Pretreatment with Eupatilin Attenuates Inflammation and Coagulation in Sepsis by Suppressing JAK2/STAT3 Signaling Pathway

Abstract

Yilun Lu,1– 3,* Ding Li,1– 3,* Yueyue Huang,1– 3 Yuanyuan Sun,1– 3 Hongmin Zhou,1– 3 Fanrong Ye,1– 3 Hongjing Yang,1– 3 Tingting Xu,1– 3 Shichao Quan,3– 6 Jingye Pan1– 5 1Department of Intensive Care Unit, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 2Key Laboratory of Intelligent Treatment and Life Support for Critical Diseases of Zhejiang Province, Wenzhou, People’s Republic of China; 3Wenzhou Key Laboratory of Critical Care and Artificial Intelligence, Wenzhou, People’s Republic of China; 4Collaborative Innovation Center for Intelligence Medical Education, Wenzhou, People’s Republic of China; 5Zhejiang Engineering Research Center for Hospital Emergency and Process Digitization, Wenzhou, People’s Republic of China; 6Department of General Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jingye Pan, Department of Intensive Care Unit, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China, Email [email protected] Shichao Quan, Department of General Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China, Email [email protected]: Sepsis is an aggressive and life-threatening organ dysfunction induced by infection. Excessive inflammation and coagulation contribute to the negative outcomes for sepsis, resulting in high morbidity and mortality. In this study, we explored whether Eupatilin could alleviate lung injury, reduce inflammation and coagulation during sepsis.Methods: We constructed an in vitro sepsis model by stimulating RAW264.7 cells with 1 μg/mL lipopolysaccharide (LPS) for 6 hours. The cells were divided into control group, LPS group, LPS+ Eupatilin (Eup) group, and Eup group to detect their cell activity and inflammatory cytokines and coagulation factor levels. Cells in LPS+Eup and Eup group were pretreated with Eupatilin (10μM) for 2 hours. In vivo, mice were divided into sham operation group, cecal ligation and puncture (CLP) group and Eup group. Mice in the CLP and Eup groups were pretreated with Eupatilin (10mg/kg) for 2 hours by gavage. Lung tissue and plasma were collected and inflammatory cytokines, coagulation factors and signaling were measured.Results: In vitro, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and tissue factor (TF) expression in LPS-stimulated RAW264.7 cells was downregulated by Eupatilin (10μM). Furthermore, Eupatilin inhibited phosphorylation of the JAK2/STAT3 signaling pathway and suppressed p-STAT3 nuclear translocation. In vivo, Eupatilin increased the survival rate of the mice. In septic mice, plasma concentrations of TNF-α, IL-1β and IL-6, as well as TF, plasminogen activator inhibitor 1 (PAI-1), D-dimer, thrombin-antithrombin complex (TAT) and fibrinogen were improved by Eupatilin. Moreover, Eupatilin alleviated lung injury by improving the expression of inflammatory cytokines and TF, fibrin deposition and macrophage infiltration in lung tissue.Conclusion: Our results revealed that Eupatilin may modulate inflammation and coagulation indicators as well as improve lung injury in sepsis via the JAK2/STAT3 signaling pathway.Keywords: Eupatilin, sepsis, inflammation, coagulation, JAK2/STAT3

Keywords

• eupatilin
• sepsis
• inflammation
• coagulation
• jak2/stat3