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Mouse Model for Renal Ischemia-Reperfusion Injury (RIRI) Mus musculus (Mouse) Disease model

Renal Ischemia

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Overview
Properties
  • Prototype SpeciesHuman
  • SourceIschemia-Reperfusion, Clamp of Bilateral renal artery
  • Model Animal StrainsBalb/c mice (SPF level), male, week age:4w~6w, body weight:20g~22g.
  • Modeling GroupingRandomly divided into groups: Control group, Model group, Positive drug group and Test drug group (three concentration gradient group), 15 mice per group.
  • Modeling Period24h、72h
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  • Mouse Model for Renal Ischemia-Reperfusion Injury (RIRI) Packages (Simulation)
  • Mouse Model for Renal Ischemia-Reperfusion Injury (RIRI) Packages (Simulation)
  • DSI529Mu01.png Fig 1.Clamp of bilateral renal artery,ischemia 45 mins and loose
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Modeling Method

1. 20-22g mice, fasting 12h, free drinking water before operation.
2. 3% sodium pentobarbital (80 mg/kg) intraperitoneal injection of anesthesia, shave back, skin disinfection.
3. Cut the skin and muscles in the lower back 0.5cm, the lower edge of the rib 0.5cm, visible to the kidneys, carefully separated from both sides of the renal artery of the kidney, the rapid use of arterial clamp on both sides of the renal artery.
4. Ischemia 45 mins, loosen the artery clamp, restore blood flow, observe the renal recovery.
5. Suture, after the mice awake, put it back to clean cage, observe the state and death of mice, and make records.
6. Control group do not take the ischemic operation, the other operation is the same.
7. Take samples at 0h, 3h, 6h, 12h, 24h, 72h six time points after reperfusion. Anesthesia mice, pick the eye to take blood, 4℃,
3000r, 10 minutes to get serum, stored at -80℃. At the same time, take the left kidney tissue to be used as samples.

Model evaluation

1. Detection of serum biochemical indexes:
Take (0h, 1h, 3h, 6h, 12h, 24h, 72h) serum, detection of serum BUN (urea nitrogen) and Scr (serum creatinine) level, to assess renal function.
2. Detection of renal coefficient:
The removal of both kidneys, physiological saline, weighing calculation renal coefficient.
Renal coefficient = bilateral kidney weight (mg)/body weight (g)
3.Tubular necrosis score index:
Each section x 200 times mirror the outer medulla 10 horizons,
0 = normal,
1 = minor damage (impaired tubular < 5%),
2 = mild damage (renal tubular damage 5% ~ 25%),
3 = moderate injury (damaged renal tubular 25% ~ 75%),
4 = severe injury (damaged renal tubular > 75%) for semi quantitative analysis and calculate the mean.
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Pathological results

4% poly formaldehyde solution fixed for 48h. Conventional tissue dehydration, transparency, paraffin wax, embedding. Paraffin sections are stained with HE and PAS.
In control group, the glomerular, renal tubular and interstitial structure of the mice are normal.
In the model group, with reperfusion time extended, presenting different degrees of pathological changes, in renal tubular epithelial cells cloudy swelling, water or vacuolar degeneration, loss of brush border, part of the solidification of renal tubular epithelial cell necrosis, fall off, can be seen within the lumen of the tube type, and visible interstitial edema, interstitial focal infiltration of inflammatory cells, glomerular lesions is not obvious.

Cytokines level

ELISA methods are used to detect serum levels of IL-6 and TNF-α. The content of TNF-α and IL-6 in model group was significantly higher than that in control group at all time points, and 24h was the most significant.

Statistical analysis

SPSS software is used for statistical analysis, measurement data to mean ± standard deviation (x ±s), using t test and single factor analysis of variance for group comparison , P<0.05 indicates there was a significant difference, P<0.01 indicates there are very significant differences.

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