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

作　　者：郭雅洁 朱淼 刘延庆[1] 陶丽[1] GUO Yajie;ZHU Miao;LIU Yanqing;TAO Li(Key Laboratory of the Toric Pathogens-Based Thera peutic Approaches of Gastric Cancer,State Administration of Traditional Chinese Medicine/College of Medicine,Yangzhou University,Yangzhou 225009,China)

机构地区：[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

出　　处：《扬州大学学报（农业与生命科学版）》2024年第1期75-83,138,共10页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(81803782)。

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(neutrophil 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(80μg·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(80μg·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。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 isolating 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 DNA-NE 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H3 

分 类 号：S853.7[农业科学—临床兽医学] R285.5[农业科学—兽医学]