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R Montesano 《The American journal of anatomy》1975,144(3):387-391
During a freeze-fracture study of tight junctions in fetal rat liver (Montesano et al., '75) unusual patterns of intramembranous particles were observed in regions of contact between sinusoidal endothelial cells. These patterns were mainly represented by arrays of particles, often associated with linear elevations or crests in the membrane A-face; they may represent abortive forms of tight junctions. 相似文献
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Soumya Ivaturi Catherine J. Wooten Maikhanh D. Nguyen Gene C. Ness Dayami Lopez 《Biochemical and biophysical research communications》2014
Several findings suggest that the low-density lipoprotein (LDL) receptor may internalize different lipoprotein particles via diverse pathways. Using a combination of discontinuous sucrose gradients and Triton solubilization studies, we demonstrated that the LDL receptor could be located simultaneously in clathrin-coated pits and caveolae in rat and human liver and in human hepatocyte-like C3A cells. Treatment with the cholesterol biosynthesis inhibitor, zaragozic acid A, shifted the distribution of the LDL receptor to clathrin containing fractions, whereas treatment with cholesterol or LDL shifted the receptor distribution towards caveolin-1 containing fractions. The LDL-dependent shift of the LDL receptor to caveolae coincided with a reduction in internalization of Bodipy-LDL. Redistribution within plasma membrane microdomains in response to specific treatments resulting in changes in LDL receptor function represents a novel paradigm that could be exploited in the development of a new class of therapeutic drugs. 相似文献
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Uptake of phosphatidylserine-containing liposomes by liver sinusoidal endothelial cells in the serum-free perfused rat liver 总被引:2,自引:0,他引:2
We studied the kinetics of hepatic uptake of liposomes during serum-free recirculating perfusion of rat livers. Liposomes consisted of phosphatidylcholine, cholesterol and phosphatidylserine in a 6:4:0 or a 3:4:3 molar ratio and were radiolabelled with [3H]cholesteryl oleyl ether. The negatively charged liposomes were taken up to a 10-fold higher extent than the neutral ones. Hepatic uptake of fluorescently labelled liposomes was examined by fluorescence microscopy. The neutral liposomes displayed a typical Kupffer cell distribution pattern, in addition to weak diffuse staining of the parenchyma, while the negatively charged liposomes showed a characteristic sinusoidal lining pattern, consistent with an endothelial localization. In addition, scattered Kupffer cell staining was distinguished as well as diffuse parenchymal fluorescence. The mainly endothelial localisation of the negatively charged liposomes was confirmed by determining radioactivity in endothelial and Kupffer cells isolated following a 1-h perfusion. Perfusion in the presence of polyinosinic acid, an inhibitor of scavenger receptor activity, reduced the rate of uptake of the negatively charged liposomes twofold, indicating the involvement of this receptor in the elimination mechanism. These results are compatible with earlier in vitro studies on liposome uptake by isolated endothelial cells and Kupffer cells, which showed that in the absence of serum also endothelial cells in situ are able to take up massive amounts of negatively charged liposomes. The present results emphasize that the high in vitro endothelial cell uptake in the absence of serum from earlier observations was not an artifact induced by the cell isolation procedure. 相似文献
5.
We studied the kinetics of hepatic uptake of liposomes during serum-free recirculating perfusion of rat livers. Liposomes consisted of phosphatidylcholine, cholesterol and phosphatidylserine in a 6:4:0 or a 3:4:3 molar ratio and were radiolabelled with [3H]cholesteryl oleyl ether. The negatively charged liposomes were taken up to a 10-fold higher extent than the neutral ones. Hepatic uptake of fluorescently labelled liposomes was examined by fluorescence microscopy. The neutral liposomes displayed a typical Kupffer cell distribution pattern, in addition to weak diffuse staining of the parenchyma, while the negatively charged liposomes showed a characteristic sinusoidal lining pattern, consistent with an endothelial localization. In addition, scattered Kupffer cell staining was distinguished as well as diffuse parenchymal fluorescence. The mainly endothelial localisation of the negatively charged liposomes was confirmed by determining radioactivity in endothelial and Kupffer cells isolated following a 1-h perfusion. Perfusion in the presence of polyinosinic acid, an inhibitor of scavenger receptor activity, reduced the rate of uptake of the negatively charged liposomes twofold, indicating the involvement of this receptor in the elimination mechanism. These results are compatible with earlier in vitro studies on liposome uptake by isolated endothelial cells and Kupffer cells, which showed that in the absence of serum also endothelial cells in situ are able to take up massive amounts of negatively charged liposomes. The present results emphasize that the high in vitro endothelial cell uptake in the absence of serum from earlier observations was not an artifact induced by the cell isolation procedure. 相似文献
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《Cell biology international reports》1989,13(5):419-425
Divergent viewpoints have been expressed regarding the existence of free coated vesicles in animal cells; this question has not been carefully addressed with plant tissues. Soybean suspension culture protoplasts were exposed to cationized ferritin (CF) for short times to label coated pits and coated vesicles. A serial section analysis did not reveal deep coated pits with long necks as reported in animal cells. Serial sections clearly demonstrated CF-labelled coated vesicles to be separate organelles and is consistent with the idea that they transfer CF from coated pits to other cytoplasmic organelles during endocytosis. 相似文献
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H J Schlayer M Woort-Menker S Eyhorn H Becker H E Schaefer K Decker 《Virchows Archiv. B, Cell pathology including molecular pathology》1988,55(4):225-232
Beta-glucuronidase and N-AS-D-chloroacetate esterase cytochemistry have been applied to rat liver sinusoidal endothelial cells and Kupffer cells. Both staining procedures allowed a clear-cut differentiation of either cell type. Kupffer cells which had been stained with beta-glucuronidase showed a positive reaction, whereas sinusoidal endothelial cells were completely negative. If the chloroacetate reaction was used, the former stained diffusely while the latter showed a characteristic granular staining pattern. Identity and purity of sinusoidal endothelial cells and Kupffer cells was validated by transmission and scanning electron microscopy as well as by the pattern of released eicosanoids which is characteristic for either cell type. These two staining techniques are a valuable addition to the peroxidase reaction commonly applied for differentiation. 相似文献
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H. -J. Schlayer M. Woort-Menker S. Eyhorn H. Becker H. E. Schaefer K. Decker 《Virchows Archiv. B, Cell pathology including molecular pathology》1988,55(1):225-232
Beta-glucuronidase and N-AS-D-chloroacetate esterase cytochemistry have been applied to rat liver sinusoidal endothelial cells
and Kupffer cells. Both staining procedures allowed a clear-cut differentiation of either cell type. Kupffer cells which had
been stained with beta-glucuronidase showed a positive reaction, whereas sinusoidal endothelial cells were completely negative.
If the chloroacetate reaction was used, the former stained diffusely while the latter showed a characteristic granular staining
pattern. Identity and purity of sinusoidal endothelial cells and Kupffer cells was validated by transmission and scanning
electron microscopy as well as by the pattern of released eicosanoids which is characteristic for either cell type. These
two staining techniques are a valuable addition to the peroxidase reaction commonly applied for differentiation. 相似文献
9.
Falkowska-Hansen B Oynebråten I Uhlin-Hansen L Smedsrød B 《Molecular and cellular biochemistry》2006,287(1-2):43-52
We have previously reported that liver sinusoidal endothelial cells (LSECs) are responsible for the clearance of monocyte chondroitin sulfate proteoglycan serglycin from the circulation (øynebråten et al.(2000) J. Leukocyte Biol. 67; 183–188). The aim of the present study was to investigate the kinetics of degradation of endocytosed serglycin in primary cultures of LSECs. The final degradation products of serglycin labelled biosynthetically in the glycosaminoglycan (GAG) chains with [3H] in the acetyl groups of N-acetyl galactosamine residues, [14C] in the pyranose rings, or [35S] in the sulfate groups were identified as[3H]-acetate, [14C]-lactate and [35S]-sulfate. Comparison of the rate of release of degradation products from the cells after endocytosis of serglycin labelled chemically with 125I in the tyrosine residues, or biosynthetically with [35S] or [3H] in the sulfate or acetyl groups, respectively, showed that 125I appeared more rapidly in the medium than [35S]-sulfate and [3H]-acetate. Judging from the speed of appearance of free 125I both intracellularly and in the medium, the core protein is degraded considerably more rapidly than the GAG chains.Desulfation of the GAG chains starts after the GAG chains are released from the core protein. Generation of lactate and acetate as the final products from degradation of the carbon skeleton of the GAG chains indicates that catabolism of endocytosed macromolecules in LSECs proceeds anaerobically. 相似文献
10.
Summary In the present study we have localized neutral phosphatase, acid phosphatase, alkaline phosphatase and 5 nucleotidase in the sinusoidal cells of rat liver using enzyme cytochemistry at light and electron microscopical level.Neutral phosphatase was present in the endoplasmic reticulum and nuclear envelope of parenchymal cells and of sinusoidal endothelial, Kupffer and fat-storing cells. The intensity of the neutral phosphatase reaction was stronger in sinusoidal than in parenchymal cells. Sinusoidal cells were devoid of cytochemically demonstrable alkaline phosphatase. Abundant acid phosphatase was present in the many lysosomes of endothelial and Kupffer cells. Substanually less acid phosphatase-positive lysosomes were found in fat-storing cells. 5 nucleotidase was present on the cell membrane of fat-storing cells, on 90% of all Kupffer cells and on the microvilli of parenchymal cells.We have further shown that combined staining for 5 nucleotidase and for endogenous peroxidase, offers a histochemical tool to discriminate between the three main sinusoidal cell types in normal rat liver. 相似文献
11.
Extremely rapid endocytosis mediated by the mannose receptor of sinusoidal endothelial rat liver cells. 总被引:3,自引:1,他引:2
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Isolated sinusoidal endothelial rat liver cells (EC) in suspension bound and internalized ovalbumin, a mannose-terminated glycoprotein, in a saturable manner. The binding and uptake were Ca2+-dependent and were effectively inhibited by alpha-methyl mannoside and yeast mannan, but not by galactose or asialoglycoproteins. This corresponds to the binding specificity described for the mannose receptor of macrophages and non-parenchymal liver cells. Binding studies indicated a surface pool of 20,000-25,000 mannose receptors per cell, with a dissociation constant of 6 x 10(-8) M. Uptake and degradation of ovalbumin by isolated EC were inhibited by weak bases and ionophores which inhibit acidification of endocytic vesicles and dissociation of receptor-ligand complexes. Cycloheximide had no effect on uptake or degradation. Degradation, but not uptake, was inhibited by leupeptin. We conclude that ovalbumin dissociates from the mannose receptors in the endosomal compartment and the receptors are recycled to the cell surface, while the ovalbumin is directed to the lysosomes for degradation. A fraction of the internalized ovalbumin was recycled intact to the cell surface and escaped degradation (retroendocytosis). The rate of internalization of ovalbumin by isolated EC was very fast, with a Ke (endocytotic rate constant) of 4.12 min-1, which corresponds to a half-life of 10 s for the surface pool of receptor-ligand complexes. To our knowledge, this is the highest Ke reported for a receptor-mediated endocytosis system. 相似文献
12.
肝窦内皮细胞(liver sinusoidal endothelial cells,LSECs)是肝脏抵御炎症和免疫反应的第一道防线,其独特的窗孔结构及功能特性决定它在肝纤维化发生发展中起重要作用。LSECs主要通过介导肝脏炎症反应、肝窦毛细血管化、活化肝星状细胞(hepatic stellate cells,HSC)、引发细胞外基质(extracellular matrix,ECM)的生成与降解失衡等途径参与肝纤维化的发生发展。靶向LSECs对治疗肝纤维化极具潜力,阐述清楚二者调控关系,将为抗肝纤维化治疗提供新的理论依据。 相似文献
13.
The process of clathrin-mediated endocytosis tightly regulates signaling of the superfamily of seven-transmembrane G protein-coupled receptors (GPCRs). A fundamental question in the cell biology of membrane receptor endocytosis is whether activated receptors can initiate the formation of clathrin-coated pits (CPs) or whether they are simply mobilized to pre-existing CPs. Here, using various approaches, including a dynamic assay to monitor the distribution of CPs and GPCR-beta-arrestin complexes in live HeLa cells, we demonstrate for the first time that activated GPCRs do not initiate the de novo formation of CPs but instead are targeted to pre-existing CPs. 相似文献
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Culture and characterization of sinusoidal endothelial cells isolated from human liver 总被引:2,自引:0,他引:2
George W. Daneker Serena A. Lund S. Wright Caughman Robert A. Swerlick Andrew H. Fischer Charles A. Staley Edwin W. Ades 《In vitro cellular & developmental biology. Animal》1998,34(5):370-377
Summary Although most vascular models use large vessel endothelial cells from human umbilical veins, there is marked heterogeneity
among endothelial cells from different vascular beds and organs. More accurate modeling of endothelial involvement in liver
diseases, including metastasis, may result from the use of human hepatic sinusoidal endothelial cells. Liver resection specimens
were sectioned, then treated with a 1.2 U/ml dispase solution. The tissue slurry was mechanically disaggregated and separated
by centrifugation on a Percoll density gradient. Cells were then cultured in an endothelial-specific media with growth factors.
These techniques resulted in a homogeneous monolayer consistent with endothelial cells by light microscopy. An endothelial
origin was further confirmed by the expression of Factor VIII, binding of Ulex lectin, and uptake of acetylated low density lipoprotein. Electron microscopy showed transcellular fenestrations consistent
with a sinusoidal origin. These human hepatic sinusoidal endothelial cells were then studied for expression of the adhesion
molecules CD31/PECAM, CD34, E-selectin, ICAM-1, L-selectin, LFA-3, P-selectin, and VCAM-1 plus the binding of wheat germ agglutinin
lectin. The patterns of adhesion molecule expression and lectin binding by these cells are characteristic of hepatic sinusoidal
endothelia. In this paper, we have described a method for isolation and culture of human cells with the morphologic and phenotypic
characteristics of hepatic sinusoidal endothelia. 相似文献
17.
Zhaoli Tan Keyan Chen Yong Shao Lihua Gao Yan Wang Jianming Xu Yang Jin Xianwen Hu Youliang Wang 《Development, growth & differentiation》2016,58(7):620-631
Although liver sinusoidal endothelial cells (LSECs) have long been known to contribute to liver regeneration following injury, the exact role of these cells in liver regeneration remains poorly understood. In this work, we performed lineage tracing of LSECs in mice carrying Tie2‐Cre or VE‐cadherin‐Cre constructs to facilitate fate‐mapping of LSECs in liver regeneration. Some YFP‐positive LSECs were observed to convert into hepatocytes following a two‐thirds partial hepatectomy (PH). Furthermore, human umbilical vein endothelial cells (HUVECs) could be triggered to convert into cells that closely resembled hepatocytes when cultured with serum from mice that underwent an extended PH. These findings suggest that mature non‐hepatocyte LSECs play an essential role in mammalian liver regeneration by converting to hepatocytes. The conversion of LSECs to hepatocyte‐like (iHep) cells may provide a new approach to tissue engineering. 相似文献
18.
The endocytosis of latex particles (0.33, 0.46 and 0.80 micron in diameter) in the sinusoidal endothelial and Kupffer cells of the rat liver was studied electron microscopically. When the liver was perfused with serum-free oxygenated Krebs Ringer bicarbonate, latex particles of all three sizes were taken up by the endothelial cells. After a 10-min perfusion, particles were incorporated by the luminal cell surface of the perikarya or of the thick portion of the endothelial cells. A large patch of bristle coat was surrounding the ingested particle. The number of ingested particles in the endothelial cells, however, was much less than in the Kupffer cells. In in vivo experiments, no endocytosis of the latex particles was observed in the endothelial cells. In the Kupffer cells, particles were engulfed by the ruffled membranes or sank into the cytoplasm without a large patch of the bristle coat both in the perfusion system and in vivo. These observations show that at least 0.80 micron latex particles are taken up by the bristle-coated membranes in the sinusoidal endothelial cells of the perfused liver. The endocytic mechanism for latex particles in the endothelial cells is different from that of the Kupffer cells. 相似文献
19.
Yuan Peng Qian Chen Tao Yang Yanyan Tao Xiong Lu Chenghai Liu 《Molecular biology reports》2014,41(3):1815-1827
Cultured mycelium Cordyceps sinensis (CMCS) was widely used for a variety of diseases including liver injury, the current study aims to investigate the protective effects of CMCS on liver sinusoidal endothelial cells (LSECs) in acute injury liver and related action mechanisms. The mice were injected intraperitoneally with lipopolysaccharide (LPS) and d-galactosamine (D-GalN). 39 male BABL/c mice were randomly divided into four groups: normal control, model control, CMCS treatment and 1,10-phenanthroline treatment groups. The Serum liver function parameters including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were assayed with the commercial kit. The inflammation and scaffold structure in liver were stained with hematoxylin and eosin and silver staining respectively. The LSECs and sub-endothelial basement membrane were observed with the scanning and transmission electronic microscope. The protein expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in liver were analyzed with Western blotting. Expression of von Willebrand factor (vWF) was investigated with immunofluorescence staining. The lipid peroxidation indicators including antisuperoxideanion (ASAFR), hydroxyl free radical (·OH), superoxide dismutase (SOD), malondialdehyde and glutathione S-transferase (GST) were determined with kits, and matrix metalloproteinase-2 and 9 (MMP-2/9) activities in liver were analyzed with gelatin zymography and in situ fluorescent zymography respectively. The model mice had much higher serum levels of ALT and AST than the normal mice. Compared to that in the normal control, more severe liver inflammation and hepatocyte apoptosis, worse hepatic lipid peroxidation demonstrated by the increased ASAFR, ·OH and MDA, but decreased SOD and GST, increased MMP-2/9 activities and VCAM-1, ICAM-1 and vWF expressions, which revealed obvious LSEC injury and scaffold structure broken, were shown in the model control. Compared with the model group, CMCS and 1,10-phenanthroline significantly improved serum ALT/AST, attenuated hepatic inflammation and improved peroxidative injury in liver, decreased MMP-2/9 activities in liver tissue, improved integration of scaffold structure, and decreased protein expression of VCAM-1 and ICAM-1. CMCS could protect LSECs from injury and maintain the microvasculature integration in acute injured liver of mice induced by LPS/D-GalN. Its action mechanism was associated with the down-regulation of MMP-2/9 activities and inhibition of peroxidation in injured liver. 相似文献
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Hang TC Lauffenburger DA Griffith LG Stolz DB 《American journal of physiology. Gastrointestinal and liver physiology》2012,302(3):G375-G388
Primary rat liver sinusoidal endothelial cells (LSEC) are difficult to maintain in a differentiated state in culture for scientific studies or technological applications. Relatively little is known about molecular regulatory processes that affect LSEC differentiation because of this inability to maintain cellular viability and proper phenotypic characteristics for extended times in vitro, given that LSEC typically undergo death and detachment around 48-72 h even when treated with VEGF. We demonstrate that particular lipid supplements added to serum-free, VEGF-containing medium increase primary rat liver LSEC viability and maintain differentiation. Addition of a defined lipid combination, or even oleic acid (OA) alone, promotes LSEC survival beyond 72 h and proliferation to confluency. Moreover, assessment of LSEC cultures for endocytic function, CD32b surface expression, and exhibition of fenestrae showed that these differentiation characteristics were maintained when lipids were included in the medium. With respect to the underlying regulatory pathways, we found lipid supplement-enhanced phosphatidylinositol 3-kinase and MAPK signaling to be critical for ensuring LSEC function in a temporally dependent manner. Inhibition of Akt activity before 72 h prevents growth of SEC, whereas MEK inhibition past 72 h prevents survival and proliferation. Our findings indicate that OA and lipids modulate Akt/PKB signaling early in culture to mediate survival, followed by a switch to a dependence on ERK signaling pathways to maintain viability and induce proliferation after 72 h. We conclude that free fatty acids can support maintenance of liver LSEC cultures in vitro; key regulatory pathways involved include early Akt signaling followed by ERK signaling. 相似文献