| ObjectivesNeurogenic bladder(NB)is a kind of urinary tract impairment due to the disorders and dysfunctions of neural urination control system.There are neural diseases or trauma in the nervous system.Because of the injuries in different nerve segments,the symptoms of neurogenic bladder are not the same.The complications of neurogenic bladder after spinal cord injury mainly are urinary tract infection(UTI),function deterioration of upper urinary tract and low urinary tract,urinary system stones and so on.Particularly,the worst complication in neurogenic bladder is kidney failure.Now,the neurogenic bladder research is insufficient in the world,and no golden standard treatments.These are a subpopulation of stem cells that can be easily isolated and proliferated form human voided urine,i.e.urine derived stem cells(USCs).These cells display the same characteristics of other stem cells and are capable of self-renewing and differentiating.And USCs are much easier to be obtained than other stem cells.They avoid the problems of ethics.Because USCs come form the urinary tract system,they have better homogeneity and homology.USCs have achieved significant effects in tissue engineering repair and treatments of urinary system.To observe the function and histological changes of neurogenic bladder,and to investigate the effect of Urine-derived stem cells transplantation via tail vein injection on the neurogenic bladder after spinal cord injury(SCI).USCs provide the new treatment idea and strategy in neurogenic bladder.Methods1.Establish the neurogenic bladder models: the female Sprague-Dawley rats(180-200g)are applied in this experiment.Rats were randomly divided into the normal group,the SCI-induced group,the SCI-induced and Urine-derived stem cells transplantation group in the study.On 7 days after induction of SCI,inject the stem cells transplantation group rats with Urine-derived stem cells via tail vein.2.Western Blot: 28 days after the injection of stem cells,we would extract the total protein from bladder.The SDS-PAGE agarose gel electrophoresis would be used to seperate the total protein which would be kept in 5% milk afterwards.The P2Y4 polyclonal antibodies’ incubation will begin in the sealed 5% milk.After the incubation of the secondary antibody,the Image system would be adopted to measure.3.Histological detection: weigh out the bladder wet weight immediately after extract the bladder tissue,embed the bladder tissue into paraffin wax,section it and adopt H.E.staining and TUNEL staining.4.We use the Urinary dynamic study to measure the functional difference between bladders of rats from different groups under anaesthetization.The Bladder muscle strips tension measurement is employed to measure the different comtraction of bladders after rats die.Results1.After adopting the Western Blot measurement,it showed that the P2Y4 protein is obviously higher than that of normal group rats and stem cell group rats.2.The SCI-induced and Urine-derived stem cells transplantation group is better in bladder wet weight(0.31±0.01 g),bladder wet weight/body weight(1.19±0.41),when compared with the SCI-induced group(0.66±0.07,2.30±0.26).3.In SCI-induced group,H.E.staining showed that the bladder had the serious edema and the detrusor structure was disordered.But the SCI-induced and Urine-derived stem cells transplantation group bladder improved the edema and thickening of detrusor.TUNEL staining showed the SCI-induced group bladder muscle cell apoptosis rate(59.80±14.18)was much higher than the the normal group(8.20±3.13%)and the SCI-induced and Urine-derived stem cells transplantation group(59.80±14.18).4.After the Urinary dynamic study measurement,it demonstrated that the contraction time and urinary output of the stem cell rats are noticeably improved when compared with those of the induced group rats(P<0.01)whereas the bladder basic pressure and the bladder peak pressure were not obviously improved(P>0.05).Conclusions: In the establishment of neurogenic bladder models,urine derived stem cells could promote the recovery of the bladder functions,relieve the edemas and bladder’s hyertrophy as well as improve the urination. |
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