Forsythiaside B ameliorates coagulopathies in a rat model of sepsis through inhibition of the formation of PAD4-dependent neutrophil extracellular traps

Wenju He; Qiang Xi; Huantian Cui; Pingping Zhang; Rui Huang; Taihuan Wang; Dongqiang Wang

doi:10.3389/fphar.2022.1022985

Forsythiaside B ameliorates coagulopathies in a rat model of sepsis through inhibition of the formation of PAD4-dependent neutrophil extracellular traps

Front Pharmacol. 2022 Nov 2:13:1022985. doi: 10.3389/fphar.2022.1022985. eCollection 2022.

Authors

Wenju He¹, Qiang Xi², Huantian Cui³, Pingping Zhang¹, Rui Huang⁴, Taihuan Wang⁴, Dongqiang Wang¹

Affiliations

¹ Department of Integration of Traditional Chinese and Western Medicine, First Central Hospital Affiliated to Nankai University, Tianjin First Central Hospital, Tianjin, China.
² Department of Practice and Education, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
³ Shandong Provincial Key Laboratory of Animal Cell and Developmental Biology, School of Life Sciences, Shandong University, Qingdao, China.
⁴ Department of Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin, China.

Abstract

Forsythiaside B (FTB) is one of the main components of Forsythia suspensa (Thunb.) Vahl and exerts anti-inflammatory and anti-oxidative effects. However, its mechanism of action as a treatment for sepsis remains unclear. In this study, we developed a rat model of sepsis using cecal ligation and puncture (CLP) to investigate the effects of FTB on sepsis-associated coagulopathies. Using rats with sepsis, we investigated the effects of FTB on neutrophil extracellular trap (NETs) formation and peptidylarginine deiminase 4 (PAD4) expression in neutrophils. NET (DNase1) and PAD4 (Cl-amidine) inhibitors were used to further investigate whether FTB mitigates sepsis-associated coagulopathies by inhibiting PAD4-dependent NETs production. Our results showed that treatment with FTB increased the survival rate, ameliorated the CLP-induced inflammatory response and multiple organ dysfunction, and reduced CLP-induced pathological changes. FTB also alleviated the associated coagulopathies. Additionally, we demonstrated that treatment with FTB inhibited NETs formation and downregulated PAD4 expression in peripheral neutrophils. The effects of FTB on coagulopathies were similar to those of monotherapy with NET or PAD4 inhibitors. In conclusion, our study confirmed that FTB can alleviate coagulopathies in rats with sepsis. The underlying mechanism of FTB's effect consists in inhibition of PAD4-dependent NETs formation.

Keywords: PAD4; coagulation disorders; forsythiaside B; neutrophil extracellular trap; sepsis.