Zhang Jinhui, Zhang Xili, Liu Xunqiang, Chen Huanjun, Wang Jifeng, Ji Min
Yan'an Hospital Affiliated To Kunming Medical University, Kunming, 650032, China.
First Affiliated Hospital of Kunming Medical University, Kunming, 650032, China.
Cardiovasc Toxicol. 2024 Mar;24(3):302-320. doi: 10.1007/s12012-024-09838-5. Epub 2024 Mar 7.
Abdominal aortic aneurysm (AAA) is a chronic vascular degenerative disease. Vascular smooth muscle cells (VSMCs) are essential for maintaining the integrity of healthy blood vessels. Macrophages play an important role in the inflammatory process of AAA. However, the effect of macrophage-derived exosome LncRNA PVT1 on VSMCs is unclear. Exosomes from M1 macrophages (M1φ-exos) were isolated and identified. The expression of LncRNA PVT1 in M1φ-exos was determined. AAA cell model was constructed by treating VSMCs with Ang-II. AAA cell model was treated with M1φ exosomes transfected with si-LncRNA PVT1 (M1φ-exo). VSMCs were transfected with miR-186-5p mimic and oe-HMGB1. Cell viability was detected by CCK-8. The accumulation of LDH was detected by ELISA. Western blot was used to detect the expression of HMGB1, inflammatory factors (IL-6, TNF-α and IL-1β) and pyroptosis-related proteins (GSDMD, N-GSDMD, ASC, NLRP3, Caspase-1 and Cleaved-Capase-1). Cell pyroptosis rate was detected by flow cytometry. At the same time, the targeting relationship between miR-186-5p and LncRNA PVT1 and HMGB1 was verified by double fluorescein experiment. Exosomes from M1φ were successfully extracted. The expression of LncRNA PVT1 in M1φ-exos was significantly increased. M1φ-exo promotes inflammation and pyroptosis of VSMCs. M1φ-exos inhibited the inflammation and pyroptosis of VSMCs. LncRNA PVT1 can sponge miR-186-5p mimic to regulate HMGB1 expression. MiR-186-5p mimic further inhibited inflammation and pyroptosis induced by M1φ-exos. However, oe-HMGB1 could inhibit the reversal effect of miR-186-5p mimic. LncRNA PVT1 in exosomes secreted by M1φ can regulate HMGB1 by acting as ceRNA on sponge miR-186-5p, thereby promoting cell inflammatory and pyroptosis and accelerating AAA progression.
腹主动脉瘤（AAA）是一种慢性血管退行性疾病。血管平滑肌细胞（VSMC）对于维持健康血管的完整性至关重要。巨噬细胞在AAA的炎症过程中起重要作用。然而，巨噬细胞衍生的外泌体LncRNA PVT1对VSMC的影响尚不清楚。分离并鉴定了来自M1巨噬细胞的外泌体（M1φ-exos）。测定了M1φ-exos中LncRNA PVT1的表达。通过用血管紧张素II处理VSMC构建AAA细胞模型。用转染了si-LncRNA PVT1的M1φ外泌体（M1φ-exo）处理AAA细胞模型。用miR-186-5p模拟物和过表达HMGB1（oe-HMGB1）转染VSMC。通过CCK-8检测细胞活力。通过ELISA检测乳酸脱氢酶（LDH）的积累。蛋白质免疫印迹法用于检测HMGB1、炎症因子（IL-6、TNF-α和IL-1β）和焦亡相关蛋白（GSDMD、N-GSDMD、ASC、NLRP3、Caspase-1和裂解的Caspase-1）的表达。通过流式细胞术检测细胞焦亡率。同时，通过双荧光素酶实验验证miR-186-5p与LncRNA PVT1和HMGB1之间的靶向关系。成功提取了来自M1φ的外泌体。M1φ-exos中LncRNA PVT1的表达显著增加。M1φ-exo促进VSMC的炎症和焦亡。M1φ-exos抑制VSMC的炎症和焦亡。LncRNA PVT1可以吸附miR-186-5p模拟物以调节HMGB1的表达。miR-186-5p模拟物进一步抑制M1φ-exos诱导的炎症和焦亡。然而，oe-HMGB1可以抑制miR-186-5p模拟物的逆转作用。M1φ分泌的外泌体中的LncRNA PVT1可以通过作为竞争性内源RNA（ceRNA）吸附miR-186-5p来调节HMGB1，从而促进细胞炎症和焦亡并加速AAA进展。
