眼镜蛇神经毒素的规模化制备及其镇痛作用的研究

目的 建立一种快速、规模化制备眼镜蛇毒神经毒素的方法.方法 采用CM-sephadex C-25和CM-sepharose fast flow离子交换柱层析纯化经初步处理的眼镜蛇毒;SDS-PAGE和HPLC检测产物的纯度;离体蛙心和红细胞溶血试验检查产物的心脏毒性和溶血性;小白鼠热板模型、醋酸扭体模型和大白鼠电刺激嘶叫模型评价产物的镇痛作用.结果 制备的产物可达到电泳纯和HPLC单一峰,无心脏毒性和溶血活性,镇痛作用强,但起效慢.结论 所用工艺适合眼镜蛇毒神经毒素工业化制备的需要.

Preparation of neurotoxin from cobra venom on a large scale and its analgesic effect

OBJECTIVE To establish a method to prepare the cobra neurotoxin(CNT) on a large scale.METHODS The cobra venom was isolated and purified by successive CM-sephadex C-25 and CM-sepharose fast flow ion exchange chromatography.The purity of the product was determined by SDS-PAGE and HPLC.The cardiotoxicity and hemolytic activity of the product were investigated via the frog heart perfusion by Straub method and the hemolysis experiment,respectively.The analgesic activity of the product was also estimated by the models of hot-plate test,writhing response in mice and electrical stimulation-vocalization in rats.RESULTS The cobra neurotoxin obtained was proved to be electrophrotic pure and produced only one peak in HPLC examination.Neither cardiotoxicity nor hemolytic activity existed.Its analgesic effect on the experimental animals was strong,but with a slow onset time.CONCLUSION This method can meet the need of industrial preparation of cobra neurotoxin.