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目的观察磷酸二酯酶Ⅳ制剂洛立普兰对髓磷脂底物上培养的大鼠大脑皮层神经元轴突生长的影响,为磷酸二酯酶Ⅳ抑制剂治疗脊髓损伤寻找依据方法预先制备髓磷脂和多聚赖氨酸底物,体外无血清培养原代大鼠皮层神经元,倒置显微镜观察神经细胞生长状态,NF200免疫组织化学染色后测量比较神经元轴突在不同底物上长度和加入洛立普兰组的长度变化,并应用RT-PCR检测GAP43mRNA的水平。结果对照组髓磷脂底物明显抑制轴突生长,且GAP-43mRNA水平降低,加入洛立普兰的神经元生长状态旺盛,克服了髓磷脂的抑制作用,突起明显增长,GAP-43mNRA的表达与空白组差异无统计学意义结论磷酸二酯酶Ⅳ抑制剂洛立普兰可以阻断髓磷脂的抑制轴突生长作用,对神经元轴突再生存在一定程度的促进作用。
OBJECTIVE To investigate the effect of loperamide, a phosphodiesterase IV preparation, on the growth of axon in rat cortical neurons cultured on myelin substrate. To find out the basis for the treatment of spinal cord injury by phosphodiesterase IV inhibitor, Polylysine substrate was used to culture primary rat cortical neurons without serum in vitro. The growth of neurons was observed under an inverted microscope. The neuronal axons were measured on different substrates by NF200 immunohistochemical staining, Plasm group length changes, and the application of RT-PCR detection of GAP43 mRNA levels. Results Myelin substrate inhibited neurite outgrowth significantly and GAP-43 mRNA level decreased. The neurons in loslitipine grew vigorously, overcame myelin inhibition, the neurite outgrowth, GAP-43 mRNA expression and the blank No significant difference was found in the group. ConclusionLupivistin, an inhibitor of phosphodiesterase IV, can block the inhibitory effect of myelin on the growth of axon and promote neuronal axon regeneration to a certain extent.