NCAM在成年大鼠视神经再生模型中的表达及分布与其再生作用研究

Expression and Distribution of NCAM in the Optic Nerve Regeneration Model of Adult Rats and Its Regeneration Effect

目的探讨神经细胞黏附分子(NCAM)在成年大鼠视神经再生模型中的表达、分布及其在视神经再生中的作用。方法购置健康、雄性SD大鼠32只,随机取SD大鼠8只设为空白对照组;剩余24只大鼠采用Aβ25-35寡聚体致伤SD大鼠海马CA1区,建立阿兹海默症(AD)动物模型,并构建大鼠视神经再生模型,建模成功后随机分为模型对照组(n=8只)、NSCs移植组(n=8只)及NCAM1基因修饰神经干细胞(NSCs)移植组(n=8只)。空白对照组和模型对照组注入等体积的生理盐水,NSCs移植组植入NSCs细胞,细胞数量3×105/L;NCAM1基因修饰NSCs移植组注入NCAM1基因修饰NSCs的细胞,细胞数量3×105/L,各组均完成7 d干预,比较各组学习和记忆能力、组织学检测、海马CAI区神经元形态、NCAM、ERK1/2、JNK和P38 mRNA表达。结果NCAM1基因修饰NSCs移植组潜伏期短于NSCs移植组和模型对照组,原平台象限内停留时间、穿环次数均长(多)于NSCs移植组和模型对照组,差异均有统计学意义(P<0.05);HE染色结果表明,NCAM1基因修饰NSCs移植组、NSCs移植组视神经损伤下对较轻,且NCAM1基因修饰NSCs移植组视神经损伤不明显;NCAM1基因修饰NSCs移植组NCAM、ERK1/2、JNK和P38 mRNA水平高于NSCs移植组和模型对照组,差异有统计学意义(P<0.05)。结论NCAM在成年大鼠视神经再生模型中呈低表达,经NCAM基因修饰NSCs移植用于成年大鼠视神经再生模型中可提高NCAM的表达,有助于促进视神经再生。

Objective To investigate the expression and distribution of nerve cell adhesion molecule(NCAM)in the optic nerve regeneration model of adult rats and its role in optic nerve regeneration.Methods 32 healthy and male SD rats were purchased,and 8 SD rats were randomly selected as a blank control group;the remaining 24 rats were injured by Aβ25-35 oligomers in hippocampal CA1 area of SD rats to establish Alzheimer's(AD)animal model,and build rat optic nerve regeneration model,after successful modeling,randomly divided into model control group(n=8),NSCs transplantation group(n=8)and NCAM1 gene modified neural stem cells(NSCs)transplantation Group(n=8).The blank control group and the model control group were injected with equal volume of normal saline.The NSCs transplantation group was implanted with NSCs cells,the number of cells was 3×105/L;the NCAM1 genetically modified NSCs transplantation group was injected with NCAM1 genetically modified NSCs,and the number of cells was 3×105/L.Each group completed a 7 d intervention to compare the learning and memory abilities,histological examination,neuronal morphology of the hippocampal CAI area,NCAM,ERK1/2,JNK and P38 mRNA expressions in each group.Results The latency of NCAM1 gene-modified NSCs transplantation group was shorter than that of NSCs transplantation group and model control group.The residence time and the number of looping times in the original platform quadrant were both longer(more)than those of NSCs transplantation group and model control group,the differences were statistically significant(P<0.05);HE staining results showed that:NCAM1 gene-modified NSCs transplantation group,NSCs transplantation group had less optic nerve damage,and NCAM1 gene-modified NSCs transplantation group had less obvious optic nerve injury;NCAM1 gene-modified NSCs transplantation group NCAM,ERK1/2,JNK and P38 mRNA levels were higher than NSCs transplantation group and model control group,the difference was statistically significant(P<0.05).Conclusion NCAM has a low expression in the model of optic nerve regeneration in adult rats.Transplantation of NSCs modified with NCAM gene for the model of optic nerve regeneration in adult rats can increase the expression of NCAM and help promote the regeneration of optic nerve.