in co-culture. Furthermore, epimorphin induced mRNA expression of albumin, transferrin, tyrosine aminotransferase, and gamma-glutamyl transpeptidase with decrease of c-kit and musashi-1. Morphologically, HSLCs piled up when co-cultured with HSCs, which was dramatically inhibited by an anti-epimorphin antibody. HSLCs contact with epimorphin started piling up, changed their shape from flat to cuboidal, and subsequently developed bile-canaliculi-like structures. In conclusion, epimorphin is a factor that induces differentiation of hepatic stem-like cells through epithelial-mesenchymal cell contact. 相似文献
18.
We used an immunohistochemical method to examine the localization of monoamine oxidase type B (MAOB) in rat liver. At the light microscopic level, MAOB was highly expressed in rat liver. It was intense around portal area, and weak around central area. All the hepatocytes examined had MAOB immunoreactivity. For the first time, using a double-labeling immunofluorescence histochemical method for laser microscopy, we report that no MAOB is found in endothelial cells, hepatic stellate cells, or Kupffer's cells. When examined under transmission electron microscopy, MAOB was localized to the mitochondrial outer membrane of hepatocytes. No apparent localization of MAOB was found in the rough endoplasmic reticulum, the crystal membrane of mitochondria, the nuclear envelope, or the plasma membrane. 相似文献
19.
E Friedrich H Sinn R Kühnlein R Carubelli 《Virchows Archiv. B, Cell pathology including molecular pathology》1978,29(3):201-209
Xenogeneic and enzyme-treated erythrocytes rapidly home to the rat liver as measured by scintigraphic procedures. Histology and scanning electron microscopy revealed that pronase digested erythrocytes are taken up as whole cells by liver histiocytes. 相似文献
20.
Sara Vander Borght Louis Libbrecht Aezam Katoonizadeh Jos van Pelt David Cassiman Frederik Nevens Alfons Van Lommel Bryon E Petersen Johan Fevery Peter L Jansen Tania A Roskams 《The journal of histochemistry and cytochemistry》2006,54(9):1051-1059
Breast cancer resistance protein (BCRP/ABCG2) is an ATP-binding cassette transport protein that is expressed in several organs including the liver. Previous studies have shown that ABC transport proteins play an important pathophysiological role in several liver diseases. However, to date, expression pattern and possible role of BCRP in human liver diseases and animal models have not been studied in detail. Here we investigated the expression pattern of BCRP in normal liver, chronic parenchymal and biliary human liver diseases, and parallel in different rat models of liver diseases. Expression was studied by immunohistochemistry and additionally by RT-PCR analysis in Thy-1-positive rat oval cells. Bile ducts, hepatic progenitor cells, reactive bile ductules, and blood vessel endothelium were immunoreactive for BCRP in normal liver and all types of human liver diseases and in rat models. BCRP was expressed by the canalicular membrane of hepatocytes in normal and diseased human liver, but never in rat liver. Remarkably, there was also expression of BCRP at the basolateral pole of human hepatocytes, and this was most pronounced in chronic biliary diseases. In conclusion, BCRP positivity in the progenitor cells/reactive ductules could contribute to the resistance of these cells to cytotoxic agents and xenotoxins. Basolateral hepatocytic expression in chronic biliary diseases may be an adaptive mechanism to pump bile constituents back into the sinusoidal blood. Strong differences between human and rat liver must be taken into account in future studies with animal models. 相似文献