基础医学与临床 ›› 2021, Vol. 41 ›› Issue (2): 219-224.

• 研究论文 • 上一篇    下一篇

沉默CXCL12抑制ox-LDL诱导小鼠巨噬细胞系RAW264.7泡沫化和凋亡

庄红, 张苏川*, 刘慧地, 徐亚宁, 黄璐, 代天, 蒋品   

  1. 江汉大学附属医院(武汉市第六医院) 心血管内科, 湖北 武汉 430015
  • 收稿日期:2020-02-28 修回日期:2020-05-30 出版日期:2021-02-05 发布日期:2021-01-19
  • 通讯作者: *zhangsuchuan@163.com

Silenced CXCL12 inhibits ox-LDL-induced foaming and apoptosis in mouse macrophage cell line RAW264.7

ZHUANG Hong, ZHANG Su-chuan*, LIU Hui-di, XU Ya-ning, HUANG Lu, DAI Tian, JIANG Pin   

  1. Department of Cardiovascular Medicine, Affiliated Hospital of Jianghan University(Wuhan Sixth Hospital), Wuhan 430015, China
  • Received:2020-02-28 Revised:2020-05-30 Online:2021-02-05 Published:2021-01-19
  • Contact: *zhangsuchuan@163.com

摘要: 目的 探讨CXC趋化因子配体12(CXCL12)对氧化低密度脂蛋白(ox-LDL)诱导小鼠腹腔巨噬细胞系RAW264.7泡沫化和凋亡的作用及其机制。方法 用60 mg/L的ox-LDL诱导RAW264.7泡沫化,记为ox-LDL组,不添加ox-LDL的作为对照(NC)组;将si-con和si-CXCL12转染至RAW264.7细胞中再用60 mg/L的ox-LDL处理,分别记为ox-LDL+si-con组、ox-LDL+si-CXCL12组。油红O染色法检测细胞泡沫化; Western blot检测ABCA-1、SRB-1、cleaved caspase-3、cleaved caspase-9、CXCL12、p38MAPK和p-p38MAPK蛋白表达;流式细胞计量术检测细胞凋亡;RT-qPCR检测CXCL12 mRNA表达。结果 ox-LDL处理的RAW264.7细胞体积增大,形状变为圆形,胞质中可明显见到大量红色脂滴;ABCA-1和SRB-1表达水平显著升高(P<0.05),cleaved caspase-3和cleaved caspase-9表达水平显著升高(P<0.05),细胞凋亡率显著升高(P<0.05),CXCL12表达水平显著升高(P<0.05),p-p38MAPK表达水平显著升高(P<0.05)。沉默CXCL12,大部分细胞内未见红染脂滴,仅少数细胞内有部分脂滴,ABCA-1和SRB-1表达水平显著降低(P<0.05),cleaved caspase-3和cleaved caspase-9表达水平显著降低(P<0.05),细胞凋亡率显著降低(P<0.05),p-p38MAPK表达水平显著降低(P<0.05)。结论 沉默CXCL12抑制ox-LDL诱导巨噬细胞RAW264.7泡沫化和细胞凋亡,其可能与p38MAPK信号通路有关。

关键词: CXC趋化因子配体 12, 氧化低密度脂蛋白, 巨噬细胞泡沫化, 凋亡

Abstract: Objective To investigate the effect and mechanism of CXC chemokine ligand 12 (CXCL12) on oxidized low density lipoprotein (ox-LDL)-induced foaming and apoptosis of macrophages. Methods RAW264.7 was incubated with 60 mg/L of ox-LDL to induce foaming of macrophages, which was recorded as ox-LDL group, and to compare with the control without ox-LDL; si-con and si-CXCL12 were transfected to RAW264.7 cells and thenincubated with ox-LDL at 60 mg/L as ox-LDL+si-con group and ox-LDL+si-CXCL12 group, respectively. Oil Red O staining was used to detect cellular foaming; Western blot was used to detect ABCA-1, SRB-1, cleaved caspase-3, cleaved caspase-9, CXCL12, p38 mitogen-activated protein kinase (p38MAPK), and phosphorylated p38MAPK (p-p38MAPK) protein expression; flow cytometry is used to detect apoptosis; CXCL12 mRNA expression was measured by real-time fluorescence quantitative PCR(RT-qPCR). Results The size of ox-LDL-treated macrophages increased, the shape became round, and a large number of red lipid droplets were found in the cytoplasm; the expression of ABCA-1 and SRB-1 and cleaved caspase-3 as well as cleaved caspase-9 were all significantly increased; the apoptosis rate and the expression of CXCL12 and p-p38MAPK were significantly increased(P<0.05). Silencing CXCL12, no red-stained lipid droplets were found in most cells, only a few cells had partial lipid droplets, the expressions of ABCA-1 and SRB-1 were significantly reduced, and the expression of cleaved caspase-3 and cleaved caspase-9, apoptosis rate and the expression of p-p38MAPK significantly reduced(P<0.05). Conclusions Silencing CXCL12 inhibits ox-LDL-induced foaming and apoptosis of mouse macrophage cell line RAW264.7, which may be related to the p38MAPK signaling pathway.

Key words: CXC chemokine ligand 12, oxidized low density lipoprotein, macrophage foaming, apoptosis

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