| Background:Ureteral obstruction is very common in Urology. It will result in irreversible pathophysiology of renal interstitial fibrosis and finally develop into end-stage renal failure without treatment in time as well as other chronic kidney diseases. Unilateral ureteral obstruction (UUO) has been used as an ideal model for the study of the mechanisms of renal interstitial fibrosis (RIF) and chronic kidney diseases in animal experiments.In recent years, study on therapy of renal disease using mesenchymal stem cells (MSCs) has made great progress. MSCs have shown a wide prospects of clinical application, however, the dynamic distribution and effect of MSCs confuses many researchers.In vivo imaging system (IVIS) is used to monitor the track of labeled MSCs in the same animal through the whole experiment. Therefore, the death number of experimental animals is reduced at each time point, but the results is more credible.Objective:I used an animal model of reversible unilateral ureteral obstruction (R-UUO) to simulate clinical ureteral obstruction which received operation. MSCs labeled with luciferase and GFP were transplanted into the R-UUO animals through renal artery. Homing of MSCs was evaluated by IVIS. Finally MSCs-induced effect on obstruction-induced RIF was evaluated by immunohistochemistry and real-time quantitative PCR. The results may be useful for later research and clinical practice.Innovations:1. There has not been any research on the evaluation of arterially delivered MSCs-induced effect on obstruction-induced renal interstitial fibrosis;2. Tracking the homing of MSCs using IVIS in R-UUO animal model is an innovative method.Methods:1. Thirty-six SPF femal BALB/c mice (4W) were divided into three groups:Sham, PBS and MSCs. Every group contained six mice at each time point;2. Transfected MSCs with luciferase gene and GFP gene, and subcultured the Luc-GFP-MSCs;3. Built up the animal model of R-UUO with a nontraumatic microvascular clip in the PBS and MSCs groups. The left ureter was only isolated but not clamped in Sham group;4. After reversal of obstruction,1×106labeled MSCs were transplanted into the R-UUO animals through renal artery in MSCs group. Sham group just received the same volume (50μl) of phosphate buffered saline;5. Three groups of mice were imaged by IVIS at the1st,3rd,7th,14th and28th day after transplantation. Frozen sections were made to prove the homing result at the3rd and7th day additionally;6. The expression of E-cadherin, a-SMA, TGF-β1, and TNF-a in three groups was measured by real-time quantitative PCR. The data was analyzed by SPSS18.0statistical software.Results:1. The character of MSCs was not affected by transfection and subculturing. The light intensity increased with the number of Luc-GFP-MSCs;2. I successfully set up the mice R-UUO model, and improved the stability and equilibrium of the model;3. I built up and perfected the technique of delivery route via renal artery;4. There was no specific photon in Sham and PBS group. The photons diffused at left kidney site in MSCs group, and the luminescence density of the kidney site increased steeply from one day to three days and then moderately decreased with time but could not be observed for the14th day.5. There was no statistically significant between PBS and MSCs group at the1st week (P>0.05), but they were different with Sham group (P<0.01). The result of TGF-β1expression at the4th week was similar to that at the1th week. But the E-cadherin, a-SMA, and TNF-a expression of three groups were different significantly (P<0.01)Conclusions:1. MSCs have the character of homing to obstructive kidney;2. MSCs transplanted through the renal artery attenuate obstruction-induced RIF. |
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