HDAC1去乙酰化调控p11/tPA/BDNF信号通路在BMSC源性纳米支架移植治疗大鼠脊髓损伤的研究

The disability rate after spinal cord injury (SCI) is very high, while there is no effective method for treatment up to now. Stem cell transplantation has been considered as an important route to improve the nerve repair after SCI. In our previous studies, we found transplantation of mesenchymal stem cells (BMSCs) from transgenic mice in situ could improve the spinal cord function recovery after injury, but the effect is unsatisfactory. In the present study, we planned to transplant OBC nano stent, with BMSCs and lentivirus-mediated upregulation of histone deacetylase (HDAC1), into the rats’ transected spinal cord. Then, we will evaluate the effects of BMSCs-HDAC1 network stent transplantation by observing the regeneration of injured axons, the synaptic plasticity and recovery of myelin. Meanwhile, the changes of p11/tPA/BDNF signal pathway in the cord tissue will be detected to assess the route which the BMSCs-HDAC1 network stent functions in, especially the molecular mechanisms of p11 acetylation and deacetylation modification. In summary, the study was designed to reveal the possibility of HDAC1 involved in BMSCs-network stent transplanted effect of neural plasticity, and try to provide some new information and experimental basis for treatment of SCI.

脊髓损伤致残率极高，目前缺乏有效治疗手段，课题组前期证实转基因小鼠骨髓间充质干细胞（BMSC）原位移植可促进脊髓神经功能修复，但单一移植效果不理想。本研究拟分离GFP转基因小鼠BMSCs，慢病毒介导上调去乙酰化酶（HDAC1）表达后注入纳米支架，移植入脊髓全横断大鼠体内，检测BMSCs存活分化、凋亡等功能改变；运用透射电镜和BDA顺行追踪等方法，检测损伤神经元纤维再生、突触通路构筑和髓鞘修复情况，观察BMSCs-HDAC1源性神经网络支架移植治疗脊髓损伤的作用；通过调查损伤头、尾端脊髓中相关细胞因子，尤其p11/tPA/BDNF信号通路的表达变化，进一步验证p11去乙酰化后，下游通路蛋白表达改变，内源性BDNF活性强化，阐明HDAC1参与调控促进BMSCs源性纳米支架移植后脊髓运动、感觉神经功能可塑性修复。为临床靶向性神经调控+干细胞源性生物工程支架联合治疗脊髓损伤，提供新策略和实验数据。

脊髓损伤致残率极高，目前缺乏有效治疗手段，干细胞移植促进损伤修复是一重要的研究方向，课题组前期证实转基因小鼠骨髓间充质干细胞（BMSC）原位移植可促进脊髓损伤神经功能修复，进一步研究分离GFP转基因小鼠BMSCs，慢病毒介导上调去乙酰化酶（HDAC1）表达后，注入纤维素支架，移植脊髓全横断SD大鼠体内，检测BMSCs存活、分化、凋亡等细胞功能改变；运用透射电镜和BDA顺行追踪等方法，检测损伤神经元纤维再生、突触通路构筑和髓鞘修复情况，观察BMSCs-HDAC1源性神经网络支架移植治疗脊髓损伤的作用；通过调查损伤处头、尾端脊髓中各种相关细胞因子，尤其BDNF信号通路的表达变化，证实可促进损伤脊髓运动、感觉神经功能的可塑性修复。在此基础上，我们拓展研究，通过大数据分析，高通基因编辑和单细胞测序、全组织透明成像技术，探讨小鼠视神经或脊髓损伤后，小胶质细胞等在增殖静默和胶质反应等不利因素下，设计多组学整合研究思路，摸索其转变为干细胞，进一步重编程为神经元或神经节细胞的可行性，为临床此类疾病早期干预治疗探索新道路，为干细胞靶源性神经生物工程调控治疗脑脊髓损伤，促进神经纤维再生功能修复，提供新策略和实验数据。