Lipid and lipid-free total parenteral nutrition: differential effects on macrophage phagocytosis in rats

doi:10.1016/S0899-9007(99)00186-0

Nutrition

Volume 15, Issues 11–12, 1 November 1999, Pages 885-889

https://doi.org/10.1016/S0899-9007(99)00186-0 Get rights and content

Abstract

Lipid emulsions provided with total parenteral nutrition (TPN) have been associated with mononuclear phagocytic system functional changes. The aim of the present investigation was to assess the influence of TPN with added lipid emulsions on macrophage (Mφ) phagocytosis. Wistar rats (n = 70) with external jugular vein cannulation were randomized into seven groups. The rats received an oral diet or six different isocaloric (1.16 kcal/mL), isonitrogenous (1.5 g/mL), and isolipidic (30% non-protein calories) TPN regimens: (a) an oral diet with intravenous infusion of saline (OS); (b) non-lipid TPN (glucose); (c) TPN with 10% long chain triacylglycerol emulsion (LCT); (d) TPN with 90% LCT and 10% fish oil (FO) emulsion; (e) TPN with 50% LCT and 50% FO; (f) TPN with 10% lipid emulsion with 50% medium chain triacylglycerol (MCT) and 50% LCT; and (g) TPN with 45% MCT, 45% LCT, and 10% FO. After 96 h of TPN or saline infusion, colloidal carbon (Pelikan, Germany) was injected intravenously at 1.0 mL/kg body weight, and the rats were killed after 3 h. Liver, spleen, and lung were weighed and prepared by immunohistochemistry analyses with the HAM-56 anti-Mφ antibody. Under light microscopy, the total Mφ number (MT) and the colloidal carbon phagocytic Mφ number (MP) were established, and the phagocytic index was calculated as MP/MT × 100. There were no statistical (P < 0.05) differences in liver, spleen, or lung weights among the seven groups in comparison with the OS group. Non-lipid TPN inhibited spleen and lung Mφ phagocytosis when compared with the OS and lipid-TPN groups. Lipid TPN supplemented with fish oil emulsion increased total liver and lung Mφ number and phagocytosis. These results indicate that TPN supplemented with fish oil increases Mφ phagocytosis in rats.

Introduction

Total parenteral nutrition (TPN) is one of the great advances in clinical practice and is widely applied in hospital settings.1 In general, TPN consists of amino acids, glucose, and electrolytes. With parenteral nutrition, lipid emulsions (LE) are usually administered twice a week or daily to provide adequate essential fatty acids and calorie intake.

LE administration, however, can alter cell structure and activity. Phospholipid cell membranes are composed of phosphate groups and lipid components. The degree of saturation and fatty acid deficiency in phospholipid cell membranes may modify cell membrane structure and function by changing fluidity and regulation of protein mobility and favoring interaction of several substances with their specific receptors.

Important effects on the immune system, especially on the mononuclear phagocytic system (MPS), have been described with parenteral LE.2, 3 The use of methyl palmitate and ethyl esterate have inhibitory effects on the reticuloendothelial system and reduce resistance to lethal endotoxin doses in rats.4 Intravenous (IV) administration of fatty acid esters with 14–20 carbon atoms can also induce depression of MPS phagocytic activity.5

The physicochemical nature of the infused triacylglycerol (i.e., carbon chain length, saturation degree, size of particle, and nature of fatty acid) may determine structural changes and alterations in macrophage activity. Experimental lipid-TPN infusion for 7 d has been associated with decreased phagocytosis and increased superoxide production by peritoneal macrophages, splenic macrophage proliferation, and bacterial translocation.6

Intravenous infusion of LE containing long chain triacylglycerols (LCT) and a physical mixture of LCT and medium chain triacylglycerols (MCT) in rats infected by Escherichia coli (E. coli) intraperitoneal infusion was followed by hyperplasia of Kupffer cells and splenic histiocytes with both LE.7 The infusion of TPN containing LCT fat emulsion in guinea pigs reduced E. coli phagocytosis by Kupffer cells and splenic macrophages.3

Inhibition of pulmonary bacterial phagocytosis was observed in guinea pigs after infusion of TPN with LCT but not with TPN containing 75% non-protein calories as MCT/LCT LE. It was suggested that the liver reticuloendothelial system, mainly lung macrophages, was able to phagocytose circulating bacteria because it was not lipid inhibited as occurred with LCT LE.8 These changes were not reported when TPN containing 50% non-protein calories in the form of LCT was infused.9 In clinical practice, the inhibiting effect of LE on the functional activity of MPS, although still controversial, may have an important harmful impact because parenteral nutrition is increasingly recommended in hypercatabolic situations in which inflammatory and infectious processes may be present.

A new LE containing fish oil rich in ω-3 fatty acids has become available. The effects of this LE on macrophage (Mφ) function are not well known and merit investigation. ω-3 long chain fatty acids are less immunosupressive than ω-6 long chain fatty acids and they may maintain or improve Mφ functions.10

The goal of this study was to evaluate the influence of glucose and several lipid TPN regimens on Mφ phagocytosis in different organs of the MPS in rats.

Experimental studies regarding lipid parenteral emulsions on Mφ phagocytosis are controversial. These trials analyzed rats, guinea pigs, mice, liver Mφ, lung Mφ, peritoneal Mφ, LCT, or MCT/LCT emulsions in isolation.

Our proposal was to design an experimental study that could compare the effect of old and new fat emulsions in the framework of TPN to determine the effect on Mφ phagocytosis simultaneously in liver, spleen, and lung.

Section snippets

Materials and methods

Seventy adult male Wistar Norwegian albino rats (180.0–250.0 g) from the central vivarium of the University of São Paulo Medical School (FMUSP) were used. The animals were kept at room temperature under day-night light cycles in metabolic cages in the laboratory of Experimental Surgery Laboratory of FMUSP for 3 d of acclimation. All animals had free access to water and chow.

The animals were randomly separated into seven groups composed of 10 rats each. Central vein cannulation was performed in

Body weight and infusion volume

There were no statistical differences in the infused TPN volume across the seven experimental groups (Table IV). The animals fed an oral diet gained weight (P < 0.05), whereas animals fed the glucose or lipid TPN diets did not have a significant weight gain (Table IV).

Macrophage phagocytosis

The Mφ phagocytosis results are presented in Table V. Total liver Mφ number decreased with glucose (P < 0.05) and fat emulsion containing LCT (P < 0.05) as opposed to the other TPN infusion groups.

Spleen total Mφ number and

Discussion

This experimental study was performed to observe the colloidal carbon phagocytosis of macrophages in the liver, spleen, and lung after challenge with TPN containing isocaloric and isolipidic amounts of different LE. With the use of immunohistochemical methods to count the Mφ total number and phagocytosing Mφ number and establish a phagocytic index, it was possible to better understand the behavior of in situ Mφ than by indirect methods using radionuclides and isotopes.17, 18

In the current

Summary

The aim of the present investigation was to assess the influence of TPN with LE on Mφ phagocytosis. After 96 h of lipid and non-lipid TPN or saline infusion, liver, spleen, and lung were prepared by immunohistochemic analyses. Non-lipid TPN inhibited spleen and lung Mφ phagocytosis when compared with the oral-fed group and lipid-TPN groups. Lipid TPN supplemented with fish oil emulsion increased total liver and lung Mφ number and phagocytosis.

References (27)

M.M. Berger et al.
A 10-year survey of nutritional support in a surgical ICU1986–1995
Nutrition
(1997)
N. Di Luzio
Employment of lipids in the measurements and modifications of cellular, humoral and immune responses
Adv Lipid Res
(1972)
J. Shou et al.
Total parenteral nutrition, bacterial translocation, and host immune function
Am J Surg
(1994)
J. Sobrado et al.
Lipid emulsion and reticuloendothelial system function in healthy and burned guinea pigs
Am J Clin Nutr
(1985)
J. Tovar
Á
, Mahour GH, Miller SW, Isaacs H, Smith CN. Endotoxin clearance after intralipid infusion. J Pediatr Surg
(1976)
J.D. Palombo et al.
Rapid modulation of lung and liver macrophage phospholipid fatty acids in endotoxemic rats by continuous enteral feeding with n-3 and gamma-linoleic fatty acids
Am J Clin Nutr
(1996)
R.L. Jones et al.
Hematologic alterations in diabetes mellitus
Am J Med
(1981)
Z. Friedman et al.
Effect of parenteral fat emulsions on the pulmonary and reticuloendothelial systems in the newborn infant
Pediatrics
(1978)
H.J. Hamawy et al.
The effect of lipid emulsions on reticuloendothelial system function in the injured animal
JPEN
(1985)
A.E. Stuart et al.
Susceptibility of mice to bacterial endotoxin after modification of reticuloendothelial function by simple lipids
J Pathol Bacteriol
(1962)

D.L. Waitzberg et al.

Efeito de emulsões lipı́dicas sobre mecanismos de agressão infecciosa

Rev Hosp Clin Fac Med S Paulo

(1992)

M. Vallgren et al.

Effect of intralipid on clearance and organ uptake of bacteria in intra-abdominal sepsis rats

Clin Nutr

(1986)

J.D. Palombo et al.

Fatty acid composition of lung, macrophage and surfactant phospholipids after short-term enteral feeding with n-3 lipids

Lipids

(1994)

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This result demonstrated that TPLE-1 possessed lower toxicity compared to Taxol®. Excessive fat infusion could cause lung, spleen, liver reticuloendothelial cell deposition, to greatly weaken the normal phagocytosis and damage the immune system for tumor patients to need long-term treatment (Cukier et al., 1999; Seidner et al., 1989). In order to prepare TPLEs with different particle size, the oil content in our previous prescription of BLE was adjusted from 20% (w/v) to 10% (w/w).
The discovery of new intravenous drug delivery carrier for water-insoluble drug is a challenging task. In this paper, novel two-vial formulation of paclitaxel (PTX)-loaded lipid nanoemulsions (TPLEs) with particle sizes of 110 nm (TPLE-1), 220 nm (TPLE-2) and 380 nm (TPLE-3), which were formed by mixing a PEG400 solution of PTX and 10% (w/w) blank lipid emulsions (BLEs) with different particle size prior to use, were developed and comparatively evaluated for their pharmaceutics, pharmacokinetics, biodistribution, in vitro and in vivo anticancer efficiency. Among them, TPLE-1 displayed higher PTX-loading, slower PTX-release and larger PTX-distribution in oil-phase, significantly reduced extraction by RES organs, increased tumor-uptake, showed stronger cytotoxicity against MCF-7 cells and more potent anticancer efficacy on MCF-7 tumor-bearing nude mice, and had greater plasma AUC_0-∞ value, smaller plasma clearance (CL), longer mean residence time (MRT) and elimination half-life (T_1/2) in SD rats. It also exhibited the same in vivo efficacy as Taxol^® and even produced less hemolysis and intravenous irritation. Moreover, its LD₅₀ was 4.3-fold higher than that of Taxol^®. All results demonstrate that TPLE-1 is a promising candidate drug due to its high tumor-accumulation and effectiveness, low toxicity, good safety and druggability in clinical application for the cancer therapy.
Reduction of blood stream infections in children following a change to chlorhexidine disinfection of parenteral nutrition catheter connectors
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Excess lipid administration can result in hypertriglyceridemia and impairment of leukocyte function.26,27 Lipid metabolites may accumulate in hepatic sinusoidal macrophages over time, thereby interfering with clearance and enhancing toxicity of bacteria and bacterial products in the bloodstream.28 Secondly, there were a higher proportion of immune-compromised patients after stem cell or solid organ transplantation and thirdly children studied were younger and would have had less mature immune systems.
Catheter-related-blood-stream-infection (CRBSI) might be prevented by optimal catheter connector antisepsis in children with intestinal failure on parenteral nutrition (PN). We changed the disinfectant used from isopropanol 70% to chlorhexidine 2% in 70% isopropanol, which leaves a residue of chlorhexidine on the connector.
We conducted this before/after study in children treated with PN for >28 days. Episodes of CRBSI were recorded for all 42 children treated for >28 days during May–November 2006 before introducing chlorhexidine and for all 50 children treated in May–November 2007 after chlorhexidine was introduced.
The number of hospital-acquired CRBSI and number of PN days was counted for each period. The rate of CRBSI/1000 catheter days and the proportion of patients that experienced at least one CRBSI during the two periods were compared.
There were 3.1 CRBSI/1000 catheter days prior to using chlorhexidine and 0.4 CRBSI/1000 catheter days after it was introduced, p = 0.03.
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Our results support the use of 2% chlorhexidine not only to reduce risk of sepsis for central venous catheter connector antisepsis in catheters used for intravenous nutrition, but also to improve the patients' chances of recovering intestinal function.
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Thus, the plasma TG reduction, in soy/fish oil diet, may be caused primarily by reduced intestinal fat absorption and other unknown mechanisms, instead of VLDL synthesis and faster blood clearance. Although LPL plays important roles in TG-rich particle clearance, other pathways (macrophage phagocytosis, proteoglycans, etc.) may also be involved.33,34 In soy/fish oil diet, it was possibly that only increased LPL activity contributed to emulsion clearance, and this was not enough to enhance clearance rates.
Intake of n-3 fatty acids can reduce both fasting and postprandial triglyceride (TG) concentrations in humans as well as in experimental animals, but the mechanisms by which this occurs are not completely known. We investigated in mice the effects of dietary fish oil (a source of n-3 fatty acids) on endogenous TG synthesis and exogenous TG-rich particle removal.
C57 BL/6J mice were fed for 4 months with three types of high-fat diets (18% fat wt/wt) – soy oil, fish oil and a mixture of soy oil and fish oil (soy/fish) (5:1 wt/wt), and a chow diet with 6% fat from soy oil (wt/wt) served as a control. Plasma TG and apolipoprotein B (apoB) concentrations and lipoprotein lipase (LPL) activity were measured. Triton WR 1339 was used to assess hepatic synthesis of very low density lipoprotein, and intravenous injection of chylomicron-like lipid emulsions was conducted to determine the effects of dietary fish oil n-3 fatty acids on exogenous TG clearance.
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Our data suggest that reduced endogenous TG synthesis, increased LPL activities and more rapid blood clearance of TG-rich particles all distinctly contribute to the TG-lowering effects of fish oil n-3 fatty acids.
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Based on the results of the present study, we can suggest that the presence of FO does not constitute a factor influencing neutrophils migration in rats, associated to MCT/LCT or in SMOF LE composition. With relation to phagocytosis, the increased number of phagocytosing macrophages in the liver and lung of rats under LE infusion containing MCT/LCT and FO is in accordance with results reported in previous study of our laboratory with Wistar rats.9 Nevertheless, in the present study we have found an increased macrophages number in the spleen and increased phagocytosing macrophages in the liver and lung by MCT/LCT without FO supplementation, in contrast to our previous observations.9
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Influence of parenteral fat emulsion Intralipos<sup>®</sup> and citric acid on blood viscosity and erythrocyte morphology in vitro
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In most studies fat emulsion had been administered by enteral route. Recently a parenteral soybean-oil emulsion has been developed. In this study we assessed the effects of a parenteral soybean-oil emulsion (Intralipos^®) and citric acid on blood rheology and erythrocyte morphology in vitro. Porcine blood was incubated in vitro with increasing concentrations of fat emulsion Intralipos^® and citric acid at 37 °C for 1 h. Viscosity of plasma and whole blood was measured using a FASCO-2050 digital viscometer. Red blood cell morphology was examined by light microscopy. The viscosity of whole blood represented an ascending dose-dependence for different Intralipos^® concentrations at high shear rate of 90 and 225 s⁻¹, however, it decreased with citric acid concentrations. On the other hand, the whole blood viscosity also declined with citric acid concentrations in presence of 30% Intralipos^® (v/v), and there is a minimal viscosity at 0.67% citric acid (v/v). There are some thorns on the blood membrane at 40% Intralipos^® as compared with control (no Intralipos^® addition), which indicates the Intralipos^® compound may affect blood cell membranes, and resulted in whole blood viscosity increase. We concluded that the intravenous soybean-oil preparation Intralipos^® interacts with the erythrocyte membrane, and citric acid could alleviate the whole blood viscosity.
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^☆: This study was supported by grants 94/1614-0 and 95/1179-4 from FAPESP.

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Basic Nutritional InvestigationsLipid and lipid-free total parenteral nutrition: differential effects on macrophage phagocytosis in rats☆

Abstract

Introduction

Section snippets

Materials and methods

Body weight and infusion volume

Macrophage phagocytosis

Discussion

Summary

Nutrition

Adv Lipid Res

Am J Surg

Am J Clin Nutr

, Mahour GH, Miller SW, Isaacs H, Smith CN. Endotoxin clearance after intralipid infusion. J Pediatr Surg

Am J Clin Nutr

Am J Med

Effect of parenteral fat emulsions on the pulmonary and reticuloendothelial systems in the newborn infant

Pediatrics

The effect of lipid emulsions on reticuloendothelial system function in the injured animal

JPEN

Susceptibility of mice to bacterial endotoxin after modification of reticuloendothelial function by simple lipids

J Pathol Bacteriol