抗β2GPI抗体促进巨噬细胞摄取oxLDL加速动脉粥样硬化进展

崔佳; 仇鹏; 赵培; 贾敏

doi:10.13898/j.cnki.issn.1000-2200.2023.03.006

抗β2GPI抗体促进巨噬细胞摄取oxLDL加速动脉粥样硬化进展

Anti-β2GPⅠ antibody promotes uptake of oxLDL by macrophages to accelerate the progression of atherosclerosis

摘要

摘要:
目的研究抗β2糖蛋白Ⅰ(β2GPⅠ)抗体在apoE基因敲除(apoE^-/-)小鼠巨噬细胞摄取功能及动脉粥样硬化之间的关系。
方法将雄性apoE^-/-小鼠随机分为抗β2GPI抗体组和模型组，每组8只，均给予高脂饲料喂养8周，其中，抗β2GPI抗体组小鼠腹腔注射抗β2GPI抗体，模型组小鼠腹腔注射同等剂量0.9%氯化钠溶液稀释的同源对照IgG抗体。8周后解剖小鼠，分离小鼠主动脉根组织，行Movat染色、免疫组织化学染色，观察主动脉根部动脉粥样硬化斑块的病理组织学特点；Western blotting检测主动脉斑块中内质网应激和巨噬细胞的标志蛋白表达；腹腔巨噬细胞体外培养，抗β2GPI抗体组和模型组分别加入氧化型低密度脂蛋白(oxLDL)+抗β2GPI抗体、oxLDL刺激24 h，观察巨噬细胞摄取oxLDL情况。
结果Movat染色显示，抗β2GPI抗体组小鼠主动脉斑块增多，斑块面积较大，胶原纤维、平滑肌纤维含量均减少，与模型组相比差异有统计学意义(P < 0.05~P < 0.01)。免疫组织化学染色显示，抗β2GPI抗体组小鼠主动脉根部斑块巨噬细胞面积大于模型组(P < 0.05)。Western blotting结果显示，抗β2GPI抗体组小鼠中内质网应激通路标志蛋白IRE1-α、p-IRE1-α、ATF6、GRP-94及巨噬细胞CD36蛋白表达量均高于模型组(P < 0.05~P < 0.01)。荧光染色结果显示，抗β2GPI抗体组绿色荧光强度明显强于模型组(P < 0.01)。泡沫细胞油红O染色结果显示，抗β2GPI抗体组存在泡沫细胞且数量明显多于模型组(P < 0.01)。
结论抗β2GPI抗体促进apoE^-/-小鼠巨噬细胞摄取oxLDL，加速泡沫细胞的形成，并且通过内质网应激促进CD36在巨噬细胞中的表达，加速摄取oxLDL，进而促进动脉粥样硬化进展。

Abstract:
ObjectiveTo investigate the role of anti-β2-glycoprotein Ⅰ(β2GPⅠ)antibody in the relationship between macrophage uptake and atherosclerosis in apoE knockout (apoE^-/-) mice.
MethodsThe male apoE^-/- mice were randomly divided into the anti-β2GPⅠ antibody group and model group, with 8 mice in each group, and fed with high-fat diet for 8 weeks.The mice in the anti-β2GPⅠ antibody group were intraperitoneally injected with anti-β2GPⅠantibody, and the mice in the model group were intraperitoneally injected with the same dose of homologous control IgG antibody diluted with 0.9% sodium chloride solution.After 8 weeks, the mice were dissected, and the aortic root tissues were isolated for Movat staining and immunohistochemical staining to observe the histopathological characteristics of atherosclerotic plaques in the aortic root; Western blotting was used to detect the expression of marker proteins of endoplasmic reticulum stress and macrophage in aortic plaques; the peritoneal macrophages were cultured in vitro, the anti-β2GPⅠ antibody group and model group were stimulated with oxidized low-density lipoprotein (oxLDL) plus anti-β2GPⅠ antibody and oxLDL for 24 hours, respectively, to observe the uptake of oxLDL by macrophages.
ResultsMovat staining showed that the aortic plaque increased, the plaque area was larger, and the contents of collagen fibers and smooth muscle fibers were decreased in the anti-β2GPⅠ antibody group compared with the model group (P < 0.05 to P < 0.01).Immunohistochemical staining indicated that the area of macrophages in the aortic root plaque in the anti-β2GPⅠ antibody group was larger than that in the model group (P < 0.05).The results of Western blotting displayed that the expression levels of marker proteins of endoplasmic reticulum stress pathway such as IRE1-α, p-IRE1-α, ATF6, GRP-94 and macrophage CD36 protein were higher than those in the model group (P < 0.05 to P < 0.01).The results of fluorescence staining showed that the green fluorescence intensity in the anti-β2GPⅠ antibody group was significantly stronger than that in model group (P < 0.01).The results of oil red O staining of foam cells showed that foam cells existed in the anti-β2GPⅠ antibody group and the number of foam cells was significantly higher than that in the model group (P < 0.01).
ConclusionsThe anti-β2GPⅠ antibody promotes the uptake of oxLDL by macrophages in apoE^-/- mice, accelerates the formation of foam cells, and increases the expression of CD36 in macrophages through endoplasmic reticulum stress to accelerate the uptake of oxLDL, thus promoting the progression of atherosclerosis.

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