›› 2012, Vol. 32 ›› Issue (8): 1056-.doi: 10.3969/j.issn.1674-8115.2012.08.020

• 论著(基础研究) • 上一篇    下一篇

SUMO特异性蛋白酶1在动脉粥样硬化发病机制中的作用

黄 弦1, 吴 际1, 程金科2   

  1. 1.上海交通大学 生命科学技术学院生物化学与分子生物学系, 上海 200240; 2.上海交通大学 基础医学院生物化学与分子细胞生物学系, 上海 200025
  • 出版日期:2012-08-28 发布日期:2012-08-29
  • 通讯作者: 程金科, 电子信箱: jkcheng@shsmu.edu.cn。
  • 作者简介:黄 弦(1987—), 男, 硕士生;电子信箱: g11510910@gmail.com。

Role of SUMO-specific protease 1 in pathogenesis of atherosclerosis

HUANG Xian1, WU Ji1, CHENG Jin-ke2   

  1. 1.Department of Biochemistry and Molecular Biology, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China;2.Department of Biochemistry and Molecular Cellular Biology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2012-08-28 Published:2012-08-29

摘要:

目的 探讨SUMO特异性蛋白酶1(SENP1)在小鼠动脉粥样硬化发病机制中的作用。方法 采用主动脉整体和主动脉根部油红O染色以及主动脉根部巨噬细胞标志物MOMA-2的免疫组织化学染色,观察SENP1+/+ X Apoe-/-(n=5)与SENP1+/- X Apoe-/-(n=4)两组小鼠在饲喂高胆固醇高脂食物后动脉粥样硬化病理改变。采用乙酰化低密度脂蛋白(ac-LDL)诱导巨噬细胞RAW264.7泡沫化,油红O染色检测RAW264.7 si-ns及RAW264.7 si-SENP1的泡沫细胞形成能力。采用Real-Time PCR和Western blotting分别检测RAW264.7 si-ns和RAW264.7 si-SENP1中脂肪酸结合蛋白4(FABP4)mRNA和蛋白的相对表达量。结果 饲喂高胆固醇高脂食物后,油红O染色显示,SENP1+/- X Apoe-/-小鼠的斑块相对面积(斑块面积/血管总面积)和主动脉弓斑块面积均大于SENP1+/+ X Apoe-/-小鼠,分别为(6.716±1.442)%和(5.952±2.332)%以及(364 249±45 838)和(273 486±158 814)μm2;免疫组织化学染色显示,主动脉根部巨噬细胞面积/斑块面积分别为(41.00±0.05)%和(40.47±0.07)%,差异无统计学意义(P=0.954)。Real-Time PCR和Western blotting检测以及油红O染色结果显示,RAW264-7 si-SENP1的FABP4表达水平及泡沫细胞形成能力均较RAW264-7 si-ns高。结论 SENP1通过下调FABP4的表达,抑制巨噬细胞源性泡沫细胞形成,从而抑制动脉粥样硬化的发生。

关键词: 动脉粥样硬化, 泡沫细胞, SUMO特异性蛋白酶1, 脂肪酸结合蛋白4

Abstract:

Objective To investigate the potential role of SUMO-specific protease 1 (SENP1) in the pathogenesis of atherosclerosis in mice. Methods The development of atherosclerosis in SENP1+/+ X Apoe-/- mice (n=5) and SENP1+/- X Apoe-/- mice (n=4) fed with high cholesterol and high fat diet was observed with whole aorta and aorta root oil red O staining and aorta root immunohistochemical staining of macrophage marker MOMA-2. Acetylated low density lipoprotein (ac-LDL) was used to induce the formation of foam cells and oil red O staining, and the capacities of foam cell formation were compared between RAW264.7 si-ns and RAW264.7 si-SENP1. The relative expression of fatty acid-binding protein 4 (FABP4) mRNA and protein in RAW264.7 si-ns and RAW264.7 si-SENP1 was detected by Real-Time PCR and Western blotting. Results Oil red O staining indicated that after feeding with high cholesterol and high fat diet, the relative atherosclerotic lesion areas (lesion areas/total vessel areas) and aorta arch lesion areas of SENP1+/- X Apoe-/- mice were significantly larger than those of SENP1+/+ X Apoe-/- mice [(6.716±1.442) % vs (5.952±2.332)% and (364 249±45 838) μm2 vs (273 486±158 814) μm2]. Immunohistochemical staining revealed that the macrophage area/plaque area in aorta root in SENP1+/- X Apoe-/- mice and SENP1+/+ X Apoe-/- mice were (41.00±0.05)% and (40.47±0.07)% respectively, and there was no significant difference between them (P=0.954). Real-Time PCR, Western blotting and oil red O staining demonstrated that FABP4 level and capacity of foam cell formation of RAW264.7 si-SENP1 were significantly higher than those of RAW264.7 si-ns. Conclusion SENP1 may suppress the expression of FABP4 and capacity of foam cell formation of macrophages, thus may inhibit the development of atherosclerosis.

Key words: atherosclerosis, foam cell, SUMO-specific protease 1, fatty acid-binding protein 4