| Objective: To investigate the efficacy of marrow stromal cells (MSCs) grafted on to the regenerating spinal cord of injured rats and also measure the change of basic fibroblast growth factor (bFGF). We hope to provide evidence that MSCs enhances the expression of endogenous bFGF which is needed in spinal cord injured rats.Methods : Animal spinal cord injured (the T12 spinal cords segment) model was established by modified Allen's WD method. 66 healthy adult srague-dawely (SD) female rats, mean weight 200-250g,were randomly divided into eleven groups: one control group ; spinal cord injury(after operation 1st,3rd,7th,14th,21th day); and MSCs transplantation groups(Two days after SCI, the injured spinal cords were injected with MSCs suspension),(after operation 1st,3rd,7th,14th,21st day)。The fluorescent cells were observed in spinal cord with fluorescent microscope. Then,anti-bFGF serum were applied wuth the immunohistochemical ABC method to detect the distribution and expression of bFGF in spinal cord'tissues at different period time after SCI and GFP-MSCs transplantation , and were analysed by quantitative grey density.Result: 1．The GFP-MSCs can be found in the spinal cord injured sections of all GFP-MSCs transplantation groups. The GFP-MSCs appeared as green fluorescent dots. GFP-MSCs were aggregated together in the location of the injured spinal cord on the 1st day, so it was difficult to distinguish between individual cells. By day 3 the cells began to spread out. By day 7, the cells were easy to distinguish and uniformly distributed at the center of damage. Day 14 was the same as day 7．By day 21 the cells began to migrate uniformly and distantly.2．The result of expression of bFGF : In the normal control groups,spinal cord injured groups and GFP-MSCs transplanted groups, the bFGF immunoreaction product were mostly located in the cytoplasm of neurons and neural glial cells. In the injured groups and GFP-MSCs transplanted groups, the number of bFGF positive nerve cells began to increase on the 1st day group, came to a climax with the 7th day group and by the 21 st day group decreased to normal group level. The bFGF positive nerve cell on the 3rd,7th,and 14th day spinal cord injured groups and the 3rd,7th,14th,and 21st day GFP-MSCs transplanted groups were more than the normal group(p<0．05) .The 1st day,21st day spinal cord injured groups and 1st day GFP-MSCs group closed normal group (P>0．05) ; while in the 3rd day,7th day,14th day,21th day GFP-MSCs transplanted groups showed significant difference compared with that spinal cord injured groups(p<0．05) .The average grey degree value firstly appeared reducing,then Increasing.The spinal cord injury and cell transplantation groups were lower than the control group (p <0．05) . Cell transplantation in the same period were lower than the spinal cord injury.3．The result of motor function scorce: the BBB score of spinal cord injured control groups and GFP-MSCs transplanted groups were lower than normal control group(p<0．05) ,there no significant difference between the 1st day,3rd day,7th day GFP-MSCs transplanted groups and spinal cord injured groups(p>0．05) ,but 14th day,21th day of the two groups have big difference(p<0．05) .Conclusion: 1．The different expression tendency of bFGF after SCI suggests that bFGF may play important role in rats. 2．The result of BBB score indicated that MSCs could improve the recovery of motor function of spinal cord injury in the rat model. 3．Changeing in the expression of bFGF after spinal cord injury, suggesting their protection in the neurons of the spinal cord injury .4．After transplantation of MSCs enhance expression of endogenous bFGF. Which suggests that MSCs participate in the repair of spinal cord injury. Abjective: Aim To explore the inhibitory effect of dexamethasone on expression of TNF-a and IL-6 in spinal cord injury.Method: 90 healthy adult srague-dawely(SD) rats, mean weigh 220-250g, were divided into 15 groups including one normal control group,contused spinal cord injuried control seven groups(1hour group,3hour group,6hour group,12hour group,24hour group,36hour group,48hour group)and dexamethasone to treat contused spinal cord injury seven groups(1hour group,3hour group,6hour group,12hour group,24hour group,36hour group,48hour group). Establish spinal cord injuried animal model: With 10gx5．0cm impact to the T12 spinal cord segment was established by modified Allen's WD method. Given Dexamethasone at the time of 15min after being made laminectomy and in SCI. Then fixed the spinal cord segements (caudally to the T12 spinal cord segments) and taken out from the rats with 4%formaldehydum polymerisatum (4℃), to put them into 15%,30%sucrose overnight .These tissues were made into frozen section in 20μm thickness. Then Hematoxylin and eosin staining. Lastly, anti-TNF-αand anti-IL-6 serum were applied wuth the immunohistochemical ABC method to detect the distribution and expression of TNF-αand IL-6 in spinal cord. HMIAS-2000 medical use of high-resolution color graphic analysis immune system detection of TNF-αand IL-6 The positive reaction of the average gray value. The result was analysis with the SPSS 11．0 software.Result: 1 .In spinal cord of control group ,TNF-αand IL-6-like immunoproduct was hardly any observed in tissue.2．The expression of TNF-αimmunoreaction product were observed after spinal cord injuried ,They were mostly located in plasma in the nucleus and neural glial cells.TNF-αimmunoreactive increased at 1hpo (hour post operation) and reached the peak at 24hpo,then decreased, returned to normal control group within 48hpo .Injected in dexamethasone after injuried spnal cord within 15 minutes, the immunoreactive product was still existed, then peaked at 6h.While crest of the wave is lower than before. It is gradually declining Since 12h,to 48h resumed normal. 3．The expression of IL-6 immunoreaction product were observed by the injuried spinal cord , They were mostly located in plasma in the nucleus and neural glial cells.IL-6 immunoreactive increased at 1hpo (hour post operation) and reached a peak at 12hpo,then decreased, to 48hpo closed normal control group. Injected in dexamethasone after injuried spnal cord within 15 minutes, the immunoreactive product was still existed, then peaked at 6h.While crest of the wave is lower than before. It is gradually declining Since 12h,to 48h resumed normal.Comclusion:1．TNF-αand IL-6-like immunoproduct dosen't exist in the spinal cord normal group of the rat.2．The expression of TNF-αand IL-6 immunoreaction product is appeared Immediately after spinal cord injuried, while is Shorter duration.It is disappeared to 48h. Which is suggested they participate in the inflammatory reaction after spinal cord injury.3．High-dose dexamethasone markedly inhibited in The expression of TNF-α, IL-6, which is guesses that not only as we know anti-inflammatory effects, but also protect nervous system action. |
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