The Effect Of Cord Blood Stem Cell Transplantation Combined With Acupuncture Therapy On The Expression Of The Brain-derived Neurotrophic Factor In Spinal Cord Injury

ObjectivesTo observe The effect of cord blood stem cell transplantation combined with acupuncture therapy on the expression of the brain-derived neurotrophic factor in spinal cord injuryMethodsA stable rat model of spinal cord T10 transection was established, ninety SD rats were divided into five groups by randon,normal group?injury model group?electropuncture group?umbilical cord blood group?combination group,each group is divided into three periods(7days?14days?28days),a total of 18 rats in each group .Separation spinal cord tissue from the rat each groups in 7days? 14days?28days, the application of immunohistochemistry,immunofluorescence,western bloting,RT-PCR technology Positioning and quantitative detection the expression change of neurotrophic factors (BDNF)in each organization, discuss the role of neurotrophic factors in combination therapy.Results1 The spinal cord injury model established in the early stage of the experimental group has a strong repeatability, which ensures the authenticity of the experimental data.2 To detect the expression of BDNF in gray matter at different time points after spinal cord injury in rats by immunohistochemistry: BDNF positive signal was light brown, mainly in the anterior horn of the spinal gray matter, almost all of which are located in the cytoplasm of neurons?Cell BDNF expression at different time points:(1) 7d,BDNF protein expression was significantly increased in the control group,transplantation group and combined spinal cord anterior horn motor neurons.; The transplantation group compared with the control group, the combination group compared with the transplantation group, positive signals in the spinal gray matter neurons were stained deeper, The expression level of BDNF protein was up-regulated.(2) 14d,The expression of BDNF in anterior horn motor neurons was increased in all groups, the number of positive cells was significantly increased, the cell body increased and the cytoplasm was stained deeply, there was a significant difference(P<0.05).(3) 28d,BDNF positive levels of control group and transplantation group were similar to the same group 14d time point,no significant difference(P>0.05) ; the expression level of BDNF in the combined group was further improved., express the peak to postpone?3 Immunofluorescence assay show: BDNF positive cells stained red, clear outline, the positive cells mainly were the astrocytes in the spinal cord?28 days after mode ling, compared with the control group rats, The fluorescence intensity of BDNF in the spinal cord of PD rats increased by 62.7%, 49.1% and 145% respectively(P <0.05)?Detection results showed: the expression of BDNF was highest in the combined group.4 PCR Real-time results show: BDNF expression in spinal cord tissue of 7d group rats with spinal cord injury was 9.8 ±1.0?8.0 ±1.5?22.2 ±3.9, compared with the model group(2.4± 0.8), the control group was significantly higher(Fig. 2A, B), the difference was statistically significant (P <0. 05)?The expression of 14d group were 3.8 ±0. 7?7.9.0 ± 1.3?13.2 ±1.6, compared with the model group((0. 4±0.05)),the control group was significantly higher,the difference was statistically significant (P <0. 05); The expression of 28d group were 3.4 ±0. 7?2.2±0.4?5.2±1.0,compared with the model group(((0.4±0.07)),the control group was significantly higher, the difference was statistically significant(P <0.05)?PCR Real-time detection results found: BDNF mRNA lower expression in model group, the expression of the treatment group was significantly increased?BDNF obviously increased after 7d damage, and reach the peak, 14d and 28d expression decreased, but it was significantly higher than the model group.5 Blot Western- results show: Compared with model group after modeling 28d,the protein expression of BDNF in SCI rat spinal cord tissue was increased 41.3%?3.5% ? 59.8% (P <0.05), the difference was statistically significant.Electric acupuncture treatment group, umbilical cord blood transplantation group and combined group expression of neurotrophic factor model group were higher than injury model group, umbilical cord blood stem cell transplantation with cupping therapy group expressed the highest amount.ConclusionsUmbilical cord blood stem cell transplantation combined with acupuncture can promote recovery of the function of spinal cord injury. After spinal cord injury,transplanted umbilical cord blood stem cell can survive, proliferation and migration;Umbilical cord blood stem cells combined with electro acupuncture treatment can promote spinal cord injury rats transplanted umbilical cord blood stem cell survival and proliferation, promoting the differentiation of umbilical cord blood stem cell into neurons and astrocytes, and could inhibit the differentiation of umbilical cord blood stem cell into astrocytes.