scholarly journals A new method to monitor Kupffer-cell function continuously in the perfused rat liver. Dissociation of glycogenolysis from particle phagocytosis

1990 ◽  
Vol 266 (1) ◽  
pp. 141-147 ◽  
Author(s):  
K B Cowper ◽  
R T Currin ◽  
T L Dawson ◽  
K A Lindert ◽  
J J Lemasters ◽  
...  
Keyword(s):  
Rat Liver ◽  
Kupffer Cell ◽  
Kupffer Cells ◽  
Cell Function ◽  
New Method ◽  
Carbon Uptake ◽  
Prostaglandin D2 ◽  
Perfused Rat Liver ◽  
Colloidal Carbon ◽  
Parenchymal Cells

In order to study particle phagocytosis and glycogenolysis simultaneously, this study was designed to develop a direct-read-out method to monitor Kupffer-cell function continuously, based on the uptake of colloidal carbon by the isolated perfused rat liver. Livers were perfused for 20 min with Krebs-Henseleit buffer saturated with O2/CO2 (19:1). Colloidal carbon (1-2 mg/ml) was added to the buffer, and absorbance of carbon was monitored continuously at 623 nm in the effluent perfusate. Since colloidal-carbon uptake was proportional to A623, rates of uptake were determined from the influent minus effluent concentration difference, the flow rate and the liver wet weight. Rates of colloidal-carbon uptake were 50-200 mg/h per g and were proportional to the concentration of carbon infused. Data from light-microscopy and cell-separation studies demonstrated that carbon was taken up exclusively by non-parenchymal cells and predominantly by Kupffer cells. Further, the amount of colloidal carbon detected histologically in non-parenchymal cells increased as the concentration of colloidal carbon in the perfusate was elevated. When Kupffer cells were activated or inhibited by treatment with endotoxin or methyl palmitate, carbon uptake was increased or decreased respectively. Taken together, these results indicate that Kupffer-cell function can be monitored continuously in a living organ. This new method was utilized to compare the time course of phagocytosis of carbon by Kupffer cells and carbohydrate output by parenchymal cells. Carbohydrate output increased rapidly by 69 +/- 9 mumol per g within 2-4 min after addition of carbon and returned to basal values within 12-16 min. However, carbon uptake by the liver did not reach maximal rates until about 15 min. Infusion of a cyclo-oxygenase inhibitor, aspirin (10 mM), caused a progressive decrease in carbohydrate output and blocked the stimulation by carbon completely. Aspirin neither altered rates of carbon uptake nor prevented stimulation of carbohydrate release by addition of N2-saturated buffer. The data from these experiments are consistent with the hypothesis that output of mediators by Kupffer cells, presumably prostaglandin D2 and E2, occurs transiently as Kupffer cells begin to phagocytose foreign particles in the intact organ, a process which continues at high rates for hours.

Dose-dependent effects of acute lindane treatment on Kupffer cell function assessed in the isolated perfused rat liver

Xenobiotica ◽  
1997 ◽  
Vol 27 (7) ◽  
pp. 747-757 ◽  
Author(s):  
L. A. VIDELA* ◽  
P. TRONCOSO ◽  
A. C. M. ARISI ◽  
V. B. C. JUNQUEIRA
Keyword(s):  
Rat Liver ◽  
Kupffer Cell ◽  
Cell Function ◽  
Isolated Perfused Rat Liver ◽  
Perfused Rat Liver ◽  
Dose Dependent

Stimulation of oxygen uptake by prostaglandin E2 is oxygen dependent in perfused rat liver

1998 ◽  
Vol 275 (3) ◽  
pp. G542-G549 ◽  
Author(s):  
Wei Qu ◽  
Zhi Zhong ◽  
Gavin E. Arteel ◽  
Ronald G. Thurman
Keyword(s):  
Oxygen Uptake ◽  
Oxygen Tension ◽  
Rat Liver ◽  
Kupffer Cells ◽  
High Flow ◽  
Perfused Rat Liver ◽  
Normal Flow ◽  
Flow Rates ◽  
Glucose Output ◽  
Parenchymal Cells

The aim of this study was to determine if the effect of prostaglandin E2(PGE2) on hepatic oxygen uptake was affected by oxygen tension. Livers from fed female Sprague-Dawley rats were perfused at normal or high flow rates (4 or 8 ml ⋅ g−1 ⋅ min−1) to vary local oxygen tension within the liver lobule. During perfusion at normal flow rates, PGE2 (5 μM) infusion increased oxygen uptake by about 50 μmol ⋅ g−1 ⋅ h−1; however, when livers were perfused at high flow rates, the increase was nearly twice as large. Simultaneously, glucose output was increased rapidly by about 50%, whereas glycolysis was decreased about 60%. When flow rate was held constant, increases in oxygen uptake due to PGE2 were proportional to oxygen delivery. Infusion of PGE2 into livers perfused at normal flow rates increased state 3 rates of oxygen uptake of subsequently isolated mitochondria by about 25%; however, rates were increased 50–75% in mitochondria isolated from livers perfused at high flow rates. Thus it is concluded that PGE2stimulates oxygen uptake via mechanisms regulated by oxygen tension in perfused rat liver. High flow rates also increased basal rates of oxygen uptake: this increase was prevented by inactivation of Kupffer cells with GdCl3. In addition, conditioned medium from Kupffer cells incubated at high oxygen tension (75% oxygen) stimulated oxygen uptake of isolated parenchymal cells by >30% and elevated PGE2production about twofold compared with Kupffer cells exposed to normal air-saturated buffer (21% oxygen). These effects were blocked completely by both indomethacin and nisoldipine. These data support the hypothesis that oxygen stimulates Kupffer cells to release mediators such as PGE2 which elevate oxygen consumption in parenchymal cells, possibly by mechanisms involving cyclooxygenase and calcium channels.

A new method to monitor Kupffer cell phagocytosis continuously in perfused rat liver

Hepatology ◽  
1991 ◽  
Vol 13 (3) ◽  
pp. 567-574 ◽  
Author(s):  
Patricia E. Ganey ◽  
Barbara Keller ◽  
Steven N. Lichtman ◽  
John J. Lemasters ◽  
Ronald G. Thurman
Keyword(s):  
Rat Liver ◽  
Kupffer Cell ◽  
New Method ◽  
Perfused Rat Liver

scholarly journals Hepatic lipase is localized at the parenchymal cell microvilli in rat liver

1997 ◽  
Vol 321 (2) ◽  
pp. 425-430 ◽  
Author(s):  
Belinda BREEDVELD ◽  
Kees SCHOONDERWOERD ◽  
Adrie J. M. VERHOEVEN ◽  
Rob WILLEMSEN ◽  
Hans JANSEN
Keyword(s):  
Endothelial Cells ◽  
Rat Liver ◽  
Kupffer Cells ◽  
Binding Capacity ◽  
Hepatic Lipase ◽  
Parenchymal Cell ◽  
Liver Cells ◽  
Minor Amount ◽  
Parenchymal Liver ◽  
Parenchymal Cells

Hepatic lipase (HL) is thought to be located at the vascular endothelium in the liver. However, it has also been implicated in the binding and internalization of chylomicron remnants in the parenchymal cells. In view of this apparent discrepancy between localization and function, we re-investigated the localization of HL in rat liver using biochemical and immunohistochemical techniques. The binding of HL to endothelial cells was studied in primary cultures of rat liver endothelial cells. Endothelial cells bound HL in a saturable manner with high affinity. However, the binding capacity accounted for at most 1% of the total HL activity present in the whole liver. These results contrasted with earlier studies, in which non-parenchymal cell (NPC) preparations had been found to bind HL with a high capacity. To study HL binding to the different components of the NPC preparations, we separated endothelial cells, Kupffer cells and blebs by counterflow elutriation. Kupffer cells and endothelial cells showed a relatively low HL-binding capacity. In contrast, the blebs, representing parenchymal-cell-derived material, had a high HL-binding capacity (33 m-units/mg of protein) and accounted for more than 80% of the total HL binding in the NPC preparation. In contrast with endothelial and Kupffer cells, the HL-binding capacity of parenchymal cells could account for almost all the HL activity found in the whole liver. These data strongly suggest that HL binding occurs at parenchymal liver cells. To confirm this conclusion in situ, we studied HL localization by immunocytochemical techniques. Using immunofluorescence, we confirmed the sinusoidal localization of HL. Immunoelectron microscopy demonstrated that virtually all HL was located at the microvilli of parenchymal liver cells, with a minor amount at the endothelium. We conclude that, in rat liver, HL is localized at the microvilli of parenchymal cells.

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