脑源性神经营养因子通过PI3K/Akt信号通路促进大鼠坐骨神经再生修复的研究  

Brain-derived neurotrophic factor promotes regeneration and repair of rat sciatic nerve through PI3K/Akt signaling pathway

作　　者：李想 毕佳璇 孙秋霞 于润泽 贾斯祺 付秀美 LI Xiang;BI Jiaxuan;SUN Qiuxia;YU Runze;JIA Siqi;FU Xiumei(Department of Human Anatomy,School of Basic Medicine,Chengde Medical University,Chengde 067000,China;Hebei Key Laboratory of Nerve Injury and Repair,Chengde 067000,China)

机构地区：[1]承德医学院基础医学院人体解剖学教研室,河北承德067000 [2]河北省神经损伤与修复重点实验室,河北承德067000

出　　处：《中国医科大学学报》2023年第1期51-56,共6页Journal of China Medical University

基　　金：河北省自然科学基金(H2021406056);河北省高等学校科学研究计划(ZD2020178);国家级大学生创新创业训练计划(2020003);河北省科技厅“技术创新引导专项-科技工作会商”项目(2020)。

摘　　要：目的 探讨脑源性神经营养因子(BDNF)促进坐骨神经再生修复的作用及其机制。方法 选取10只SPF级雄性SD大鼠制备脱细胞神经移植物(ANA)。取5只大鼠设为正常(N)组。另取20只大鼠建立坐骨神经损伤模型,然后再随机分为模型(M)组、ANA组、LY294002(LY)组、溶剂对照(LYC)组。M组不再处理,其余3组均桥接ANA于损伤神经的两断端处,LY组和LYC组分别于桥接术后第2日腹腔注射LY294002或DMSO(2.0 ng·mL^(-1)·kg^(-1)),连续注射4周。测定各组大鼠运动神经传导速度(MNCV)和胫前肌湿重比率,尼氏染色观察各组大鼠脊髓前角运动神经元的形态结构,免疫荧光染色和Western blotting法检测脊髓内BDNF和p-Akt1蛋白的表达。结果 与M组比较,ANA组大鼠MNCV和胫前肌湿重比率明显升高(P <0.05);与ANA组比较,LY组MNCV和胫前肌湿重比率显著降低(P <0.05)。尼氏染色结果显示,各组脊髓内均可见蓝紫色呈斑块状的尼氏体,定量分析结果显示ANA组尼氏体数量明显多于M组(P <0.05),LY组尼氏体数量少于ANA组(P <0.05)。免疫荧光染色和Western blotting结果显示,ANA组脊髓内BDNF和p-Akt1蛋白表达量均高于M组(P <0.05);LY组脊髓BDNF和p-Akt1蛋白表达量显著低于ANA组(P <0.05)。结论 BDNF可通过激活PI3K/Akt信号通路发挥促进大鼠神经再生修复的作用。Objective To investigate the effect and mechanism of brain-derived neurotrophic factor(BDNF) on sciatic nerve regeneration and repair. Methods Ten male Sprague Dawley(SD) rats were randomly selected to prepare the acellular nerve graft(ANA).The normal(N) group comprised 5 male SD rats. The sciatic nerve injury model was established in 20 male SD rats;then,the model rats were randomly divided into the model(M),ANA,LY294002(LY),and solvent control(LYC) groups. The rats in the M group received no further treatment. Considering the other three groups,the rats were bridged with the ANA;two days after ANA bridging,rats in the LY and LYC groups were administered LY294002 and dimethyl sulfoxide,respectively,at 2.0 ng·ml^(-1)·kg-1,for a total of four weeks. For each group,the motor nerve conduction velocity(MNCV) and the wet weight ratio of the tibialis anterior muscles were determined. Nissl stating was performed to examine the morphology of motor neurons of the anterior horn in the spinal cord. Expression of BDNF and p-Akt1was determined by immunofluorescence and Western blotting. Results Compared with the M group,the ANA group presented elevated values of MNCV and the wet weight ratio of tibialis anterior muscles(P < 0.05). Compared with the ANA group,the MNCV and the wet weight ratio of tibialis anterior muscles were reduced in the LY group(P < 0.05). Nissl staining revealed blue-purple plaque Nissl bodies in each group. Based on quantitative analysis,the ANA group exhibited more Nissl bodies than the M group(P < 0.05),whereas the LY group showed fewer Nissl bodies than the ANA group(P < 0.05). Expression of BDNF and p-Akt1 in the ANA group was relatively higher than that in the M group(P < 0.05);the expression was relatively lower in the LY group than that in the ANA group(P < 0.05).Conclusion BDNF could promote nerve regeneration and repair by activating the PI3K/Akt pathway.

关 键 词：BDNF PI3K/AKT信号通路 脊髓 坐骨神经损伤 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]