胎肝干细胞联合人参皂苷对大鼠脊髓损伤的实验研究
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摘要
脊髓损伤是中枢神经系统的一种严重创伤，虽然手术治疗和药物治疗在不同程度上使受损的脊髓得到了恢复和再生的空间，但仍不可避免的造成了病人瘫痪乃至死亡的严重后果。脊髓损伤经常是在意外发生时，由于原发性机械损害及继发性损伤，导致组织中部分或者全部的神经细胞、神经胶质细胞等出现死亡现象，同时组织中的神经传导纤维发生断裂、脱髓鞘等，从而造成了脊髓神经功能的缺失。由于神经元的独特性，即神经元具有不可再生性，加之在脊髓发生损伤后，局部脊髓组织内复杂的微环境的改变，从而抑制了脊髓组织中神经元的轴突再生，这些病理改变最终使得脊髓中死亡的神经元功能丧失，导致器官和系统失神经支配，给患者及家人造成了极大的痛苦和生活上的不便。
人参中含有多种有效成分，包括丰富的人参皂苷、多种氨基酸、多糖及挥发油等成分，对人体的中枢神经系统、心血管系统、免疫系统及泌尿生殖系统等具有广泛的作用，能增强机体对有害刺激的抵抗力。研究者在研究人参功效时，发现人参皂苷、三萜类物质，尤其是人参皂苷对受损脊髓具有良好的修复作用。
近年来，随着干细胞研究的不断深入，研究者尝试利用胚胎干细胞修复损伤的脊髓组织，并且成为目前的一个研究热点。干细胞移植治疗神经再生的可移植性，为脊髓损伤的治疗带来了光明。将诱导后的神经细胞移植入脊髓中，可促使髓鞘再生，促使宿主神经元间再次形成突触。研究认为是新植入的神经细胞整合到宿主神经细胞中，因而部分恢复了脊髓信号传导的能力。
众所周知，干细胞是一类全能的细胞，也是指一类具有自我更新、复制能力，并且具有多种分化潜能的细胞，他们可以在适宜条件下分化成特定类型的细胞，应用他们可以替代已经丧失功能、失去修复能力的细胞，从而达到修复组织功能的功效。
研究发现，具有修复能力的干细胞包括胚胎干细胞（ESC）、骨髓间充质干细胞（MSC）、神经干细胞（NSC）等。而用于中枢神经系统修复的干细胞主要为胚胎干细胞和神经干细胞。目前少有利用胎肝干细胞诱导成神经干细胞进而完成神经系统修复的研究，且胎肝干细胞尚缺乏特异性的检测标志物。本实验分离的肝间充质干细胞表达CD44、CD29、CD105，说明分离的肝间充质干细胞具有低免疫原性，有利于异基因的细胞移植。对其研究，有可能发挥其稳定性好等优点，从而有可能成为应用于临床治疗脊髓损伤等中枢神经系统损伤的重要手段之一。
本实验证实了胎肝间充质干细胞具有向神经细胞分化的潜能。诱导后细胞具有典型神经细胞形态，同时表达神经细胞相关蛋白。nestin和NSE在诱导后细胞中表达，GFAP不表达，证实肝间充质干细胞可以向神经元样细胞分化，而不分化为神经胶质细胞。进而利用其与人参皂苷共同治疗脊髓损伤的大鼠，发现其可以明显改善大鼠脊髓损伤症状，恢复其运动功能。
本课题利用Al1en’s理论构建大鼠脊髓损伤模型作为研究脊髓损伤修复的研究模型，同时证明胎肝间充质干细胞可以诱导分化为神经元样细胞，进而辅助以人参皂苷治疗脊髓损伤，旨在探讨二者联合可以通过降低脊髓受损组织细胞中的活性氧水平，促进细胞抗凋亡，来降低脊髓损伤的程度，促进其功能恢复，这为临床脊髓损伤修复提供了重要的参考依据，具有重要的指导意义，使其有望成为组织工程和细胞移植治疗的新技术。
Spinal cord injury is a serious central nervous system trauma, surgical treatmentand medication to ease the change of spinal cord injury in varying degrees, but theconsequences still inevitably result in a patient with paraplegia. Spinal cord injury isoften occured at the time of the accident, as a result of the primary mechanicaldamage and secondary injury, resulting organization part or all of the nerve cells, glialcells died, and nerve conduction in the tissue fibers broken. The demyelinatingresulting lack of function of the spinal cord. Due to the uniqueness of the neurons,neurons with non-renewable, and spinal cord injury after partial spinal cord tissuemicroenvironment change, thereby inhibiting axonal regeneration in spinal cord tissue,ultimately makes the spinal cord death loss of neuronal function, causedinconvenience and pain of life for patients and their families.
Contain a variety of active ingredients, including a wealth of ginsenoside inginseng, a variety of amino acids, polysaccharides and volatile oil components, andhas an extensive role in the body's central nervous system, cardiovascular system,immune system and urogenital system, can enhance the body The noxious stimuliresistance. The researchers found ginsenoside of triterpenoids ginseng efficacy study,ginsenoside has a good role in the repair of spinal cord injury.
In recent years, with the development of the stem cell research, the researchersare trying to use of embryonic stem cells to repair damaged spinal cord tissue, and thisresearch are becoming a research focus. Stem cell transplantation in the treatment ofnerve regeneration portability, brought to light for the treatment of spinal cord injury.After the induction of nerve cells transplanted into the spinal cord, can promoteremyelination, prompting host neurons form synapses again. The study suggests thatthe new implantable nerve cells integrated into the host nerve cells, and thus the partial restoration of the ability of the spinal cord to transmit signals.
As all we known, the stem cells are stem cells is a Class wholly competent cells,capable differentiation as a specific cell type, also refers to a class having aself-renewal and the ability to replicate, and having a variety of differentiationpotential of the cells, they can be under suitable conditions differentiation into aparticular type of cells, the applications that they can replace the loss of function, lossof the ability to repair cells, so as to achieve the efficacy of the repair tissue function.
Our study found that, with the repair capacity of stem cells, including embryonicstem cells (ESC), bone marrow mesenchymal stem cells (MSC), neural stem cells(NSC). Stem cells for repair of the central nervous system of embryonic stem cellsand neural stem cells. Currently little data on personnel with fetal liver stem cells intoneural stem cells and then to complete the study of the nervous system repair. Atpresent, no specific sign of liver stem cells have not. The experimental separation ofMSCs expressed CD44, CD29, CD105, separation liver MSCs cells has lowimmunogenicity favor the allogeneic cell transplantation. Research, are likely to playa good stability, which is likely to become one of the important means used in theclinical treatment of spinal cord injury and other central nervous system damage.
The present study demonstrated that liver MSCs have the potential todifferentiate into nerve cells. Typical nerve cell morphology induced cells, whileexpression of the related proteins of nerve cells. nestin and NSE in the cells afterinduction of expression, GFAP expression, confirmed liver MSCs can differentiateinto neuron-like cells, without having to differentiate into glial cells. Then use itsginsenoside treatment of spinal cord injury in rats, found that spinal cord injury cansignificantly improve symptoms, restore their motor function.
The theory of the the subjects of Al1en the to build rat model of spinal cordinjury as a research model of spinal cord injury repair, at the same time prove fetalliver stem cells can be induced to differentiate into neuron-like cells, and further helpto the treatment of spinal cord injury ginsenoside aims to explore both importantreference can be provided jointly by reducing ROS levels in the spinal cord damaged tissue cells, promoting cell resistance to apoptosis, and to reduce the damage of thespinal cord injury, and promote functional recovery, clinical spinal cord injury repairhas important guiding significance, it is expected to become the new technologies fortissue engineering and cell transplantation in the treatment.
人参中含有多种有效成分，包括丰富的人参皂苷、多种氨基酸、多糖及挥发油等成分，对人体的中枢神经系统、心血管系统、免疫系统及泌尿生殖系统等具有广泛的作用，能增强机体对有害刺激的抵抗力。研究者在研究人参功效时，发现人参皂苷、三萜类物质，尤其是人参皂苷对受损脊髓具有良好的修复作用。
近年来，随着干细胞研究的不断深入，研究者尝试利用胚胎干细胞修复损伤的脊髓组织，并且成为目前的一个研究热点。干细胞移植治疗神经再生的可移植性，为脊髓损伤的治疗带来了光明。将诱导后的神经细胞移植入脊髓中，可促使髓鞘再生，促使宿主神经元间再次形成突触。研究认为是新植入的神经细胞整合到宿主神经细胞中，因而部分恢复了脊髓信号传导的能力。
众所周知，干细胞是一类全能的细胞，也是指一类具有自我更新、复制能力，并且具有多种分化潜能的细胞，他们可以在适宜条件下分化成特定类型的细胞，应用他们可以替代已经丧失功能、失去修复能力的细胞，从而达到修复组织功能的功效。
研究发现，具有修复能力的干细胞包括胚胎干细胞（ESC）、骨髓间充质干细胞（MSC）、神经干细胞（NSC）等。而用于中枢神经系统修复的干细胞主要为胚胎干细胞和神经干细胞。目前少有利用胎肝干细胞诱导成神经干细胞进而完成神经系统修复的研究，且胎肝干细胞尚缺乏特异性的检测标志物。本实验分离的肝间充质干细胞表达CD44、CD29、CD105，说明分离的肝间充质干细胞具有低免疫原性，有利于异基因的细胞移植。对其研究，有可能发挥其稳定性好等优点，从而有可能成为应用于临床治疗脊髓损伤等中枢神经系统损伤的重要手段之一。
本实验证实了胎肝间充质干细胞具有向神经细胞分化的潜能。诱导后细胞具有典型神经细胞形态，同时表达神经细胞相关蛋白。nestin和NSE在诱导后细胞中表达，GFAP不表达，证实肝间充质干细胞可以向神经元样细胞分化，而不分化为神经胶质细胞。进而利用其与人参皂苷共同治疗脊髓损伤的大鼠，发现其可以明显改善大鼠脊髓损伤症状，恢复其运动功能。
本课题利用Al1en’s理论构建大鼠脊髓损伤模型作为研究脊髓损伤修复的研究模型，同时证明胎肝间充质干细胞可以诱导分化为神经元样细胞，进而辅助以人参皂苷治疗脊髓损伤，旨在探讨二者联合可以通过降低脊髓受损组织细胞中的活性氧水平，促进细胞抗凋亡，来降低脊髓损伤的程度，促进其功能恢复，这为临床脊髓损伤修复提供了重要的参考依据，具有重要的指导意义，使其有望成为组织工程和细胞移植治疗的新技术。
Spinal cord injury is a serious central nervous system trauma, surgical treatmentand medication to ease the change of spinal cord injury in varying degrees, but theconsequences still inevitably result in a patient with paraplegia. Spinal cord injury isoften occured at the time of the accident, as a result of the primary mechanicaldamage and secondary injury, resulting organization part or all of the nerve cells, glialcells died, and nerve conduction in the tissue fibers broken. The demyelinatingresulting lack of function of the spinal cord. Due to the uniqueness of the neurons,neurons with non-renewable, and spinal cord injury after partial spinal cord tissuemicroenvironment change, thereby inhibiting axonal regeneration in spinal cord tissue,ultimately makes the spinal cord death loss of neuronal function, causedinconvenience and pain of life for patients and their families.
Contain a variety of active ingredients, including a wealth of ginsenoside inginseng, a variety of amino acids, polysaccharides and volatile oil components, andhas an extensive role in the body's central nervous system, cardiovascular system,immune system and urogenital system, can enhance the body The noxious stimuliresistance. The researchers found ginsenoside of triterpenoids ginseng efficacy study,ginsenoside has a good role in the repair of spinal cord injury.
In recent years, with the development of the stem cell research, the researchersare trying to use of embryonic stem cells to repair damaged spinal cord tissue, and thisresearch are becoming a research focus. Stem cell transplantation in the treatment ofnerve regeneration portability, brought to light for the treatment of spinal cord injury.After the induction of nerve cells transplanted into the spinal cord, can promoteremyelination, prompting host neurons form synapses again. The study suggests thatthe new implantable nerve cells integrated into the host nerve cells, and thus the partial restoration of the ability of the spinal cord to transmit signals.
As all we known, the stem cells are stem cells is a Class wholly competent cells,capable differentiation as a specific cell type, also refers to a class having aself-renewal and the ability to replicate, and having a variety of differentiationpotential of the cells, they can be under suitable conditions differentiation into aparticular type of cells, the applications that they can replace the loss of function, lossof the ability to repair cells, so as to achieve the efficacy of the repair tissue function.
Our study found that, with the repair capacity of stem cells, including embryonicstem cells (ESC), bone marrow mesenchymal stem cells (MSC), neural stem cells(NSC). Stem cells for repair of the central nervous system of embryonic stem cellsand neural stem cells. Currently little data on personnel with fetal liver stem cells intoneural stem cells and then to complete the study of the nervous system repair. Atpresent, no specific sign of liver stem cells have not. The experimental separation ofMSCs expressed CD44, CD29, CD105, separation liver MSCs cells has lowimmunogenicity favor the allogeneic cell transplantation. Research, are likely to playa good stability, which is likely to become one of the important means used in theclinical treatment of spinal cord injury and other central nervous system damage.
The present study demonstrated that liver MSCs have the potential todifferentiate into nerve cells. Typical nerve cell morphology induced cells, whileexpression of the related proteins of nerve cells. nestin and NSE in the cells afterinduction of expression, GFAP expression, confirmed liver MSCs can differentiateinto neuron-like cells, without having to differentiate into glial cells. Then use itsginsenoside treatment of spinal cord injury in rats, found that spinal cord injury cansignificantly improve symptoms, restore their motor function.
The theory of the the subjects of Al1en the to build rat model of spinal cordinjury as a research model of spinal cord injury repair, at the same time prove fetalliver stem cells can be induced to differentiate into neuron-like cells, and further helpto the treatment of spinal cord injury ginsenoside aims to explore both importantreference can be provided jointly by reducing ROS levels in the spinal cord damaged tissue cells, promoting cell resistance to apoptosis, and to reduce the damage of thespinal cord injury, and promote functional recovery, clinical spinal cord injury repairhas important guiding significance, it is expected to become the new technologies fortissue engineering and cell transplantation in the treatment.
引文
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