Serpine1 Regulates Peripheral Neutrophil Recruitment and Acts as Potential Target in Ischemic Stroke

Abstract

Zhijun Pu,1– 5 Xinyu Bao,1– 5 Shengnan Xia,1– 5 Pengfei Shao,1– 5 Yun Xu1– 5 1Department of Neurology, Nanjing Drum Tower Hospital, Medical School and the State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, Jiangsu, 210008, People’s Republic of China; 2Institute of Brain Sciences, Nanjing University, Nanjing, Jiangsu, 210093, People’s Republic of China; 3Jiangsu Key Laboratory for Molecular Medicine, Medical School of Nanjing University, Nanjing, Jiangsu, 210008, People’s Republic of China; 4Jiangsu Province Stroke Center for Diagnosis and Therapy, Nanjing, Jiangsu, 210008, People’s Republic of China; 5Nanjing Neurology Clinic Medical Center, Nanjing, Jiangsu, 210008, People’s Republic of ChinaCorrespondence: Yun Xu, Email [email protected]: Peripheral neutrophil infiltration can exacerbate ischemia–reperfusion injury. We focused on the relationship between various peripheral immune cells and cerebral ischemia–reperfusion (I/R) injury.Methods: In this study, we investigated the effects of dauricine on neuronal injury induced by ischemia–reperfusion and peripheral immune cells after ischemic stroke in mouse model, and we explored the undefined mechanisms of regulating peripheral immune cells through RNA sequencing and various biochemical verification in vitro and in vivo.Results: We found that dauricine improved the neurological deficits of I/R injury, reduced the infarct volume, and improved the neurological scores. Furthermore, dauricine reduced the infiltration of neutrophils into the brain after MCAO-R and increased peripheral neutrophils but unchanged the permeability of the endotheliocyte Transwell system in an in vitro blood-brain barrier (BBB) model. RNA sequencing showed that chemotaxis factors, such as CXCL3, CXCL11, CCL20, CCL22, IL12a, IL23a, and serpine1, might play a crucial role. Overexpression of serpine1 reversed LPS-induced migration of neutrophils. Dauricine can directly bind with serpine1 in ligand–receptor docking performed with the Autodock and analyzed with PyMOL.Conclusion: We identified chemotaxis factor serpine1 played a crucial role in peripheral neutrophil infiltration, which may contribute to reduce the neuronal injury induced by ischemia–reperfusion. These findings reveal that serpine1 may act as a potential treatment target in the acute stage of ischemic stroke.Keywords: ischemia–reperfusion injury, dauricine, neutrophil recruitment, serpine1

Keywords

• ischemia–reperfusion injury
• dauricine
• neutrophil recruitment
• serpine1