Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
Stem Cell Rev Rep. 2022 Mar;18(3):1041-1053. doi: 10.1007/s12015-021-10180-4. Epub 2021 May 12.
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.
钙结合蛋白 1（STC-1）是一种分泌性糖蛋白，参与炎症、细胞凋亡和坏死的调节。我们研究了过表达 STC-1 的脂肪干细胞（ADSC）来源的外泌体对损伤颈动脉内膜的再内皮化作用。
ADSC 用含有前 STC-1 的慢病毒载体转染。PHK-26 作为分子探针，用于追踪被小鼠动脉内皮细胞（MAEC）吞噬的外泌体。通过划痕试验和管形成试验验证 STC-1-ADSC-Exosome（S-ADSC-Exo）在 MAEC 中的作用。通过 Western blot 和定量逆转录聚合酶链反应检测 STC-1 和 NLRP3 炎性小体的表达。用靶向 STC-1 的 siRNA 的拮抗剂抑制再内皮化作用。用导丝插入颈总动脉管腔诱导颈动脉内膜机械损伤。取颈动脉进行组织学检查、免疫荧光染色和 Evan's 蓝染色。
STC-1 转染显著提高了 ADSC、其外泌体和 MAEC 中的 STC-1 水平。与对照组和 ADSC-Exo 组相比，富含 STC-1 的外泌体显著抑制了 MAEC 中 NLRP3、Caspase-1 和 IL-1β的表达，表现出良好的侧向迁移能力，并促进血管生成。用靶向 STC-1 的 siRNA 处理完全消除了 STC-1 对 NLRP3、Caspase-1 和 IL-1β 的下调作用，并抑制了 S-ADSC-Exo 对侧向迁移和血管生成的作用。体内给予 S-ADSC-Exo 对损伤后的颈动脉内膜具有再内皮化作用，表现为动脉壁较薄、NLRP3 炎性小体表达较低和更多存活的内皮细胞。
通过基因修饰外泌体使其包含高水平的 STC-1，ADSC 来源的外泌体对损伤颈动脉内膜的再内皮化作用增强，可能通过抑制 NLRP3 炎性小体介导的炎症。
