Mouse Model for Renal Fibrosis (RF)

1. 3% sodium pentobarbital (80mg/kg)intraperitoneal injection of anesthesia, remove the left back hair, skin disinfection.
2. Cut at 1cm of the left side of the spine on the top of the thigh, and cut the skin and muscles.
3. Separated the left ureter, dissociation of ureters at the 1/3 of the ureter and ligate twice with 6-0 at different site, and then cut ureter between the ligations, clean up the wound and make the two-layer suture. After the mice awake, put them back to clean cage, observe the state and death of mice, and make records.
4.kill the mice on 14d, 7d, 21d, 28d, respectively,take the left part of the kidney tissue, 4% poly formaldehyde solution fixed, paraffin embedded; the rest of the kidney tissue liquid nitrogen freezing and stored at -80℃.

1.Urine protein and Urine NAG activity determination
Urinary protein and urinary NAG activity of the model group is significantly higher than that of the normal group and the sham operation group at 2 and 4 weeks after operation.
2. Serum creatinine (Scr), Urea nitrogen (BUN) content is determined after 4 weeks of operation, the model group serum Scr and BUN levels is increased.

The left kidney is fixed with 4% poly formaldehyde, dehydrated, transparent, paraffin embedded and sliced. By HE staining and Masson staining, the pathological changes and inflammatory cell infiltration of glomerular and renal tubular interstitial are observed by light microscope.
In model group mice renal: interstitium inflammatory cell infiltration, renal tubular epithelial cell atrophy, loss of brush border, the tubular lumen expansion cystic, lumina collapse, interstitial widened obviously, interstitial visible inflammatory cell infiltration, accompanied by more into fibroblast proliferation. The inflammatory cell infiltration increased with time, the tubular epithelial cells were flat and detached, and the renal tubular atrophy, dilated, even collapsed, disappeared, collagen fiber hyperplasia, interstitial fibrosis increased.

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.