Acute Renal Failure (ARF) / Ischemia Reperfusion Injury (IR Injury)

Acute renal failure (ARF) is characterized by the sudden loss of the renal function. ARF is a frequent clinical problem, particularly in the intensive care medicine. Severe blood loss during major operations, sepsis, cardio-thoracic surgeries and radio-contrast media application are common causes for ARF. Underlying mechanisms for ARF include cell adhesion, cell infiltration with generation of oxygen free radicals (ROS), and inflammatory cytokine production.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In Vivo Models

To mimic acute renal failure IR injury can be induced in rodents by bilateral renal pedical clamping for 30-45 min, resulting in severe loss of renal function within 24h post-ischemia. Histologically ARF is characterized by tubular necrosis, tubular epithelial cell detachment, and tubular obstruction in the outer medullary stripe.

Available Readouts

  1. Survival

  2. Systemic and local inflammatory response

    • Cytokine / chemokine profile in serum and kidney tissue

  3. Renal function

    • clinical chemistry for serum creatinine, BUN

  4. Renal morphology

    • Tubular atrophy

    • Inflammation

    • Immunophenotyping of infiltrating leukocytes

  5. Immunohistochemistry and RT-PCR for specific markers of interests

  6. Apoptosis

  7. Renal damage marker KIM-1 and NGAL expression

 

 

 

 

 

 

 

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Renal Diseases

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Techniques

Chronic kidney disease

Progressive tubulo-interstitial fibrosis is the common end point leading to end-stage renal disease regardless of the etiology in experimental and clinical settings. Ureter obstruction due to nephrolithiasis, ureter occlusion by either enhanced scaring as radiation side effect or prostate hypertrophy in elder male patients or ureter valve insufficiency with urine congestion in young children with consecutive hydronephrosis can lead to progressive renal damage and chronic kidney disease (CKD). Also, acute kidney injury (AKI) in the context of major cardiac surgery is an important risk factor for progressive CKD. Currently, no effective treatment strategy to protect kidney function is available.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In Vivo Models

  • AKI induced CKD model

Unilateral renal pedical clamping to induce ischemia reperfusion injury (IRI) for 45 min causes progressive CKD within 2 - 3 weeks. This model mimics the clinical situation of AKI induced CKD. Follow up is 3 - 4 weeks.

 

  • Unilateral ureter obstruction (UUO) induced CKD model

Unilateral ureter obstruction (UUO) causes pressure induced progressive renal fibrosis. This model mimics the ureter obstruction due to nephrolithiasis, ureter occlusion by either enhanced scaring as radiation side effect or prostate hypertrophy in elder male patients or ureter valve insufficiency with urine congestion in young children with consecutive hydronephrosis. Follow up is 5 - 10 days.

  • 5/6 Nephrectomie induced CKD model

In this model the lower branch of the left renal artery is ligated to produce about one third area with visible renal ischemia; the upper pole of the left kidney is removed by cautery and the right kidney is decapsulated and nephrectomized to induce a total 5/6 nephrectomy. This leads to a progressive renal failure with severe glomerulosclerosis within 8-12 weeks. One important feature of renal injury in this particular model is hypertension, which alone perpetuates endothelial activation, inflammation, and proliferation, followed by vascular obliteration and glomerulosclerosis.

Available Readouts

  1. Renal morphology

    • Tubular atrophy

    • Inflammation

    • Immunophenotyping of infiltrating leukocytes

    • Extracellular matrix protein deposition

  2. Systemic and local inflammatory response

    • Cytokine / chemokine profile in serum and kidney tissue

  3. Fibrosis

    • Immunohistochemistry for fibronectin, a-SMA, collagen IV

    • Expression of profibrotic mediators

  4. Endothelial injury

    • Expression of adhesion molecule

  5. Renal damage marker KIM-1 and NGAL expression

  6. Immunohistochemistry for specific markers of interests

  7. Apoptosis

  8. Blood pressure​​​

Need further Readouts or cell lines? We routinely develop new models to fit our cutomer needs.

 

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Techniques

  1. Histology (H&E, Masson Trichrome, Periodic Acid Schiff (PAS) staining, Picro Sirius Red staining for collagen I/III)

  2. Immunohistochemistry

  3. Flow cytometry including Bead-based flow cytometry

  4. ELISA

  5. Molecular biology (Real-Time PCR, Western blot)

 

 

 

Renal failure - In Vitro Models

We use either cell lines or primary cell cultures.

 

 

 

 

 

 

 

Renal failure - relevant Publications

A novel therapy to attenuate acute kidney injury and ischemic allograft damage after allogenic kidney transplantation in mice.

Gueler F, Shushakova N, Mengel M, Hueper K, Chen R, Liu X, Park JK, Haller H, Wensvoort G, Rong S. PLoSOne. 2015 Jan 24; 10(1): e0115709. PMID: 25617900

Renal PKC-e deficiency attenuates acute kidney injury and ischemic allograft injury via TNF-a-dependent inhibition of apoptosis and inflammation.

Rong S, Hueper K, Kirsch T, Greite R, Klemann C, Mengel M, Meier M, Menne J, Leitges M, Susnik N, Meier M, Haller H, Shushakova N, Gueler F. Am J Physiol Renal Physiol. 2014 Sep 15; 307(6): F718 - 26. PMID: 25056349

C57BL/6 and 129/Sv mice: genetic difference to renal ischemia-reperfusion.

Lu X, Li N, Shushakova N, Schmitt R, Menne J, Susnik N, Meier M, Leitges M, Haller H, Gueler F, Rong S. J Nephrol. 2012 Sep - Oct; 25(5): 738 - 43. PMID: 22180224

Autoimmunity in CD73/Ecto-5'-nucleotidase deficient mice induces renal injury.

Blume C, Felix A, Shushakova N, Gueler F, Falk CS, Haller H, Schrader J. PLoS One. 2012; 7(5): e37100. PMID: 22666342

Bβ(15-42) attenuates the effect of ischemia-reperfusion injury in renal transplantation.

Sörensen I, Rong S, Susnik N, Gueler F, Shushakova N, Albrecht M, Dittrich AM, von Vietinghoff S, Becker JU, Melk A, Bohlmann A, Reingruber S, Petzelbauer P, Haller H, Schmitt R. J Am Soc Nephrol. 2011 Oct; 22(10): 1887 - 96. PMID: 21841063

Statins attenuate ischemia-reperfusion injury by inducing heme oxygenase-1 in infiltrating macrophages.

Gueler F, Park JK, Rong S, Kirsch T, Lindschau C, Zheng W, Elger M, Fiebeler A, Fliser D, Luft FC, Haller H. Am J Pathol. 2007 Apr; 170(4): 1192 - 9. PMID: 17392159

Peritoneal Dialysis

Peritoneal dialysis (PD) is a renal replacement therapy which uses the peritoneum as a membrane through which fluid and dissolved substances are exchanged with the blood. This is an important treatment option for a worldwide growing number of patients with end-stage renal disease. The most important challenge in PD therapy is the preservation of the peritoneum, since chronic exposure to glucose-containing PD fluids (PDFs) trigger inflammation, neoangiogenesis and fibrosis.

 

 

 

 

 

In Vivo Model

Mice were treated daily with peritoneal dialysis fluid (PDF) containing 4.25% glucose via a subcutaneously implanted mini-access port. Follow up is 5 weeks.

Available Readouts

  1. Ultrafiltration capacity of the peritoneal membrane

  2. ​​Structural changes of the peritoneal membrane

    • Masson Trichrome stain​

  3. Fibrosis

    • Picrosirius red staining specific for collagen I and III

    • Immunohistochemistry for fibronectin, a-SMA, collagen IV

    • Expression of profibrotic mediators

  4. Inflammation

    • proinflammatory mediators

    • F4/80+ macrophages

  5. Epithelial-Mesenchymal Transition (EMT)

    • Cytokeratin expression

  6. Angiogenesis

    • CD31, VEGF​

  7. Immunohistochemistry for specific markers of interests

  8. Apoptosis

Need further Readouts or cell lines? We routinely develop new models to fit our cutomer needs.

 

Speak with us for tailor-made solutions!

 

 

Techniques

  1. Histology (H&E, Masson Trichrome, Periodic Acid Schiff (PAS) staining, Picro Sirius Red staining for collagen I/III)

  2. Immunohistochemistry

  3. Flow cytometry including Bead-based flow cytometry

  4. ELISA

  5. Molecular biology (Real-Time PCR, Western blot)

In Vitro Models

1. Mouse peritoneal mesothelial cells (MPMC)

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Available Readouts

  • Pro-inflammatory mediator release

  • Epithelial-Mesenchymal Transition (EMT)

  • Apoptosis

  • Signal Transduction

2. Macrophages

Available Readouts

  • M1/M2 Polarisation

  • Pro-inflammatory mediator release

  • Apoptosis

  • Signal Transduction

Phenos - your independent contract research organisation

Contact us today. We routinely develop new models to fit your needs.

Image by National Cancer Institute

Renal Function after IR-Injury

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