巨噬细胞ADAM8诱导血管平滑肌细胞去分化

Macrophage ADAM8 induced dedifferentiation of vascular smooth muscle cells

目的研究巨噬细胞去整合素金属蛋白酶8(ADAM8)过表达对血管平滑肌细胞去分化的影响。方法利用生物信息学筛选小鼠腹主动脉瘤和人胸主动脉夹层差异表达基因。巨噬细胞转染ADAM8过表达质粒,并收集巨噬细胞培养基处理血管平滑肌细胞。qPCR和Western blotting检测细胞ADAM8表达水平;细胞划痕实验、Edu染色实验分别检测血管平滑肌细胞迁移能力和增殖能力。qPCR检测过表达ADAM8的巨噬细胞培养基对血管平滑肌细胞分化标志基因表达水平的影响。结果ADAM8基因在人胸主动脉夹层和小鼠腹主动脉瘤病变过程中显著上调(P<0.05),且在脂多糖诱导巨噬细胞炎症反应过程中也显著增加(P<0.05)。巨噬细胞过表达ADAM8诱导细胞炎症反应和基质金属蛋白酶(MMP)表达,同时过表达ADAM8的巨噬细胞培养基促进血管平滑肌细胞的迁移和增殖能力,但显著抑制血管平滑肌细胞分化标志基因α-actin和α-tropomyosin的表达水平(P<0.05)。结论巨噬细胞ADAM8促进炎症因子和MMP表达并诱导血管平滑肌细胞的去分化。

Aim To study the effect of macrophage recombinant A disintegrin and metalloprotease 8(ADAM8)overexpression on dedifferentiation of vascular smooth muscle cells.Methods The differentially expressed genes of human aortic dissection and mouse abdominal aortic aneurysm were screened by bioinformatics analysis.ADAM8 was overexpressed in macrophages,and macrophage medium was collected to treat vascular smooth muscle cells.The expression of ADAM8 was detected by qPCR and Western blotting.The migration ability of vascular smooth muscle cells was detected by cell scratch assay.The proliferative ability of vascular smooth muscle cells was evaluated by Edu staining.The effect of macrophage culture medium over expressing ADAM8 on the expression of differentiation marker genes in vascular smooth muscle cells was detected by qPCR.Results The ADAM8 gene was significantly upregulated during the process of human thoracic aortic dissection and mouse abdominal aortic aneurysm lesions(P<0.05),and the expression of ADAM8 was also significantly increased during the inflammatory response of macrophages induced by lipopolysaccharide(LPS)(P<0.05).Overexpression of ADAM8 by macrophages induces cellular inflammatory response and matrix metalloproteinases(MMP)expression.In contrast,overexpressing ADAM8 in macrophage culture medium promotes the migration and proliferation ability of vascular smooth muscle cells,but significantly inhibits the expression of vascular smooth muscle cell differentiation marker genesα-action andα-tropomyosin(P<0.05).Conclusion Macrophage ADAM8 induces dedifferentiation of vascular smooth muscle cells by increasing the expression of inflammatory cytokines and MMP.