活细胞成像技术研究2,5-己二酮对中分子量神经丝运输的影响

Using live-cell imaging technique to study the effect of 2,5-HD on axonal transport of medium-molecular neurofilament

目的用活细胞成像技术研究2,5-己二酮对中分子量神经丝蛋白(NF-M)在体外培养的背根神经节神经元轴突中的运输的影响。方法转染pPA-GFP-NFM质粒的背根神经节神经元体外培养,给予2,5-己二酮作用24 h后,荧光显微镜下定量观察GFP-NFM的转运量。结果给药4、8和16 mmol/L 2,5-HD组,NF-M在轴突中被转运的量分别为28.5±11.6%(P<0.05)、22.6±9.2%(P<0.01)和18.5±8.6%(P<0.01),而空白对照组中NF-M被转运的量为35.6±11.0%。结论 2,5-己二酮降低了NF-M的轴浆运输速率,这可能是2,5-己二酮引起中毒性周围神经病的致病机制之一。

Objective To explore the effects of 2,5-Hexanedione（2,5-HD）-induced axonal transport of NF-M in rat DRG cells. Methods The cultured DRG neurons were transfected with pPA-GFP-NFM plasmid. After the cells were treated with 0, 4, 8, 16 mmol/L 2,5-HD for 24 hours, the technique of live-cell imaging was performed, and the fluorescence images were collected dynamically at each 30 minute intervals. The amount of NF-M movements was detected by the decrease of the fluorescence intensity. Results The results showed that 2,5-HD slowed the NF-M movement in axons of cultured DRG cells. The pulse-escape experiments of 24 h showed that the NF-M were escaped by 28.5 ± 11.6% （P 〈0.05）, 22. 6 ±9. 2% （P 〈0. 01） and 18.5 ±8.6% （P 〈0.01） in 4, 8, 16 mmol/L 2,5-HD groups when compared to the initial value （ 100% ） ; while that was decreased by 35.6 ± 11.0% in control cells. Conclusion 2,5-Hexanedione （2,5- HD） could slow the NF-M transport in axons of rat DRG neurons, and that may contribute to n-hexane-induced neurotoxicity.