The impact of arginine on bacterial translocation in an intestinal obstruction model in rats

doi:10.1016/j.clnu.2006.12.007

Clinical Nutrition

Volume 26, Issue 3, June 2007, Pages 335-340

https://doi.org/10.1016/j.clnu.2006.12.007 Get rights and content

Summary

Background & aims

Arginine has been shown to have multiple beneficial metabolic and immunologic effects in stress situations. Supplementation of arginine has been shown to promote wound healing and intestinal mucosal recovery after trauma, ischemia or intestinal resection. Bacterial translocation has also been evaluated although with conflicting results and using different assessing techniques. Therefore, the aim of this study was to evaluate the effects of arginine on bacterial translocation in an intestinal obstruction model in rats using Escherichia coli labeled with ^99mTechnetium.

Methods

Male Wistar rats (250–350 g) were randomized to receive conventional chow, diet supplemented with pure arginine or diet supplemented with an immunonutrition enteral formula, enriched with arginine, omega-3 fatty acid and RNA. After 7 days, the animals were anesthetized. Terminal ileum was isolated and a ligature was placed around it. E. coli labeled with ^99mTechnetium (^99mTc-E. coli) was inoculated into the intestinal lumen (terminal ileum). After 24 h, the animals were sacrificed. Blood, mesenteric lymph nodes (MLN), liver, spleen and lungs were removed for radioactivity determination.

Results

Arginine supplementation (300 mg/day, 600 mg/day or present in the enteral formula) reduced the level of bacterial translocation when compared with the control group (p<0.05). This was shown by significantly decrease uptake of ^99mTc-E. coli in blood, MLN, liver, spleen and lungs of the animals in the experimental groups (p<0.05).

Conclusions

These results have shown that arginine was able to decrease bacteria translocation despite intestinal obstruction. There are several mechanisms which might explain the role of arginine and these will be the subject of future studies.

Section snippets

Background

Nutrition has usually been seen as the supply of energy and nitrogen to maintain body mass. Recent studies have, however, revealed that nutrition also plays an important role in modulating the immune and inflammatory responses. The clinical relevance of this modulation has led to the investigation of diets containing specific substrates the so-called immunonutrients, such as glutamine, arginine, nucleotides and omega-3 fatty acids.¹ These formulas have been related to modulation of gut

Animals

Forty male Wistar rats weighting between 250 and 350 g were included in this study. The animals were randomized to five groups of eight animals each one: (1) “sham” operated (no obstruction; inoculation of 12.5 MBq of ^99mTechnetium-Escherichia coli (^99mTc-E. coli)) fed a conventional chow diet (LABINA^®, Purina, Brazil; 23% protein); (2) IO group fed a conventional chow diet, submitted to intestinal obstruction and inoculation of 12.5 MBq of ^99mTc-E. coli; (3) Arginine 300 mg group fed conventional

Composition of standard chow and supplemented chows

The composition of standard and supplemented chows is shown in Table 1. There was an increase in total calories in the Impact formula (355.6 kcal/100 g) compared to the conventional chow (275 kcal/100 g) and the arginine supplemented chow (264.3 kcal/100 g).

There was an increase of protein, nitrogen and arginine in the chow supplemented with arginine and an increase of arginine, fat and total calories in the chow supplemented with the enteral formula. Carbohydrate content was slight reduced in all

Discussion

The role of arginine in preventing bacterial translocation in an experimental intestinal obstruction model was assessed. Although arginine has been reported to reduce bacterial translocation in experimental obstructive jaundice models by recovering viable organisms from the mesenteric lymph node (MLN) and spleen16, 17, 18 in our study we used ^99mTechnetium E. coli. This method is very fast, direct, simple and does not require aseptic conditions throughout the experiments.13, 15 Another key

Acknowledgments

Ajionomoto, Brazil and Novartis Brazil for having, respectively, given Arginine and the Impact formula. CNPQ (National Counsel of Technological and Scientific Development), Fundep (Fundação de Desenvolvimento da Pesquisa) for the grants.

Iara Eliza Pacífico Quirino contributed the conception and design of the experiment and experimental procedures, statistical analysis and interpretation of data; Maria Isabel T.D. Correia the conception and design of the study, interpretation of the data,

References (23)

G. O’Callaghan et al.
The role of immune-enhancing diets in the management of perioperative patients
Crit Care Resusc
(2003)
A. Gurbuz et al.
Supplemental dietary arginine accelerates intestinal mucosal regeneration and enhances bacterial clearance following radiation enteritis in rats
J Surg Res
(1998)
S. Ersin et al.
The prophylactic and therapeutic effects of glutamine and arginine enriched diets on radiation induced enteritis in rats
J Surg Res
(2000)
H.F. Shang et al.
Effects of arginine supplementation on mucosal immunity in rats with septic peritonits
Clin Nutr
(2004)
J. De Oca et al.
Selective bowel decontamination, nutritional therapy and bacterial translocation after burn injury
Clin Nutr
(1993)
R. Wiest et al.
Bacterial translocation in the gut
Best Pract Res Clin Gastroenterol
(2003)
B. Zulfikaroglu et al.
The effect of immunonutrition on bacterial translocation, and intestinal villus atrophy in experimental obstructive jaundice
Clin Nutr
(2003)
J. MacFie
Enteral versus parenteral nutrition: the significance of bacterial translocation and gut-barrier function
Nutrition
(2000)
J.Y. Wang et al.
Effect of arginine on gastrointestinal immunity during total parenteral nutrition
Clin Nutr
(1996)
I. Sukhotnik et al.
Oral arginine reduces gut mucosal injury caused by lipopolysaccharide in rat
J Surg Res
(2004)

G.P. Zaloga

Immune-enhancing enteral diets: where's the beef?

Crit Care Med

(1998)

Cited by (31)

L-ARGININE PREVENTS ISCHEMIC INJURY IN EXPLANTED RAT INTESTINAL REGIONS IN AN EX VIVO PERFUSION MODEL
2022, Transplantation Reports
Citation Excerpt :
Inhibition of NOS by L-NAME decreases the production of NO and reduces the protective effect of l-arginine against IRI [12]. L-arginine has been shown to play an important beneficial role in protection from the inflammatory response of the small intestine and it is involved in leukocyte adherence associated with IRI, decreasing the levels of free radicals, inhibition of neutrophil-endothelium interactions, prevention of bacterial translocation and, ultimately, helps in the preservation of intestinal barrier integrity [13,14,15,16]. Previous studies have focused on preconditioning of either the donor or the recipient of small bowel transplants as a means of diminishing the effect of IRI and promoting intestinal regional viability [10,17].
The small intestine is one of the most sensitive organs to ischemia. L-arginine has been shown to reduce damage from ischemia and reperfusion injury. We hypothesize that explanted intestinal segments from rats will demonstrate reduced susceptibility to ischemic injury when perfused with l-arginine.
45 small intestinal segments were harvested from male Sprague-Dawley rats and connected to an ex vivo intestinal perfusion device. Ischemic damage was induced by perfusing the extraluminal side with Ringer-HEPES buffer saturated with 100% N2. All segments were then perfused intraluminally with and without l-arginine. We conducted a set of experiments with intraluminal perfusion with both l-arginine and N-nitroarginine methyl ester (L-NAME), an inhibitor of the nitric oxide – arginine pathway. Control segments were perfused extraluminally under non-ischemic conditions and intraluminally with and without l-arginine. The intraluminal perfusate contained FITC-inulin, and the fluorescence signal of FITC-inulin was measured to calculate average fluid secretion, which directly corresponds to the extent of ischemic injury.
Intestinal segments perfused with l-arginine had significantly decreased secretion over time in comparison to intestinal segments perfused without l-arginine (p<0.0001). Perfusion with L-NAME abrogated the protective effect of l-arginine.
Intraluminal perfusion with l-arginine reduced ischemic damage to harvested intestine.
Identification of Metabolic Changes in Ileum, Jejunum, Skeletal Muscle, Liver, and Lung in a Continuous I.V. Pseudomonas aeruginosa Model of Sepsis Using Nontargeted Metabolomics Analysis
2019, American Journal of Pathology
Citation Excerpt :
The semiessential amino acid arginine is beneficial under stress conditions,50 promoting nitrogen retention and mucosal barrier integrity,51 along with the release of anabolic hormones and improved wound healing.52 Enteral nutrition supplementation of arginine53,54 with glutamine has been reported to be more effective in protecting the intestinal mucosa in cases of endotoxemia.55–58 The findings in the present study raise the possibility that the benefits of supplementing with linoleic acid and arginine may be due to metabolic defects seen in the small intestine (ileum and jejunum).
Sepsis is a multiorgan disease affecting the ileum and jejunum (small intestine), liver, skeletal muscle, and lung clinically. The specific metabolic changes in the ileum, jejunum, liver, skeletal muscle, and lung have not previously been investigated. Live Pseudomonas aeruginosa, isolated from a patient, was given via i.v. catheter to pigs to induce severe sepsis. Eighteen hours later, ileum, jejunum, medial gastrocnemius skeletal muscle, liver, and lung were analyzed by nontargeted metabolomics analysis using gas chromatography/mass spectrometry. The ileum and the liver demonstrated significant changes in metabolites involved in linoleic acid metabolism: the ileum and lung had significant changes in the metabolism of valine/leucine/isoleucine; the jejunum, skeletal muscle, and liver had significant changes in arginine/proline metabolism; and the skeletal muscle and lung had significant changes in aminoacyl-tRNA biosynthesis, as analyzed by pathway analysis. Pathway analysis also identified changes in metabolic pathways unique for different tissues, including changes in the citric acid cycle (jejunum), β-alanine metabolism (skeletal muscle), and purine metabolism (liver). These findings demonstrate both overlapping metabolic pathways affected in different tissues and those that are unique to others and provide insight into the metabolic changes in sepsis leading to organ dysfunction. This may allow therapeutic interventions that focus on multiple tissues or single tissues once the relationship of the altered metabolites/metabolism to the underlying pathogenesis of sepsis is determined.
Malnutrition, immunity and infection
2013, Diet, Immunity and Inflammation
It has been over 50 years since Scrimshaw, Taylor and Gordon set out the bases for interpretation of synergistic and antagonistic interactions between malnutrition and infection. Any contemporary update must recognize emerging diseases such as HIV and re-emerging diseases such as tuberculosis. Genetic and epigenetic concepts and techniques have merged with the advance of technology in basic and applied immunity. Not only nutrient deficiencies, but also their excesses – including obesity and overweight – alter host defenses and risk of infections. Issues of vitamin D, host disease and infection, and interaction of the microbiome of fecal flora with the gut immune system represent two of the exciting contemporary departures.
Influence of uracil on bacterial translocation in an intestinal obstruction model in rats
2013, International Journal of Surgery
Citation Excerpt :
This is clearly seen in 99mTc-E. coli uptake of U1 with U2. Furthermore, Quirino et al. suggested that arginine also reduced the bacterial translocation, but the amount of arginine had no effect on bacterial translocation.6 On the other hand, Batista and colleagues have investigated the effect of citrulline on bacterial translocation and found that citrulline decreased bacterial translocation, which matches our results.22
Bacterial translocation occurs when intestinal mucosa and the intestinal wall lose their barrier properties against bacteria such as in the case of intestinal obstruction. Enteral nutrition with immunonutrients strengthens the immune system and thickens the intestinal barrier thus preventing bacterial translocation.
The purpose of this study is to investigate the effect of uracil which is an immunonutrient on bacterial translocation using rats with intestinal obstruction as a model.
Wistar albino rats were divided into three groups. The control group was fed with standard chow diet, while the other two groups were fed with uracil-supplemented chow diet. The rats were fed with these diets for seven days. And the end of the feeding period all groups were submitted intestinal obstruction and injected with ^99mTc labeled Escherichia coli into the rats' terminal ileum under anesthetic. The rats were sacrificed 24 h later. Their blood, mesenteric lymph nodes (MLN), liver, spleen, lung and ileum were removed to determine level of radioactivity.
When compared with the control group it was determined that uracil supplementation reduced the level of bacterial translocation.
Uracil may be used in the prevention of bacterial translocation in cases of intestinal obstruction because it strengthens the intestinal barrier and the immune system. However, more studies are needed to clearly explain the mechanism behind uracil's beneficial role here.
Interactions between ω3 polyunsaturated fatty acids and arginine on nutritional and immunological aspects in severe inflammation
2010, Clinical Nutrition
Immune-enhancing diets (IEDs) contain a mixture of nutrients claimed to have immunological properties. Therefore, it seemed relevant to determine the effect of each of their components. The aim of this study was to examine the role of arginine (Arg) and ω3 polyunsaturated fatty acids (ω3 PUFAs) in the effect of an IED (Crucial^®) in a validated rat model of inflammation induced by turpentine (TI).
Forty-two rats were randomized into five groups: AL (ad libitum), TI-EN (TI + standard enteral nutrition (EN): Sondalis^®HP), TI-EN-Arg (TI + standard EN + Arg in equimolar concentration to Arg in the IED), TI-M-IED (TI + modified IED containing the same ω6/ω3 ratio as in standard EN) and TI-IED (TI + Crucial^®). Blood was sampled to determine CD25 receptor density on lymphocytes. TNF-α, IL-6 and NO (production and expression) were evaluated on isolated macrophages. Mesenteric lymph nodes, spleen and liver were cultured for analysis of enterobacterial translocation and dissemination.
CD25 density was decreased after TI and was corrected in the TI-EN-Arg, TI-M-IED and TI-IED groups (p < 0.05). TI induced an alteration of macrophage mRNA expression of IL-6, TNF-α and iNOS, corrected in the TI-EN-Arg and TI-M-IED groups (p < 0.05), but not by the IED. Enterobacterial translocation was observed in all treated groups, nevertheless the amount tended (p = 0.054) to be lower in the TI-EN-Arg group.
Arg and ω3 PUFAs make a major contribution to IED effects, but our study shows interaction between them on macrophage reactivity. This indicates that the individual properties of each pharmaconutrient are not additive in IEDs.
Pretreatment with arginine preserves intestinal barrier integrity and reduces bacterial translocation in mice
2010, Nutrition
Citation Excerpt :
BT has been studied in animals subjected to various pathological conditions such as hemorrhagic shock [22], endotoxin-challenge [23], thermal body surface trauma [24], acute pancreatitis [25], ischemia, and intestinal obstruction [26]. The latter, has been shown to cause disruption of intestinal epithelium, with subsequent increase of mucosal permeability and thus BT [10,11,21,26,27]. However, BT is a normal process that is essential for local and systemic immunity regulation [28,29].
To evaluate the effects of arginine on intestinal barrier integrity and bacterial translocation (BT) in mice undergoing intestinal obstruction.
Mice were divided into 3 groups, treated for 7 d before surgical intervention with isocaloric and isoprotein diets. The ARG group received a diet containing 2% arginine, the IO (intestinal obstruction) and Sham groups, standard chow diet. On the eighth day of treatment, all animals received diethylenetriamine pentaacetic acid (DTPA) solution labeled with ^99mTechnetium (^99mTc-DTPA) by gavage for intestinal permeability analysis. After 90 min, the animals were anesthetized and the terminal ileum ligated. The Sham group only underwent laparotomy. After 4, 8, and 18 h, blood was collected for radioactivity determination. Samples of ileum were collected 18 h after surgery for histological analysis. In another set of animals, BT was evaluated. After 7 d of treatment, all animals received 10⁸ CFU/mL of ^99mTc-E.coli by gavage; 90 min later they were submitted to the surgical procedure described above. BT was determined by the uptake of ^99mTc-E.coli in blood, mesenteric lymph nodes, liver, spleen, and lungs, assessed 18 h after the surgery.
The intestinal permeability and BT were higher in the IO group when compared with the Sham group (P < 0.05). Arginine supplementation reduced intestinal permeability and BT to physiologic levels. Histological analysis showed mucosal ileum preservation in animals treated with arginine.
Arginine was able to preserve barrier integrity, thus reducing BT.

View all citing articles on Scopus

View full text

Original ArticleThe impact of arginine on bacterial translocation in an intestinal obstruction model in rats

Summary

Background & aims

Methods

Results

Conclusions

Section snippets

Background

Animals

Composition of standard chow and supplemented chows

Discussion

Acknowledgments

Crit Care Resusc

J Surg Res

J Surg Res

Clin Nutr

Clin Nutr

Best Pract Res Clin Gastroenterol

Clin Nutr

Nutrition

Clin Nutr

J Surg Res

Immune-enhancing enteral diets: where's the beef?

Crit Care Med

Original Article
The impact of arginine on bacterial translocation in an intestinal obstruction model in rats