Trimetazidine reduces formation of neutrophil extracellular traps via inhibiting autophagy / 中国药理学通报

ABSTRACT

Aim To study the effects of trimetazidine (TMZ)on theformationof neutrophil extracellular traps (NETs) induced by oxidized low-density lipoprotein (ox-LDL) in vitro and its relationship associated with cell autophagy. Methods The bone marrow neutrophils of mice were extracted by density gradient centrifugation,and NETs induction model was established using ox-LDL. Furthermore, TMZ, autophagy inhibitor LY294002 and autophagy inducer Rapamycin were added to disturb the induction of NETs induced by ox-LDL. The production of NETs marker MPO-DNA and the content of cfDNA/nets in the supernatantwere observed. Meanwhile, Western blot was employed to determine the protein expression of myeloperoxidase (MPO),beclin-l and LC3b in neutrophils. Results Treatment of ox-LDL to adhered neutrophils could lead to the formation of extracellular MPO-DNA and cfDNA/nets generation in a time-and concentration-dependent manner. The protein expression of LC3b,beclin-1 and MPO in neutrophils was up-regulated after treatedwith ox-LDL. TMZ pretreatment significantly inhibited NETs release and reduced the protein expression of LC3b,beclin-1 and MPO in neutrophils,which could be simulated by PI3K/AKT pathway blocker LY294002, while AKT/mTOR inhibitor rapamycin preconditioning did the opposite. Conclusions Ox-LDL-induces the formation of NETs in a time-and concentration-dependent manner. TMZ could down-regulate the level of autophagy and neutrophils, inhibit the induction of ox-LDL on NETs and reduce the release of NETs.