| To investigate the Effects of intraperitoneal injection of valproic acid(VPA)combined with transplantation of bone marrow mesenchymal stem cells(BMSCs)on neurological function recovery after spinal cord injury(SCI)in rats.BMSCs from 3-4 weeks old healthy male Sprague-Dawley(SD)rats were cultured in vitro.The third generation BMSCs were detected by flow cytometry and collected for use.Sixty adult male SD rats were randomly divided into five groups: Sham operation group,model group,valproic acid group,transplantation group and combined group.Spinal cord injury model was maded by modified Allen's technique.Valproic acid group was given intraperitoneal injection of VPA(300 mg?kg-1)after injury,then repeated per 12 h until killing.At transplantation group,1 mL BMSCs,it's concentration is 1×106 mL-1,were injected into the spinal cord of injured segment.While to combined group,both VPA and BMSCs were injected.In addition,Sham operation group and model group received the same dose of normal saline at the same time point.The recovery of the locomotor function of each group was evaluated with BBB scale at the 7 d and 14 d after injury,and then rats were sacrificed.The area of syringomyelia was observed by paraffin section and HE staining.And the expression of Caspase-3 protein was detected by immunohistochemistry.Then the neuronal apoptosis was observed by TUNEL staining.BBB Rating: On the 7 d after SCI,the rate in sham operation group,model group,valproic acid group,transplantation group and combined group were(21),(2.22±0.33),(3.80±0.53),(3.81±0.83),(4.02±0.83).On the 14 d after SCI,the rate in five groups were(21),(4.51±0.41),(7.25±0.95),(7.32±0.55),(9.76±0.95).The motor function of sham operation group was not affected.The BBB score of the three treatment groups was significantly higher than that of model group(P<0.05).The score of combined group was significantly higher than that of group valproic acid group and transplantation group on the 14 d after SCI(P<0.05).HE staining results showed the area of the lesion area of model group,valproic acid group,transplantation group and combined group were respectively(4.57±0.26)mm2?(3.34±0.21)mm2?(3.51±0.18)mm2 and(2.43±0.35)mm2.Combined group was significantly lower than the other three groups(P<0.05).Immunohistochemical staining: On the 7 d after SCI,the caspase-3 expression in sham operation group,model group,valproic acid group,transplantation group and combined group were(1.51±0.23)%,(28.51±5.23)%,(24.39±4.65)%,(24.51±3.81)%,(20.73±3.43)%.On the 14 d after SCI,the caspase-3 expression were(1.64±0.41)%,(26.22±4.81)%,(21.21±4..04)%,(21.47±3.32)%,(16.15±3.09)%.Immunohistochemical staining showed that a small amount of caspase-3 expression was found in sham operation group.The expression of caspase-3 in three treatment groups was significantly lower than that model group(P<0.05).While the expression of caspase-3 in combined group was significantly lower than that in valproic acid group and transplantation group(P<0.05).Apoptosis results: On the 14 d after SCI,the apoptotic index in sham operation group,model group,valproic acid group,transplantation group and combined group were(4.18±0.39)%,(40.51±3.81)%,(19.95±2..04)%,(20.87±2.32)%,(12.83±1.09)%.A small number of apoptosis-positive cells were found in sham operation group on the 14 d after injury.Compared with model group,the apoptosis index of the three treatment groups was lower(P<0.05).The apoptosis index of combined group was significantly lower than that of valproic acid group and transplantation group(P<0.05).It is very significant,Effects of intraperitoneal injection of valproic acid combined with transplantation of bone marrow mesenchymal stem cells on neurological function recovery after spinal cord injury in rats.The mechanism may be related to down-regulation of caspase-3 expression,leading to inhibition of apoptosis. |
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