南蛇藤总萜抑制中性粒细胞胞外诱捕网形成的作用与机制研究
The effect and mechanism of the total terpenoids of Celastrus orbiculatus Thunb.on inhibiting the formation of neutrophil extracellular traps
郭雅洁 1朱淼 1刘延庆 1陶丽1
作者信息
- 1. 扬州大学医学院/国家中医药管理局胃癌毒邪重点论治研究室,江苏扬州 225009
- 折叠
摘要
为探索南蛇藤总萜(the total terpenoids of Celastrus orbiculatus Thunb.,TTC)对中性粒细胞胞外诱捕网(neu-trophil extracellular traps,NETs)形成及其作用机制,通过分离与制备中性粒细胞,建立佛波酯(phorbol 12-myristate 13-acetate,PMA)诱导的NETs形成细胞模型.利用瑞氏-姬姆萨染色、扫描电镜与免疫荧光技术研究南蛇藤总萜对NETs形成及其生物学标志的影响,并利用Annexin V/PI试验研究南蛇藤总萜对中性粒细胞凋亡的影响.结果表明:南蛇藤总萜能剂量依赖性地抑制PMA诱导的NETs形成;南蛇藤总萜(80 μg·mL-1)可阻断中性粒细胞弹性蛋白酶(neutrophil elastase,NE)的释放,并破坏NETs相关DNA-NE复合物的形成;南蛇藤总萜(80 μg·mL-1)可显著抑制PMA诱导的瓜氨酸化组蛋白H3(citrullinated histone H3,CitH3)的表达水平;南蛇藤总萜可剂量依赖性地促进中性粒细胞凋亡.综上,南蛇藤总萜对中性粒细胞NETs的形成具有抑制作用,其机制可能与其直接诱导中性粒细胞凋亡有关.
Abstract
To investigate the mechanisms and the effect of the total terpenoids of Celastrus orbiculatus Thunb.(TTC)on NETs,the cell model of NETs formation induced by phorbol 12-myristate 13-acetate(PMA)was established by isola-ting and preparing neutrophils.The effects of TTC on the formation of NETs and their biological markers were studied by Wright-Giemsa Staining,scanning electron microscope and immunofluorescence.The Annexin V/PI experiment was used to study TTC on neutrophil apoptosis.Results showed that TTC could dose-dependently inhibit the formation of NETs induced by PMA;TTC(80 μg·mL-1)could block the release of neutrophil elastase(NE)and disrupt the formation of Nets associated DNANE complexes.TTC(80 μg·mL-1)significantly inhibited PMA-induced CitH3 expression;TTC can promote neutrophil apoptosis in a dose-dependent manner.This study suggests that TTC can inhibit the formation of neutrophil NETs,and its mechanism may be related to its direct induction of neutrophil apoptosis.
关键词
南蛇藤总萜/中性粒细胞胞外诱捕网/凋亡/中性粒细胞弹性蛋白酶/瓜氨酸化组蛋白H3Key words
Celastrus orbiculatus Thunb./neutrophil extracellular traps/apoptosis/neutrophil elastase/citrullinated histone H3引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据