梓醇促大鼠大脑皮质神经元轴突生长的离体研究

目的：观察梓醇对原代培养的新生SD大鼠皮质神经元生存活性及轴突生长的影响。方法：新生24 h内SD大鼠大脑皮质神经元原代培养6 d后,分为空白组、终浓度分别为0.25,0.5,1.0,2.5,5.0 mg·mL-1梓醇干预组和终浓度为1.0 mg·mL^-1胞磷胆碱阳性药物对照组。药物干预48 h后,观察细胞生长情况,用NF-200免疫组化染色鉴定神经元,MTT法检测神经元生存活性,测微尺测量神经元轴突长度。结果：梓醇终浓度在1～5 mg·mL^-1时,可明显促进神经元轴突生长,2.5 mg·mL^-1时作用最强；梓醇各浓度组神经元生存活性与空白组和胞磷胆碱组差异无显著性。结论：梓醇能明显促进皮质神经元轴突生长,但对其生存活性无显著影响。

Study of catalpol promoting axonal growth for cultured cortical neurons from rats

OBJECTIVE: To explore the effects of different concentration of catalpol on the cell survival and axonal growth of cortical neurons cultured in vitro from 24 h newly born rat. METHOD: Primary cultured cortical neurons from 24 h newly born rat were dissociated and cultured. The different concentration of catalpol and 1 mg mL(-1) citicoline were added to the culture plates for 48 h, and the final of catalpol concentration were 0.25, 0.5, 1, 2.5, 5 mg mL(-1), respectively. The cortical neuron was identified by NF-200 antigen and its survival activity detected by MTT assay. The axonal growth of cultured cortical neuron were observed by inverted microscopy with micrometer. RESULT: Immunocytochemistry demonstrated more than 95% of the primary cultured cortical neurons were positive for NF-200 antigen, which indicated the cultured cells were neurons. Neurons survived growing on the concentration of 0.25, 0.5, 1, 2.5, 5 mg mL(-1). Compared with blank and 1 mg mL(-1) citicoline group,neurons survival rates were not statistical significant difference. However, it demonstrated that catalpol significantly promoted axonal growth from 1-5 mg mL(-1) (P <0.05). Interestedly, compared with the dose of 2.5 mg mL(-1), axonal growth was shorter at the dose of 5 mg mL(-1), and 2.5 mg mL(-1) catalpol showed the strongest promotion effect. CONCLUSION: The catalpol can enhance cortical neuron axonal growth, but not promote cortical neuron survival.