Exosomes from Adipose Mesenchymal Stem Cells Overexpressing Stanniocalcin-1 Promote Reendothelialization After Carotid Endarterium Mechanical Injury.

Kun Liu,Xiaoyu Wu,Huihua Shi,Xinwu Lu,Weimin Li,Zhiyou Peng

doi:10.1007/s12015-021-10180-4

Kun Liu, Xiaoyu Wu + Show 4 more

Open Access

https://doi.org/10.1007/s12015-021-10180-4

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Abstract

Stanniocalcin-1 (STC-1) is a secreted glycoprotein that participates in the regulation of inflammation, apoptosis, and necrosis. We investigated the reendothelialization effect of exosomes from adipose stem cells (ADSC) overexpressing STC-1 on injured carotid endarterium. ADSCs were transfected with lentivirus vectors containing pre-STC-1. PHK-26 as molecular probe was used to track the exosomes engulfed by mice arterial endothelial cells (MAEC). The role of STC-1-ADSC-Exosome (S-ADSC-Exo) in MAECs was verified through scratch test and tube forming. Expressions of STC-1 and NLRP3 inflammasome were detected by western blot and quantitative reverse transcription polymerase chain reaction. Reendothelialization effect was inhibited by the antagonist of siRNA targeting STC-1. Carotid endarterium mechanical injury was induced by insertion with a guidewire into the common carotid artery lumen. Carotid arteries were harvested for histological examination, immunofluorescence staining, and Evan's blue staining. Transfection of STC-1 significantly enhanced STC-1 levels in ADSCs, their exosomes, and MAECs. Compared with the control group and the ADSC-Exo group, STC-1 enriched exosomes markedly inhibited the expressions of NLRP3, Caspase-1, and IL-1β in MAECs, exhibited good lateral migration capacity, and promoted angiogenesis. Administration of siRNA targeting STC-1 completely abolished down-regulation of NLRP3, Caspase-1, and IL-1β by STC-1 and inhibited effects of S-ADSC-Exo on lateral migration and angiogenesis. In vivo administration of S-ADSC-Exo had reendothelialization effect on post-injury carotid endarterium as evidenced by thinner arterial wall, low-expressed NLRP3 inflammasome, and more living endothelial cells. The reendothelialization effect of exosomes from ADSCs on post-injury carotid endarterium could be enhanced by genetic modification of the exosomes to contain elevated STC-1, possibly through suppression of NLRP3 inflammasome-mediated inflammation.

Highlights

18–25 % ischemic strokes are caused by extracranial carotid atherosclerotic stenosis[1]
Exosomes derived from adipose stem cells (ADSC) were membrane vesicles (Fig. S2 c) and there is no difference in morphology between ADSCExo and S-ADSC-Exo
ADSC-Exosomes engulfed by mice arterial endothelial cells (MAEC) and expressions of STC-1 after transfection

Summary

Introduction

18–25 % ischemic strokes are caused by extracranial carotid atherosclerotic stenosis[1]. For patients at high risk of surgical complications, combination carotid angioplasty with stent implantation has become an accepted alternative to endarterectomy[2]. Post-injury restenosis just like wound-healing comprises complex pathophysiological mechanisms consisting of inflammation, proliferation, and migration followed by remodeling of arterial wall[4]. Reendothelialization is a crucial repair process that endothelial cells rapidly attempt to cover the denuded area by migrating over distance and increasing in number in response to any injury to endothelium[6]. Delayed reendothelialization and impaired endothelial function after mechanical injury are linked to stent thrombosis. Inhibition of inflammation, restoration of endothelial cell function to accelerate reendothelialization process are effective approaches to prevent and treat restenosis after endovascular treatment

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