基于原代小脑颗粒细胞的A型肉毒毒素活性分析法的建立

Evaluation of activity of botulinum neurotoxin A based on primary cerebellar granulosa cells

目的分离、培养SD大鼠原代小脑颗粒细胞,用于评价A型肉毒毒素活性。方法将出生后6~8 d的SD大鼠小脑颗粒细胞培养5~7 d后用A型肉毒毒素处理,采用免疫荧光法进行检测并评价毒素的活性,Western印迹法分析毒素活性与剂量关系。结果与结论成功培养了大鼠小脑颗粒细胞,并在原代神经细胞水平建立了A型肉毒毒素活性分析法,测定了毒素活性剂量关系,为进一步解析肉毒毒素作用的生化机制提供了技术手段。

Objective To isolate and culture the primary cerebellar granulosa cells （CGNs） of SD rats and evaluate the activity of botulinum ueurotoxin A （BoNT/A） based on CGNs. Methods CGNs of 6-to 8-days-old SD rats were isolated and cultured. After 5 - 7 d, the cells were treated with BoNT/A. The activity of the toxin was evaluated with immunofluo- rescence, and the relationships between the activity and dose of the toxin were analyzed with Western blotting. Results and Conclusion CGNs Of SD rats were successfully cultured, a method for evaluating the activity of BoNT/A was established at the level of primary nerve cells, and the relationships between the activity and dose of toxin were analyzed. This study provides a tool for further detailing the biochemical mechanism of BoNT/A.