Protease nexin-1 deficiency increases mouse hindlimb neovascularisation following ischemia and accelerates femoral artery perfusion

Sonia Selbonne,Véronique Arocas,Marie-Christine Bouton,Celina Madjene,Benjamin Salmon,Yacine Boulaftali

doi:10.1038/s41598-021-92794-9

Abstract

We previously identified the inhibitory serpin protease nexin-1 (PN-1) as an important player of the angiogenic balance with anti-angiogenic activity in physiological conditions. In the present study, we aimed to determine the role of PN-1 on pathological angiogenesis and particularly in response to ischemia, in the mouse model induced by femoral artery ligation. In wild-type (WT) muscle, we observed an upregulation of PN-1 mRNA and protein after ischemia. Angiography analysis showed that femoral artery perfusion was more rapidly restored in PN-1−/− mice than in WT mice. Moreover, immunohistochemistry showed that capillary density increased following ischemia to a greater extent in PN-1−/− than in WT muscles. Moreover, leukocyte recruitment and IL-6 and MCP-1 levels were also increased in PN-1−/− mice compared to WT after ischemia. This increase was accompanied by a higher overexpression of the growth factor midkine, known to promote leukocyte trafficking and to modulate expression of proinflammatory cytokines. Our results thus suggest that the higher expression of midkine observed in PN-1- deficient mice can increase leukocyte recruitment in response to higher levels of MCP-1, finally driving neoangiogenesis. Thus, PN-1 can limit neovascularisation in pathological conditions, including post-ischemic reperfusion of the lower limbs.

Highlights

We previously identified the inhibitory serpin protease nexin-1 (PN-1) as an important player of the angiogenic balance with anti-angiogenic activity in physiological conditions
We used the mouse model of hindlimb ischemia induced by femoral artery ligation in WT and PN-1−/− mice and analysed muscles after 3, 5, 7 and 14 days
We investigated the role of the anti-angiogenic serpin PN-1 in the regulation of the post-ischemic angiogenic response in the mouse model of hindlimb ischemia

Summary

Introduction

We previously identified the inhibitory serpin protease nexin-1 (PN-1) as an important player of the angiogenic balance with anti-angiogenic activity in physiological conditions. We aimed to determine the role of PN-1 on pathological angiogenesis and in response to ischemia, in the mouse model induced by femoral artery ligation. Most serpins are powerful specific inhibitors of extracellular serine proteases controlling biological processes such as blood coagulation, fibrinolysis, tissue remodelling, and inflammation Their role as central regulators of normal physiological pathways is well illustrated by disease states due to serpin deficiency in humans and by the profound effects induced by genetic mutations in some serpins. As PN-1 strongly limits physiological angiogenesis and is expressed by inflammatory cells[7], we postulate that it could modulate PAD outcome In this context, we used a mouse model of hindlimb ischemia induced by ligation of the femoral artery. This study identifies PN-1 as a possible target to improve muscle perfusion in PAD

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