目的研究维生素C(Vitamin C,Vit.C)对大鼠背根神经节神经元轴突生长的影响,为探讨神经再生新策略提供基础。方法取新生SD大鼠背根神经节(dorsal root ganglion,DRG)进行神经元分离培养。在培养液中加入不同浓度的维生素C(0,100,200,400μmol/L)处理24 h后通过βIII tubulin,Rho A免疫组化以及鬼笔环肽染色对神经元突起的数量和长度、生长锥伪足数量、神经元胞体大小及其内Rho A表达强度进行分析。结果神经元突起的数量、长度,生长锥伪足的数量以及神经元胞体面积均随维生素C的浓度增高而增长,而神经元内Rho A表达强度则相应降低。结论维生素C可促进体外大鼠背根节神经元突起的生长,这可能与Rho A表达下调相关。
Objective To study the effect of Vitamin C on the neurite outgrowth of dorsal root ganglionic neurons. Methods Neurons harvested from the dorsal root ganglion of newborn SD rat pups were cultured in a basic medium supplemented with a series of concentration of Vitamin C (0, 100, 200, 400μmol/L). 24 hours later, the cultures were immunostained with antibodies of βⅢ tubulin and RhoA. The βⅢ tubulin immunostained cultures were double stained with phalloidin. Then, the number and length of the neurites, number of pseudopodium in the growth cone of the cultured neurons, as well as the immunofluorence intensity of RhoA in the neuronal bodies were quantified. Results The number and length of the neurites, number of pseudopodium in the growth cone of the cultured neurons were all increased in Vit. C treated groups compared with the control group. Howerver, the RhoA level was decreased in the Vit. C treated groups. Conclusion Vit.C could promote the neurite outgrowth of cultured dorsal root ganglionic neurons which may result in reducing the expression of RhoA.