| Purpose: Through the observation of the urine flow dynamics related biochemical indicators combined with blood and urine,urinary bladder after spinal cord injury and to investigate the pathological changes of the kidney in rats and Placenta-derived mesenchymal stem cells(PDMSCs)derived neural cells transplantation on the recovery mechanism of neurogenic bladder function after spinal cord injury;the expression of Aquaporin 4(AQP4)in spinal cord injury and to explore its possible mechanism in spinal cord injury and the timing of cell transplantation for the treatment of pathological and physiological process,regulate the expression of Aquaporin 4(AQP4)in order to find effective methods or means of delay or block the neurogenic bladder after spinal cord injury.Method: The healthy adult rats 36,according to the SCI 1D before operation,after SCI 6h,1D,3D,5D and 7d point were randomly divided into 6 groups,6 rats in each group,T10 model of spinal cord injury by modified Allen 's method,the T10 segment of the spinal cord spinal cord damage detection of moisture content,the expression of AQP4 section T10 the RT-PCR of rats was detected;urodynamic test 60 female rats without exception after the rats were randomly divided into 3 groups: control group(A group,sham operation group,20);SCI group(group B,20),SCI+PDMSCs group(group C,20 rats);SCI.With the modified Allen' s method against T10 segment;cell transplantation in group SCI after operation 7d.The dynamics of rats in each group were measured respectively at SCI 1D before operation,after SCI 7d,14 d,30d,60 d point for urine flow,urodynamic test before collecting urine NAG detection,postoperative apical serum blood urea nitrogen BUN,SCI and 30 d after operation for bladder and kidney tissue of rats HE staining was performed to observe the morphological changes.Result:the animal model of survival: for AQP4 study of rats died 4,respectively,the time re modeling were added;for rat urine flow dynamics research group B 2 rats died in group C,3 rats died,respectively,the time to be re modeling based on.RT-PCR to detect the expression of AQP4 in spinal cord and the change of water content: the damaged segment T10,AQP4 in m RNA after SCI 6h expression was observed higher than that before operation,and progress over time increases in SCI and 3D after operation before reaching the peak,then the expression declined,until after the 7d is SCI slightly higher than before surgery;after spinal cord injury 6h observed hit segments of T10 spinal cord increased water content(edema),SCI and 3D after operation of spinal cord edema reached the peak,then the degree of edema reduced to 7d after injury is still slightly higher than SCI before operation,but as time continues,the degree of spinal cord edema(including spinal cord falling slowly,but water)to reduce the degree of edema.Urodynamic examination: compared with the control group,after SCI 7d,the bladder capacity and residual urine of the rats in the simple injury group were significantly increased than those of the control group rats,and with the progress of time they increased at first and then decreased.After SCI 60 d,the bladder capacity of the rats in the simple injury group was still significantly higher than that of control group rats,the differences between the two groups are statistical significance(P<0.05).The urine volume of rats decreased at first and then increased and then decreased,after SCI 14 d to 60 d the urine volume was significantly higher than that of the control group rats,but the micturition efficiency was significantly lower than that of the control group,the difference is statistically significant(P<0.05).The rats in cell transplantation group after SCI 30 d,the bladder capacity and residual urine volume of the rats were significantly lower than those of rats in the simple injury group,but urine volume and micturition efficiency of the rats in cell transplantation were significantly higher than those of the rats in simple injury group,the difference was statistically significant(P<0.05).Morphological observation: Gross kidney specimen: kidney specimens of rats had no obvious increase in size,cut the kidney longitudinally along the long axis and saw the spinal cord injury group,compared with the control group,the separation of the ureter of the rats in the SCI+PDMSCs cell transplantation group was slightly larger.Three groups had no obvious kidney hydrocele phenomenon.Gross specimens of bladder: the bladder volume of the rats in the spinal cord injury group and the SCI+PDMSCs cell transplantation group,in which the spinal cord injury group has the maximum bladder volume.cut bladder layers along the longitudinal axis of the bladder could see that the bladder walls of the spinal cord injury group and the SCI+PDMSCs cell transplantation became thinner than those in the control group.The thickness of both groups has no significantly megascopic difference.HE staining of kidney tissue morphology Results: Control group: lightly stained epithelial cells of the renal tubular,the size of nucleus was normal,the basement membrane remained intactness and continuousness.Spinal cord injury group and spinal cord injury+PDMSCs cell transplantation group: At the early time,renal pelvis wall and renal interstitial wall appear lymphocyte-predominant inflammatory infiltration,commonly appeared in the deep medulla and with time progress it get severer.Tubular epithelial occurred damage,the necrotic epithelial cells concentrated,and the cell nucleus got concentrated and anachromasis,the basement membrane got integrally and continuously damaged.Renal interstitial appeared focal distribution of fibrosis,the glomerular morphology was normal and it had no significant atrophy and fibrosis.In later period,the impaired renal tubular epithelial cells occured regeneration,but they were not arranged in the basement law,they had the performance of large and small,and disorganized form.Bladder HE stained tissue morphology Results: Compared with the control group of rats,the bladder tissue of rats in spinal cord injury group all appeared a large number of inflammatory cell infiltration and vacuolar changes.The epithelial cells showed varying degrees of disorder,in severe case,it appeared exfoliation changes.In spinal cord injury + PDMSCs cell transplantation group also had the above performance,but it had slighter degree of the performance than that of simple injury group.Light microscopy and PDMSCs in supernatant of rat spinal cord ho mogenate after induction: primary PDMSCs were round or nearly round,s mall volume,mixed in red blood cells is not easy to observe after cultured th ird generations,the cells were spindle shaped and th volume significantly in creased comparatively,cells showed colony or cluster growth;join spinal cord homogenate supernatant after induction,cells were neuron like structure,cell processes increased and tapering,manifested as bipolar or multipolar cel l like structures,neural stem cell specific marker Musashi was detected by in duction,PDMSCs cells showed positive expression.The detection of the biochemical index :it reflects blood urea nitrogen(BU N)in glomerular function having no apparent change 1d before SCI and 60 d after SCI modeling,the difference was not statistically significant(P>0.05);renal tubular injury marker urinary enzymes NAG is elevated in the early stage of spinal cord injury,and gradually increased with the time progression;urinary enzyme NAG detection but the cell transplant ation group rats after 60 d.Its value was significantly lower than that of the s imple injury group,the difference was statistically significant(P<0.05).Conclusion:1.PDMSCs nerve cell transplantation can improve the bladder function in ra ts with spinal cord injury.2.PDMSCs nerve cell transplantation can repair the renal function of rats wit h spinal cord injury.3.Renal tubular interstitial changes in the evaluation of spinal cord injury aft er neurogenic bladder caused by renal dysfunction than the glomerular patho logical changes are more valuable. |
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