【Animal Modeling】-Oleic acid induced acute renal failure model in rats

  [Model mechanism] Oleic acid is a highly toxic fatty acid. When oleic acid is injected into the left renal artery, renal tubular epithelial cells swell and necrosis, interstitial edema, intravascular renal ischemia and severe renal microcirculation. It is impaired and renal excretion function cannot form AFR.

  [Modeling method]

  1. Wistar male rats weigh 180-250 grams. Oleic acid, bromobenzene, triphenyltetrazolium chloride (Trc). The rats were randomly divided into 5 groups, the normal saline control group (left renal artery normal saline injection 0.45ml/kg), oleic acid group I, oleic acid group II, and oleic acid group. III, oleic acid group IV, rats in the experimental group were injected with 0.05 ml/kg, 0.1 ml/kg, 0.125 ml/kg, 0.15 ml/kg oleic acid in the left renal artery. To ensure the correct dose of oleic acid during injection, please dilute oleic acid with vegetable oil. The results before the test showed that there was no significant difference between the vegetable oil group (0.45 ml/kg) and the saline group.

  2. Oleic acid was injected into the left renal artery at 10 minutes, 6 hours and 24 hours, and the changes in the kidney TTC staining were observed using TTC staining. Routinely prepare electron microscope and optical microscope sections, and observe the ultrastructure of renal tubules and capillaries around the renal tubules through a 400T transmission electron microscope. Under an optical microscope, 10 high-power fields were observed for each section. At the same time, renal tubular necrosis and tubular casting were 0/HP (-), 1-2/HP (+), 3-4 / respectively. HP means (++), → 4/HP means (+++), and the right kidney, lung, heart, brain, liver, small intestine and testis can also be observed under an optical microscope.

  3. 10 minutes, 1 hour and 6 hours after the injection of oleic acid in the left renal artery, the changes of the left renal plethysmogram were recorded by impedance method. BUN and Cr were measured 24 hours after injection of oleic acid.

  [Model Features] 1. After infusion of oleic acid, renal microcirculation is severely damaged, glomerular filtration, renal tubular secretion and reabsorption are all damaged, leading to impaired renal excretion. Ten minutes after the injection of 0.15 ml/kg oleic acid into the left renal artery, the glomerular capillary endothelial cells were covered, the nuclear chromatin was highly aggregated, and the nuclear membrane partially disappeared. , Mitochondria are swollen, and the basement membrane is locally thickened. Six hours after injection, the above-mentioned changes intensified, and endothelial cell nuclear fragmentation was observed, and in severe cases, endothelial cells were destroyed. Twenty-four hours after the injection of oleic acid, the nuclei of glomerular capillary endothelial cells contracted, the nuclei divided and dissolved, the endothelial cells collapsed, and the basement membrane was destroyed.

  2. Kidney histology changes are related to the injection dose of oleic acid. In the oleic acid I-IV group, renal tubular necrosis was observed. In the oleic acid II-IV group, the glomeruli were swollen and even the glomerular nuclei disappeared, the cytoplasm was stained red, the structure was destroyed, and the oleic acid group was destroyed . The degree of injection increased. The left kidney becomes pale after TTC color development, which is related to the action time of oleic acid and varies with the dose of oleic acid, but the right kidney of each group is bright red. The chromogenic and positive patients showed non-cortical, light-colored, and light-colored areas, mainly distributed in the cortex and extramedullary. In severe cases, the entire kidney is involved.

  3. 10 minutes, 1 hour and 6 hours after injection of oleic acid, renal impedance pulse waves can be seen. The wave heights of flat wave and flat wave were 34%, 32% and 32%, respectively, which was 65% lower than the control group. 68% and 66% of the people respectively showed a decrease in the elasticity of the renal artery, an increase in tension, an increase in peripheral resistance, and a significant decrease in blood perfusion of the kidney tissue. The serum BUN level depends on the oleic acid injection dose. Oleic acid in group IV was significantly higher than that in other groups. There was no significant difference in serum Cr between the two groups.

  [Model Evaluation and Application] This model has severe renal microcirculation disorders accompanied by interstitial and parenchymal damage. It can be used to simulate human ARF caused by renal microcirculation disorders.. There are five most commonly used ARF experimental animal models. The renal artery was clamped, norepinephrine was injected directly into the renal artery, glycerol was injected intramuscularly, and mercury chloride and uranium nitrate were injected intravenously. These models are mainly used for the study of parenchymal cells. In this model, infusion of oleic acid through the left renal artery can cause renal microcirculation and parenchymal cell damage. The established rat ARF model was compared with the above five models. This model is one of unilateral renal microcirculation disorders with severe renal parenchymal damage. The new ARF model can be used to study the cause and treatment of acute renal failure.