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Immunology and Infection

Standardized Colon Ascendens Stent Peritonitis in Rats - a Simple, Feasible Animal Model to Induce Septic Acute Kidney Injury

Published: February 15th, 2022

DOI:

10.3791/54448

1Department of General, Visceral, Transplantation, Vascular and Paediatric Surgery, Department of Surgery I, University of Würzburg, 2Department of Anesthesiology and Critical Care, Medical Center-University of Freiburg, 3Faculty of Medicine, University of Freiburg, 4Department of Internal Medicine I, Division of Nephrology, University of Würzburg, 5Department of Physiology and Pharmacology, West Virginia University School of Medicine, Robert C. Byrd Health Science Center, 6Department of Anesthesiology and Intensive Care Medicine, Robert-Bosch-Krankenhaus

Acute kidney injury (AKI) is a severe complication in critically ill patients and is related with an increased mortality. Here, we present a reliable and reproducible in vivo model to mimic AKI under inflammatory conditions that might contribute towards understanding the pathogenesis of septic AKI.

AKI in septic patients is associated with increased mortality and poor outcome despite major efforts to refine the understanding of its pathophysiology. Here, an in vivo model is presented that combines a standardized septic focus to induce AKI and an intensive care (ICU) setup to provide an advanced hemodynamic monitoring and therapy comparable in human sepsis. Sepsis is induced by standardized colon ascendens stent peritonitis (sCASP). AKI is investigated functionally by measurement of blood and urine samples as well as histologically by evaluation of histopathological scores. Furthermore, the advanced hemodynamic monitoring and the possibility of repetitive blood gas sampling enable a differentiated analysis of severity of induced sepsis.

The sCASP method is a standardized, reliable and reproducible method to induce septic AKI. The intensive care setup, continuous hemodynamic and gas exchange monitoring, low mortality rate as well as the opportunity of detailed analyses of kidney function and impairments are advantages of this setup. Therefore, the described method may serve as a new standard for experimental investigations of septic AKI.

Sepsis still remains the leading cause of death on non-cardiac intensive care units (ICU) with mortality rates of ≈ 30 - 50%1,2,3. A hallmark of severe sepsis and septic shock is the acute kidney injury that causes a further increase of mortality rate when it is associated with distant organ dysfunction such as cardiac and respiratory failure4,5,6. The overall incidence of AKI in ICU patients varies from 20 to 50%7. Despite the pivotal role of AKI regard....

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All animal procedures were approved by the Laboratory Animal Care and Use Committee of the District of Unterfranken, Germany and carried out according to the Declaration of Helsinki.

1. Surgical preparation and installation of invasive monitoring and continuous medication

  1. Anesthetize Sprague-Dawley rats using isoflurane delivered by precision vaporizer at a concentration and flow rate approved by local institution’s animal research oversight body and/or veterinary team. Confirm adequate depth of .......

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As previously published by Schick et al.8, we demonstrate the following results.

Induction of sepsis without mortality
In the CASP model, sepsis is induced by a continuous leakage of intraluminal located bacteria of the colon ascendens into the abdominal cavity resulting in fecal peritonitis and bacteremia. Hereby, the size of the implanted catheter regulates the output of faeces an.......

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The pathophysiology of septic AKI still remains unknown in its complexity. Clinical research and trials in patients will not enable gains of new insights with respect to histopathology changes, microcirculation disturbances or drug interactions on cellular levels15. It has been postulated previously that there is a need for improved and new animal models to investigate acute kidney injury associated with sepsis19. Therefore, we established a new animal model for septic AKI .......

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M.A. Schick and N. Schlegel received funding from the Deutsche Forschungsgemeinschaft (DFG) SCHL 1962/2-1 and SCHL 1962/4-1.

....

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Name Company Catalog Number Comments
Sprague-Dawley rats Janvier Labs, France
 Isoflurane CP cp-pharma, Burgdorf, Germany
polyethylen catheter PE 10; 30m A. Hartenstein, Wuerzburg, Germany 0.58x0.96 mm
Swivel (375/D/20) Instech, Plymouth Meeting, PA, USA (375/D/20)
plastic button tethers Instech, Plymouth Meeting, PA, USA LW105S
Perfusor B. Braun; Melsungen, Germany Perfusor fm
suction catheter ch. 10 B.Braun Melsungen AG, Germany suction catheter typy „Ideal“; ch. 10
suture Syneture; USA Surgipro; Monofilament Polypropylen 4-0
suture Ethicon; Scotland Prolene; Polypropylen 5-0
14G-i.v. catheter BD Insynte; BD Vialon; Madrid; Spain 14GA i.v. catheter
cotton buds NOBA Verbandmittel Danz GmbH u Co KG; Wetter; Germany
rodent respirator Hugo Sachs Elektronik KG, Germany rodent respirator, Type:7025
Midazolam Ratiopharm, Germany Midazolam
Thermodilutioncatheter ADInstruments, Spechbach, Germany
p-Aminohippuric acid Sigma-Aldrich; St. Louis; USA p-Aminohippuric acid sodium salt; A3759-25G
Inulin Sigma-Aldrich; St. Louis; USA Inulin-FITC; F3272-1G
Formaldehyde Otto Fischar GmbH & CoKG; Saarbrücken, Germany Formaldehyde 3.5%
Cyclopentan Merck; Darmstadt; Germany Uvasol: 2-Methylbutan
alcohol based scrub Schülke & Mayr GmbH, Norderstedt; Germany kodan Tinktur forte; 45g 2-Propanol, 10g 1-Propanol per 100g
povidone iodine solution B.Braun Melsungen AG, Germany Braunol, 7.5g Povidone Iodine per 100g

  1. Angus, D. C., et al. Epidemiology of severe sepsis in the United States: analysis of incidence, outcome, and associated costs of care. Crit Care Med. 29, 1303-1310 (2001).
  2. Dellinger, R. P., et al. Surviving sepsis campaign: international guidelines for management of severe sepsis and septic shock: 2012. Crit Care Med. 41, 580-637 (2013).
  3. Moerer, O., Quintel, M. [Sepsis in adult patients - definitions, epidemiology and economic aspects]. Internist (Berl). 50, 788-796 (2009).
  4. Belayev, L. Y., Palevsky, P. M. The link between acute kidney injury and chronic kidney disease. Curr Opin Nephrol Hypertens. 23, 149-154 (2014).
  5. Chao, C. T., et al. The impact of dialysis-requiring acute kidney injury on long-term prognosis of patients requiring prolonged mechanical ventilation: nationwide population-based study. PLoS One. 7, e50675 (2012).
  6. Chertow, G. M., Christiansen, C. L., Cleary, P. D., Munro, C., Lazarus, J. M. Prognostic stratification in critically ill patients with acute renal failure requiring dialysis. Arch Intern Med. 155, 1505-1511 (1995).
  7. Doi, K. Role of kidney injury in sepsis. J Intensive Care. 4, 17 (2016).
  8. Gomez, H., et al. A unified theory of sepsis-induced acute kidney injury: inflammation, microcirculatory dysfunction, bioenergetics, and the tubular cell adaptation to injury. Shock. 41, 3-11 (2014).
  9. Deitch, E. A. Animal models of sepsis and shock: a review and lessons learned. Shock. 9, 1-11 (1998).
  10. Esmon, C. T. Why do animal models (sometimes) fail to mimic human sepsis?. Crit Care Med. 32, S219-S222 (2004).
  11. Rittirsch, D., Hoesel, L. M., Ward, P. A. The disconnect between animal models of sepsis and human sepsis. J Leukoc Biol. 81, 137-143 (2007).
  12. Doi, K., Leelahavanichkul, A., Yuen, P. S., Star, R. A. Animal models of sepsis and sepsis-induced kidney injury. J Clin Invest. 119, 2868-2878 (2009).
  13. Zantl, N., et al. Essential role of gamma interferon in survival of colon ascendens stent peritonitis, a novel murine model of abdominal sepsis. Infect Immun. 66, 2300-2309 (1998).
  14. Maier, S., et al. Cecal ligation and puncture versus colon ascendens stent peritonitis: two distinct animal models for polymicrobial sepsis. Shock. 21, 505-511 (2004).
  15. Schick, M. A., et al. Sepsis-induced acute kidney injury by standardized colon ascendens stent peritonitis in rats - a simple, reproducible animal model. Intensive Care Med Exp. 2, (2014).
  16. Schick, M. A., et al. Effects of crystalloids and colloids on liver and intestine microcirculation and function in cecal ligation and puncture induced septic rodents. BMC Gastroenterol. 12, 179 (2012).
  17. Schneider, R., et al. Downregulation of organic anion transporters OAT1 and OAT3 correlates with impaired secretion of para-aminohippurate after ischemic acute renal failure in rats. Am J Physiol Renal Physiol. 292, F1599-F1605 (2007).
  18. Schick, M. A., et al. The impact of crystalloid and colloid infusion on the kidney in rodent sepsis. Intensive Care Med. 36, 541-548 (2010).
  19. Bellomo, R., Ronco, C., Kellum, J. A., Mehta, R. L., Palevsky, P. Acute renal failure - definition, outcome measures, animal models, fluid therapy and information technology needs the Second International Consensus Conference of the Acute Dialysis Quality Initiative (ADQI) Group. Crit Care. 8, R204-R212 (2004).
  20. Lustig, M. K., et al. Colon ascendens stent peritonitis--a model of sepsis adopted to the rat: physiological, microcirculatory and laboratory changes. Shock. 28, 59-64 (2007).
  21. Traeger, T., et al. Colon ascendens stent peritonitis (CASP)--a standardized model for polymicrobial abdominal sepsis. J Vis Exp : JoVE. , (2010).
  22. Dyson, A., Rudiger, A., Singer, M. Temporal changes in tissue cardiorespiratory function during faecal peritonitis. Intensive Care Med. 37, 1192-1200 (2011).
  23. Arakelyan, K., et al. Early effects of an x-ray contrast medium on renal T(2) */T(2) MRI as compared to short-term hyperoxia, hypoxia and aortic occlusion in rats. Acta physiol. 208 (2), 202-213 (2013).
  24. Schick, M. A., et al. Balanced hydroxyethylstarch (HES 130/0.4) impairs kidney function in-vivo without Inflammation. PLoS One. 10 (9), e0137247 (2015).
  25. Flemming, S., et al. Phosphodiesterase 4 inhibition dose dependently stabilizes microvascular barrier functions and microcirculation in a rodent model of polymicrobial sepsis. Shock. 41, 537-545 (2014).
  26. Flemming, S., et al. Sphingosine-1-phosphate receptor-1 agonist sew2871 causes severe cardiac side effects and does not improve microvascular barrier breakdown in sepsis. Shock. 49 (1), 71-81 (2018).

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