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A novel in vivo renal ischemia reperfusion injury (IRI) model induced solely by cold ischemia
Author(s) -
Zhang Jie,
Wang Shaohui,
Wei Jin,
Wang Lei,
Zhang Gensheng,
Yip KayPong,
Liu Ruisheng
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1217.1
Subject(s) - medicine , ischemia , kidney , aorta , renal artery , inferior vena cava , abdominal aorta , renal vein , transplantation , renal ischemia , cardiology , anesthesia , reperfusion injury
Transplanted organs experience several episodes of IRI during procurement and transplantation in forms of both warm and cold ischemia. IRI‐induced kidney injury is crucial for the survival of transplanted kidney and considered as one of the most important nonspecific factor affecting graft function and is associated with decreased long‐term survival. We don't know whether they are the same or different of IRI induced between warm and cold ischemia, since no IRI in vivo model that is induced solely by cold ischemia available. In present study, we developed a novel IRI mouse model, which is induced exclusively by cold ischemia. C57BL/6 mice were anesthetized with pentobarbital (50 mg/kg i.p.) and body temperature was maintained at 37 °C during the surgery. Right renal artery, vein and ureter were ligated and right kidney was removed after a midline abdominal incision. Left kidney, aorta and inferior vena cava were exposed. The left kidney was placed in a kidney cup and incubated with cool saline, which temperature was maintained at 4±0.5°C by circulating sterilized saline solution from a reservoir on ice and driven with a pump. Four 5 mm microvascular mini clips were used to prevent bleeding and induce ischemia. First, the aorta between the left and right renal arteries was clamped, and distal aorta and inferior vena cava were clamped together. A small opening was cut on the left renal vein after clamping a short segment of left renal vein near vena cava. The aorta below left renal artery was infused with 1–1.5 mL of 4°C 0.9% saline using 30G needle until the solution from the renal vein became clear. The left renal pedicle between the opening and the left kidney was clamped to induce ischemia. The small opening was sutured with 10/0 suture. Then three clips on the aorta, inferior vena cava and left renal vein were removed. The clip on renal pedicle was removed 60 min after clamping and renal blood restored. The wounds were sutured and mice were recovered. The animals were anesthetized again 24 hours after IRI. Blood (500ul) was collected through retro‐orbital venous sinus before sacrifice and left kidneys were harvested and fixed in 10% formalin. Plasma creatinine level was significantly increased to 2.47±0.05 mg/dl compared with the control (0.08±0.03 mg/dl) and the plasma kidney injury molecule‐1 (KIM‐1) increased to 2917±492 pg/ml compared with the controls (2917±34 pg/ml). Histology with PAS staining showed that the most severe injury in corticomedullary region, with a percentage of tubular necrosis 31.4±8.1% and moderate damage in renal cortex, outer‐medulla and inner medulla with a percentage of tubular necrosis 1.6±1.3%, 8.9±4.7%, and 4.6±2.9%. In summary, we have developed a novel in vivo IRI mouse model that is exclusively induced by cold ischemia. This model could provide important information and mechanism for IRI that induced by cold ischemia in kidney transplantation.