目的 1研究人类脐血干细胞在治疗大鼠外伤性视神经损伤的有效性;2 初步探讨人类脐血干细胞在治疗外伤性视神经损伤过程中的作用机制;3 为人类脐血干细胞应用于视神经损伤病人提供理论依据.
方法 1 建立致伤强度量化的大鼠外伤性视神经损伤模型;2 采用随机对照的方法,以尾静脉注射,球旁注射及玻璃体腔注射三种方法将人类脐血干细胞,神经营养因子,以及2者的混合物分别移植入外伤性视神经损伤大鼠体内,对照组采用脐血干细胞载体,观察各组疗效.3采用随机对照的方法,以尾静脉注射,球旁注射及玻璃体腔注射三种方法将荧光标记的人类脐血干细胞移植入外伤性视神经损伤大鼠体内,通过激光扫描共聚焦显微镜初步观察干细胞的迁移,分布;通过视网膜铺片,视网膜石蜡切片,视网膜受损处石蜡切片于荧光显微镜下观察干细胞的分布和分化;用RT-PCR半定量测定视网膜内脑源性神经营养因子(BDNF),胶质细胞源神经营养因子(GDNF)和睫状神经营养因子(CDNF)MRNA的相对含量.并检测脐血干细胞移植对于外伤性视神经损伤后视网膜神经节细胞凋亡的影响。
结果 与对照组相比,脐血干细胞移植后视网膜内神经营养因子(BDNF,GDNF,CDNF)均持续表达(P<0.05),GRP78(葡萄糖调节蛋白78)及CHOP(CCAAT/增强子结合蛋白同源蛋白)均降低(P<0.05),减轻了内质网应激的强度,减少了损伤缺血区域的视网膜神经节细胞的凋亡并防止残存的神经节细胞凋亡。
结论 脐血干细胞能在视神经损伤的大鼠体内存活,迁移,分化,并能改善其神经功能;此外,脐血干细胞还有明显的抗凋亡作用.
Objective 1 To study the effectiveness of human umbilical cord blood stem cells in the treatment of traumatic optic nerve damage in rats; 2 TO explore the mechanism of human umbilical cord blood stem cells in the treatment of traumatic optic nerve injury ; 3 In order to provide a theoretical basis for the The clinical application of human umbilical cord blood stem cells.
Methods Establish the rat model of traumatic optic nerve injury which is quantified; 2 By the randomized controlled way , Implanted the human umbilical cord blood stem cells, neurotrophic factors and the mixture into the traumatic optic nerve injury rats by three injection methods of tail vein, vitreous cavity and periglomerular. Observe effect of each group including the control one with umbilical cord blood stem cell carrier.By the randomized controlled way , Implanted the human umbilical cord blood stem cells by fluorescent labelling into the traumatic optic nerve injury rats by three injection methods of tail vein, vitreous cavityand periglomerular. Observe the migration and distribution of the stem cell by laser scanning confocal microscope. Observe the distribution and differentiation of the stem cell Through the fluorescence microscope by the stretched preparation, paraffin section of retina and the paraffin section of impaired retina.Determin the relative amount (semi-determinalion) of BDNF, GDNF and CDNF mRNA in retina by RT-PCR.And determine the effect of RCGs apoptosis after traumatic optic nerve injury ,which Is induced by the umbilical cord blood stem cells explantation.
Results Compared with the control group,neurotrophic factors (BDNF, GDNF,CDNF) have continued to express after umbilical cord blood stem cell transplantation , (P<0.05). GRP78 and CHOP are also reduced(p<0.05)which lead to Mitigate the intensity of endoplasmic reticulum stress and RCGs opoptosis ,furthermore it can Strengthen The prevention of apoptosis of Residual RCGs cells.
Conclusion The umbilical cord blood stem cells can not only survive, migration, differentiation in rats of traumatic optic nerve injury but also improve nerve function .In addition, the umbilical cord blood stem cells have significant anti-apoptosis effect. |