[1]韩永魁,岳云飞,张奎,等.GLP-1(7-36)抑制ApoE-/-小鼠主动脉斑块面积及泡沫细胞内CD36、ACAT1表达及其效应研究[J].第三军医大学学报,2019,41(20):1947-1953.
 HAN Yongkui,YUE Yunfei,ZHANG Kui,et al.GLP-1(7-36) inhibits atherosclerosis progression in ApoE-/- mouse aorta by lowering expressions of CD36 and ACAT1 in foam cells[J].J Third Mil Med Univ,2019,41(20):1947-1953.
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GLP-1(7-36)抑制ApoE-/-小鼠主动脉斑块面积及泡沫细胞内CD36、ACAT1表达及其效应研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第20期
页码:
1947-1953
栏目:
基础医学
出版日期:
2019-10-30

文章信息/Info

Title:
GLP-1(7-36) inhibits atherosclerosis progression in ApoE-/- mouse aorta by lowering expressions of CD36 and ACAT1 in foam cells
作者:
韩永魁岳云飞张奎申晓彧
山西医科大学第二医院心血管内科
Author(s):
HAN Yongkui YUE Yunfei ZHANG Kui SHEN Xiaoyu

Department of Cardiology, Second Hospital of Shanxi Medical University, Taiyuan, Shanxi Province, 030000, China

关键词:
胰升血糖素样肽-1动脉粥样硬化ACAT1CD36
Keywords:
glucagon-like peptide-1 atherosclerosis ACAT1 CD36
分类号:
R329.2;R329.4;R543.1
文献标志码:
A
摘要:

目的 探讨GLP-1(7-36)对高脂饮食喂养的ApoE-/-小鼠动脉粥样硬化的发生发展及巨噬细胞来源的泡沫细胞内CD36、ACAT1表达的影响。方法 ①18~20 g的6~8周龄雄性ApoE-/-基因敲除小鼠实验分4组(n=10):普通饮食组、高脂饮食组、GLP-1(7-36)组、GLP-1(7-36)+Exendin(9-39)组。通过HE染色、油红O染色、免疫组化CD36、ACAT1抗体染色明确小鼠主动脉根部粥样硬化斑块的面积。②巨噬细胞实验分8组(n=4):空白对照组、泡沫细胞组、不同浓度GLP-1(7-36)组、GLP-1(7-36)+Exendin(9-39)组。RT-PCR检测各组细胞CD36、ACAT1 mRNA的表达;Western blot检测各组细胞CD36、ACAT1蛋白的表达;高效液相色谱法检测各组细胞总胆固醇(TC)、游离胆固醇(FC)及胆固醇酯(CE)的含量;细胞免疫荧光分析CD36、ACAT1受体蛋白表达。结果①与空白对照组相比,泡沫细胞组CD36、ACAT1mRNA表达水平较高(P<0.05);与泡沫细胞组相比,GLP-1组CD36、ACAT1mRNA表达水平明显降低(P<0.05);与GLP-1组相比,GLP-1+Exendin组CD36、ACAT1mRNA表达水平较高(P<0.05);与空白对照组相比,泡沫细胞组CD36、ACAT1蛋白表达水平较高(P<0.05);与泡沫细胞组相比,GLP-1组CD36、ACAT1蛋白表达水平明显降低(P<0.05);与GLP-1组相比,GLP-1+Exendin组CD36、ACAT1蛋白表达水平较高(P<0.05);②HE、油红O染色结果显示高脂饮食组斑块面积占主动脉内膜面积比例(30.3±3.8)%;GLP-1组斑块面积占主动脉内膜面积比例(15.1±4.3)%,与高脂饮食组相比较,GLP-1组主动脉斑块明显减少(P<0.05);GLP-1+Exendin组斑块面积占主动脉内膜面积比例(25.7±3.4)%,较GLP-1组内膜下可见大量的粥样斑块(P<0.05);GLP-1(7-36)降低小鼠主动脉CD36、ACAT1阳性细胞百分比表达水平(P<0.05)。与空白对照组相比,泡沫细胞组TC、FC、CE含量较高 ( P<0.05);与泡沫细胞组相比,GLP-1组TC、FC、CE含量明显降低 (P<0.05);与GLP-1组相比,GLP-1+Exendin组TC、FC、CE含量较高( P<0.05)结论 GLP-1通过降低CD36、ACAT1的表达减少细胞内胆固醇的蓄积,抑制巨噬细胞泡沫化,最终达到抑制动脉粥样硬化斑块发生发展。

Abstract:

Objective To investigate whether glucagon-like peptide-1 (GLP-1) (7-36) inhibits the progression of aortic atherosclerotic plaque in ApoE knockout (ApoE-/-) mice fed with high-fat diet and the effect of GLP-1 (7-36) on the expressions of CD36 and acyl-coenzyme A:cholesterol acyltransferase 1 (ACAT1) in macrophage-derived foam cells. MethodsForty male ApoE-/- mice (6-8 weeks old, weighing 18-20 g) were randomized equally into normal diet group, high-fat diet (model) group, GLP-1 (7-36) group, and GLP-1 (7-36)+Exendin (9-39) group. The area of the atherosclerotic plaque in the aortic root of the mice was determined using HE staining, oil red O staining, and immunohistochemical staining with CD36 and ACAT1 antibodies. In the cell experiment, the macrophages (RAW264.7 cells) were induced with oxidized low-density lipoprotein (ox-LDL) and treated with different concentrations of GLP-1 (7-36), alone or in combination with Exendin (9-39). The expressions of CD36 and ACAT1 mRNAs and proteins in the cells was detected using RT-PCR and Western blotting, respectively. The levels of total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) in the cells were detected using high-performance liquid chromatography, and immunofluorescence analysis was performed to detect the expressions of CD36 and ACAT1 receptor proteins. ResultsCompared with the control mice, ApoE-/- mice on high-fat diet showed significantly higher expression levels of CD36 and ACAT1 at both the mRNA and protein levels in the aortic atherosclerotic plaques (P<0.05). CD36 and ACAT1 expressions in the plaques were significantly lowered by treatment of the mice with GLP-1 (7-36) (P<0.05); the expression levels of CD36 and ACAT1 were significantly higher in GLP-1+Exendin group than in GLP-1 group (P<0.05). HE and oil red O staining showed that in the model group, the mean proportion of the plaque area in the aortic intimal area was (30.3±3.8)%, which was significantly lowered to (15.1±4.3)% in GLP-1 group (P<0.05); the ratio of the plaque area was (25.7±3.4)% in GLP-1+Exendin group, where significantly more numerous atherosclerotic plaques were observed below the intima as compared with GLP-1 group (P<0.05). GLP-1 (7-36) significantly decreased the percentage of CD36- and ACAT1-positive cells in the aorta of the mice (P<0.05). ConclusionGLP-1 can reduce the accumulation of intracellular cholesterol by inhibiting the expression of CD36 and ACAT1 to suppress the foaming of macrophages, and thus inhibits the formation of atherosclerotic plaques.
 

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更新日期/Last Update: 2019-10-25