A型肉毒素重链干预神经细胞组蛋白3乙酰化水平的初步研究  

Botulinum neurotoxin serotype A heavy chain intervenes in the H3 acetylation: A preliminary study

作　　者：兰婧[1] 刘福[1] 高扬 李志强[2] 刘雅 赵明伟 袁海霞 李夏青[1] LAN Jing;LIU Fu;GAO Yang;LI Zhiqiang;LIU Ya;ZHAO Mingwei;YUAN Haixia;LI Xiaqing(Department of Pathophysiology,Taiyuan 030001,China;The First Clinical Medical College,Shanxi MedicalUniversity,Taiyuan 030001)

机构地区：[1]山西医科大学病理生理教研室,太原030001 [2]山西医科大学第一临床医学院,太原030001

出　　处：《中国比较医学杂志》2018年第6期4-9,共6页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81171178);山西医科大学大学生创新创业基金(20170105)

摘　　要：目的通过对离体神经细胞培养及大鼠在体脊髓损伤模型施予BoNT/A重链,观察损伤局部组蛋白乙酰化水平的变化,为探讨BoNT/A重链干预神经细胞再生的分子机制提供实验依据。方法将BoNT/A重链加入细胞培养液或在脊髓损伤模型基础上局部给予小剂量BoNT/A重链;收集细胞及损伤局部脊髓组织,采用免疫荧光、SDS-PAGE和Western blot等方法对细胞和脊髓组织内选择性组蛋白亚类的乙酰化水平进行检测。结果不论是体内应用还是培养液内加入BoNT/A重链,细胞和组织内组蛋白3的乙酰化水平皆明显增多;BoNT/A重链所致的Neuro-2a细胞组蛋白3乙酰化水平的增高与神经突起的增长相匹配,即同一时间点组蛋白乙酰化水平显著增高的同时,神经突起的生长也呈显著增长;脊髓损伤基础上给予BoNT/A重链所显示的组蛋白3乙酰化的表达量增高呈现双时相高峰(2 h和48 h)。结论 BoNT/A重链可促进组蛋白3的乙酰化;在同一时间点,组蛋白3的乙酰化水平与神经突起增长的长度相一致;组蛋白3乙酰化可能是BoNT/A重链促神经突起生长的相关机制之一。Objective To investigate the effect and molecular mechanism of botulinum neurotoxin serotype A（BoNT/ A） heavy chain on neuron regeneration. Methods Cell culture, rats, immunofluorescence, SDS-PAGE andwestern blot, etc. were adopted in this study to explore the alterations of histone-3 acetylation （ acetyl-H3 ） by localtreatment of BoNT/ A heavy chain to spinal cord injury （SCI） in rats （in vivo） or by adding it into cell culture （in vitro）.Meanwhile, the relevance of acetyl-H3 to neurite out-growth based on SCI and cell culture with BoNT/ A heavy chainapplication was approached as well. Results The application of BoNT/ A heavy chain to cultured Neuro-2a cells increasedthe level of H3 acetylation. The increase of H3 acetylation was paralleled with the growth of neuritogenesis. Also, theneuronal treatment of BoNT/ A heavy chain to SCI promoted the re-growth of neuronal processes surrounding the lesions.The growth of neuronal processes was positively correlated to the level of H3 acetylation. During the periods of BoNT/ A heavy chain treatment in vivo or in vitro, the increase of H3 acetylation showed two peaks. Conclusions BoNT/ A heavychain increased the H3 acetylation, which might be one of its neuritogenic mechanisms.

关 键 词：A型肉毒素 A型肉毒素重链 组蛋白乙酰化 乙酰化/去乙酰化 神经突起再生 脊髓损伤 

分 类 号：R-33[医药卫生]