Wan Dong, Zhu Hui-feng, Luo Yong, Xie Peng, Xu Xiao-yu
Department of Neurology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Zhongguo Zhong Yao Za Zhi. 2007 Sep;32(17):1771-4.
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.
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.
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.
The catalpol can enhance cortical neuron axonal growth, but not promote cortical neuron survival.
探讨不同浓度梓醇对新生24 h大鼠体外培养皮质神经元细胞存活及轴突生长的影响。
将新生24 h大鼠的皮质神经元进行原代培养、分离。在培养板中加入不同浓度的梓醇及1 mg/mL胞磷胆碱,作用48 h,梓醇终浓度分别为0.25、0.5、1、2.5、5 mg/mL。采用NF-200抗原鉴定皮质神经元,MTT法检测其存活活性。用带测微尺的倒置显微镜观察培养的皮质神经元轴突生长情况。
免疫细胞化学显示,原代培养的皮质神经元中NF-200抗原阳性率>95%,表明所培养细胞为神经元。神经元在0.25、0.5、1、2.5、5 mg/mL浓度下均能存活生长。与空白组和1 mg/mL胞磷胆碱组比较,神经元存活率差异无统计学意义。但结果显示,梓醇在1~5 mg/mL能显著促进轴突生长(P<0.05)。有趣的是,与2.5 mg/mL剂量组比较,5 mg/mL剂量组轴突生长较短,2.5 mg/mL梓醇促进作用最强。
梓醇可促进皮质神经元轴突生长,但对皮质神经元存活无促进作用。