α-神经毒素的分离纯化及其经小鼠直肠给药后肠道体外荧光成像分析
Isolation and purification of α-neurotoxin and fluorescence imaging analysis of intestinal tract after rectum administration in mice
张昊 1廖明 2张学荣 2周怡 2孔露平 1胡少聪 2肖曼琪 2张子彦 2罗小玲3
作者信息
- 1. 广西医科大学基础医学院,广西南宁 530021
- 2. 广西医科大学生命科学研究院,广西南宁 530021
- 3. 广西医科大学基础医学院,广西南宁 530021;广西医科大学附属肿瘤医院实验研究部,广西南宁 530021
- 折叠
摘要
目的 从眼镜蛇粗毒中分离纯化短链α-神经毒素活性成分,观察荧光染料标记的α-神经毒素经小鼠直肠给药后的肠道内分布和动态过程.方法 使用CM Sepharose Fast Flow阳离子交换层析和Sephadex G-50 凝胶过滤层析分离纯化电泳纯的α-神经毒素.将4 只C57 小鼠分为对照组和实验组,对照组采用荧光染料CY7-SE标记,实验组采用荧光染料CY7-SE与α-神经毒素相互结合.使用成像仪检测CY7-SE标记的α-神经毒素在直肠给药1、3h后肠道内的荧光强度分布情况.结果 分离纯化得到电泳纯的α-神经毒素.α-神经毒素直肠给药1h后,实验组相较于对照组的小肠段和胃的荧光强度较强,盲肠、结肠段荧光强度较弱.α-神经毒素直肠给药 3h后,实验组相较于对照组的小肠、盲肠、结肠段和胃的荧光强度均较弱.结论 从眼镜蛇粗毒中分离纯化得到的α-神经毒素经直肠给药可先富集于小鼠小肠段及胃,3 h后被肠道黏膜吸收代谢.本研究为开发神经毒素的直肠递送制剂提供了实验依据.
Abstract
Objective To isolate and purify the short-chain α-neurotoxin active ingredient from the crude venom of cobra,and to observe the distribution and dynamic process of fluorescent dye-labeled α-neurotoxin in the intestinal tract after rectal administration to mice.Methods Electrophoretically pure α-neurotoxin was isolated and purified using CM Sepharose Fast Flow cation exchange chromatography and Sephadex G-50 gel filtration chromatography.Four C57 mice were divided into two groups:control and experimen-tal.In the experimental group,the fluorescent dye CY7-SE was conjugated with α-neurotoxin to each other.The fluorescence intensity distribution of CY7-SE-labeled α-neurotoxin in the intestine was detected by using an imager after 1h and 3h of rectal administration,respectively.Results Electrophoretically pure α-neurotoxin was obtained by isolation and purification.α-neurotoxin administered rectally for 1h showed stronger fluorescence intensity in the small intestine and stomach,and weaker fluorescence intensity in the cecum and colon than that in the control group.α-neurotoxin administered rectally for 3h showed weaker fluorescence intensity in the small in-testine,cecum,colon,and stomach than that in the control group.Conclusion The α-neurotoxin isolated and purified from the crude venom of cobra was first enriched in the small intestine and stomach of mice,and then absorbed and metabolized by the intestinal muco-sa after 3h.This study provides an experimental basis for the development of a neurotoxin formulation for rectal delivery.
关键词
α-神经毒素/直肠给药/荧光成像Key words
α-neurotoxin/Rectal administration/Fluorescence imaging引用本文复制引用
出版年
2024
NETL
NSTL
维普
万方数据