Role of Rab5 in the formation of macrophage-derived foam cell
Abstract Background Foam cells play a key role in the occurrence and pathogenesis of atherosclerosis. Its formation starts with the ingestion of oxidized low-density lipoprotein (oxLDL). The process is associated with Ras related protein in brain 5 (Rab5) which plays a critical role in regulating en...
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doaj-9a224f043890468387d21617604ac2522020-11-24T21:47:53ZengBMCLipids in Health and Disease1476-511X2017-09-0116111010.1186/s12944-017-0559-6Role of Rab5 in the formation of macrophage-derived foam cellLokwern Chan0Jin Hong1Junjie Pan2Jian Li3Zhichao Wen4Haiming Shi5Jianping Ding6Xinping Luo7Department of Cardiology, Huashan Hospital, Fudan UniversityDepartment of Cardiology, Huashan Hospital, Fudan UniversityDepartment of Cardiology, Huashan Hospital, Fudan UniversityDepartment of Cardiology, Huashan Hospital, Fudan UniversityDepartment of Cardiology, Huashan Hospital, Fudan UniversityDepartment of Cardiology, Huashan Hospital, Fudan UniversityState Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesDepartment of Cardiology, Huashan Hospital, Fudan UniversityAbstract Background Foam cells play a key role in the occurrence and pathogenesis of atherosclerosis. Its formation starts with the ingestion of oxidized low-density lipoprotein (oxLDL). The process is associated with Ras related protein in brain 5 (Rab5) which plays a critical role in regulating endocytosis and early endosomal trafficking. Base on this, we presumed that Rab5 might participate in the maturation of foam cell. The aim of this study is to investigate the effect of Rab5 on macrophage cholesterol during the evolvement of macrophage when induced by oxLDL to the formation of foam cell. Methods Immunohistochemistry was performed to analyze the distribution of macrophages and Rab5 in atherosclerotic plaque. RNA inteference study and transfection of inactive mutant (GFP-Rab5-S34N) and active mutant (GFP-Rab5-Q79L) in U937-derived macrophage were utilized to investigate the impact of Rab5 on the process of macrophage cholesterol, which could be detected by oil red O staining, determination of intracellular lipid content, filipin staining, nile red staining and the costaining of early endosome antigen-1 (EEA-1) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylin dicarbocyanine (Dil)-labelled oxLDL (Dil-oxLDL). Results Rab5 was found abundantly localized in macrophage rich areas of human atherosclerotic lesions. On the foam cell study, the expression of Rab5 was increased after the incubation of oxLDL. The inteference study indicated the depletion of Rab5 led to the decreases of oil red O staining areas, total cholesterol and cholesterol esters in U937-derived marophages. Moreover, the fluorescence intensity of filipin and nile red staining were lower in GFP-Rab5-S34N as compared with GFP-Rab5-Q79L. The confocal study demonstrated less Dil-oxLDL was internalized in GFP-Rab5-S34N as compared with GFP-Rab5-Q79L; the result showed also the decrease in colocalization of internalized Dil-oxLDL and EEA-1 for GFP-Rab5-S34N as compared with GFP-Rab5-Q79L. Conclusions Rab5 plays an important role in modulating the intracellular cholesterol of macrophages and consequently mediating the formation of foam cells.http://link.springer.com/article/10.1186/s12944-017-0559-6RabFoam cellCholesterol |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lokwern Chan Jin Hong Junjie Pan Jian Li Zhichao Wen Haiming Shi Jianping Ding Xinping Luo |
spellingShingle |
Lokwern Chan Jin Hong Junjie Pan Jian Li Zhichao Wen Haiming Shi Jianping Ding Xinping Luo Role of Rab5 in the formation of macrophage-derived foam cell Lipids in Health and Disease Rab Foam cell Cholesterol |
author_facet |
Lokwern Chan Jin Hong Junjie Pan Jian Li Zhichao Wen Haiming Shi Jianping Ding Xinping Luo |
author_sort |
Lokwern Chan |
title |
Role of Rab5 in the formation of macrophage-derived foam cell |
title_short |
Role of Rab5 in the formation of macrophage-derived foam cell |
title_full |
Role of Rab5 in the formation of macrophage-derived foam cell |
title_fullStr |
Role of Rab5 in the formation of macrophage-derived foam cell |
title_full_unstemmed |
Role of Rab5 in the formation of macrophage-derived foam cell |
title_sort |
role of rab5 in the formation of macrophage-derived foam cell |
publisher |
BMC |
series |
Lipids in Health and Disease |
issn |
1476-511X |
publishDate |
2017-09-01 |
description |
Abstract Background Foam cells play a key role in the occurrence and pathogenesis of atherosclerosis. Its formation starts with the ingestion of oxidized low-density lipoprotein (oxLDL). The process is associated with Ras related protein in brain 5 (Rab5) which plays a critical role in regulating endocytosis and early endosomal trafficking. Base on this, we presumed that Rab5 might participate in the maturation of foam cell. The aim of this study is to investigate the effect of Rab5 on macrophage cholesterol during the evolvement of macrophage when induced by oxLDL to the formation of foam cell. Methods Immunohistochemistry was performed to analyze the distribution of macrophages and Rab5 in atherosclerotic plaque. RNA inteference study and transfection of inactive mutant (GFP-Rab5-S34N) and active mutant (GFP-Rab5-Q79L) in U937-derived macrophage were utilized to investigate the impact of Rab5 on the process of macrophage cholesterol, which could be detected by oil red O staining, determination of intracellular lipid content, filipin staining, nile red staining and the costaining of early endosome antigen-1 (EEA-1) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylin dicarbocyanine (Dil)-labelled oxLDL (Dil-oxLDL). Results Rab5 was found abundantly localized in macrophage rich areas of human atherosclerotic lesions. On the foam cell study, the expression of Rab5 was increased after the incubation of oxLDL. The inteference study indicated the depletion of Rab5 led to the decreases of oil red O staining areas, total cholesterol and cholesterol esters in U937-derived marophages. Moreover, the fluorescence intensity of filipin and nile red staining were lower in GFP-Rab5-S34N as compared with GFP-Rab5-Q79L. The confocal study demonstrated less Dil-oxLDL was internalized in GFP-Rab5-S34N as compared with GFP-Rab5-Q79L; the result showed also the decrease in colocalization of internalized Dil-oxLDL and EEA-1 for GFP-Rab5-S34N as compared with GFP-Rab5-Q79L. Conclusions Rab5 plays an important role in modulating the intracellular cholesterol of macrophages and consequently mediating the formation of foam cells. |
topic |
Rab Foam cell Cholesterol |
url |
http://link.springer.com/article/10.1186/s12944-017-0559-6 |
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