Effect of transient expression of human cholesteryl ester hydrolase on the cholesterol metabolism in RAW 264.7 macrophage

FENG Xu; et al

doi:10.13406/j.cnki.cyxb.2010.11.007

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Effect of transient expression of human cholesteryl ester hydrolase on the cholesterol metabolism in RAW 264.7 macrophage

更新时间：2024-01-17

- Effect of transient expression of human cholesteryl ester hydrolase on the cholesterol metabolism in RAW 264.7 macrophage
- Journal of Chongqing Medical University Vol. 35, Issue 11, Pages: 1609-1613(2010)
- 作者机构：
  
  辽宁医学院附属第一医院神经外科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.11.007
  CLC： R341
- Published：2010，
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[1]冯旭,罗俊生,关宁,霍晓川,王瑞.人胆固醇酯水解酶在RAW 264.7巨噬细胞内瞬时表达对胆固醇代谢的影响[J].重庆医科大学学报,2010,35(11):1609-1613.

FENG Xu, et al. Effect of transient expression of human cholesteryl ester hydrolase on the cholesterol metabolism in RAW 264.7 macrophage. [J]. Journal of Chongqing Medical University 35(11):1609-1613(2010)
[1]冯旭,罗俊生,关宁,霍晓川,王瑞.人胆固醇酯水解酶在RAW 264.7巨噬细胞内瞬时表达对胆固醇代谢的影响[J].重庆医科大学学报,2010,35(11):1609-1613. DOI： 10.13406/j.cnki.cyxb.2010.11.007.

FENG Xu, et al. Effect of transient expression of human cholesteryl ester hydrolase on the cholesterol metabolism in RAW 264.7 macrophage. [J]. Journal of Chongqing Medical University 35(11):1609-1613(2010) DOI： 10.13406/j.cnki.cyxb.2010.11.007.

摘要

目的:以RAW264.7小鼠单核巨噬细胞为研究对象

探讨人胆固醇酯水解酶（Human cholesteryl ester hydrolase

hCEH）在鼠巨噬细胞内瞬时表达

对细胞内胆固醇代谢产生的影响。方法:利用FuGENE HD转染试剂将pEGFP-N1-hCEH质粒

转染RAW264.7小鼠单核巨噬细胞。24 h后将巨噬细胞与氧化型低密度脂蛋白（Oxidized low density lipoprotein

oX-LDL）（100 mg/L）

共同孵育48 h。荧光显微镜观察及Western blot检测细胞内CEH的表达

油红O染色观察细胞内脂滴堆积情况

高效液相色谱检测细胞内游离胆固醇（Free cholesterol

FC）和胆固醇酯（Cholesterol esters

CE）含量。结果:转染72 h后

荧光细胞数最多

转染率接近40%。Western blot检测显示

hCEH蛋白在鼠巨噬细胞内大量表达并部分抑制鼠胆固醇酯水解酶（Cholesteryl ester hydrolase

CEH）表达。转染hCEH组与ox-LDL诱导组相比

油红O染色阳性细胞数显著减少

细胞内总胆固醇、游离胆固醇和胆固醇酯含量明显下降。结论:hCEH基因在RAW264.7小鼠单核巨噬细胞内成功表达

减少细胞内胆固醇水平

抑制泡沫细胞的形成。

Abstract

Objective:To study the effect of transient expression of human cholesteryl ester hydrolase(hCEH) on the cholesterol metabolism in RAW 264.7 macrophages.Methods:RAW 264.7 macrophages were transfected transiently with pEGFP-N1-CEH under the stimulation of FuGENE HD.After 24 h

macrophages were loaded with oxidized low density lipoprotein(100 mg/L) for 48 h.The expression of pEGFP-N1-CEH was detected with GFP fluorescence and Western blot.The cellular lipid accumulation was examined by oil red O staining.High performance liquid chromatography was used for qualitative and quantitative analysis of intracellular cholesterol and cholesterol esters.Results:There were the most number of fluorescent cells after 72 h.The transfection rate was about 40%.The protein expression level of human CEH significantly increased and partly inhibited expression of mouse CEH protein in mouse macrophage.Compared with ox-LDL group

oil red O-positive cells of hCEH transfection group were greatly reduced.The cellular contents of total cholesterol

free cholesterol and cholesterol ester of hCEH transfection group were also obviously decreased.Conclusion:The gene of hCEH expresses in mouse macrophages cell line RAW 264.7

and inhibits atherosclerotic foam cell formation through reducing intracellular cholesterol level.

关键词

胆固醇酯水解酶氧化低密度脂蛋白泡沫细胞胆固醇代谢

Keywords

Cholesteryl ester hydrolaseOxidized low density lipoproteinFoam cellcholesterol metabolism

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