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Updated: Oct 1, 2025
Quantification of Atherosclerosis in Mice
Published on: June 12, 2019
The LOX-1 receptor ectopically expressed in the liver alleviates atherosclerosis by clearing Ox-LDL from the
Zhiwen Wang1, Juan Chen1, Zhuanglin Zeng2
1Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Objective:
Oxidized Low-Density-Lipoprotein (Ox-LDL) is the core factor in the development of atherosclerosis. However, there are few therapies aimed at eliminating Ox-LDL. Here in this study, we investigate whether the ectopically expression of the lectin-like oxidized low density lipoprotein receptor (LOX-1) in the liver could lead to the elimination of circulating Ox-LDL and prevent the deposition in the vascular wall, thereby alleviating the progression of atherosclerosis.
Methods:
Apolipoprotein E-deficient (ApoE-/-) mice were randomly divided into three groups, the control group, the AAV8-TBG-eGFP group (eGFP group) and AAV8-TBG-LOX-1 group (LOX-1 group). In the LOX-1 group, mice received an injection of virus dilution AAV8-TBG-LOX-1 (1.16 × 1011 virus genome (v.g)/animal/100 μl). The mice in the control group and eGFP group received the same amount of sterile saline and AAV8-TBG-eGFP virus dilution injections. The expression of LOX-1 in the liver was detected by immunofluorescent, western blot and immunohistochemistry. The safety of the virus was assessed by hematoxylin-eosin (H&E) staining, blood biochemical analyses and immunofluorescent. The function of LOX-1 in the liver was detected by the co-localization of LOX-1 and Dil-labeled Ox-LDL (Dil-Ox-LDL) under laser scanning confocal microscope. The extent of Ox-LDL in plasma was detected by ELISA. Changes in blood lipids were assessed through blood biochemical analysis. The progression of atherosclerotic lesions was detected by Oil red O staining. And the expression of Vascular Cell Adhesion Molecule-1 (VCAM-1) in endothelial cells and the extent and migration of macrophages in atherosclerotic plaque were detected by immunofluorescence staining. The protein expression in liver was assessed by qRT-PCR and western blot.
Results:
The expression of LOX-1 was stable in liver within 4 weeks. Ectopically expressed LOX-1 in the liver phagocytosed and degraded Ox-LDL and reduced Ox-LDL from circulation but did not have a significant effect on blood lipid levels. After the expression of LOX-1 in liver, Ox-LDL can be cleared by the hepatocytes, thereby reducing VCAM-1 expression in vascular endothelium and the migration of macrophages in plaques, and eventually alleviating the progression of atherosclerosis. Functional expression of LOX-1 in hepatocytes may facilitate the metabolic clearance of Ox-LDL by upregulating the expression of ATP-binding cassette G5 and G8 (ABCG5/G8), which is the primary neutral sterol transporter in hepatobiliary and transintestinal cholesterol excretion.
Conclusion:
Ectopic liver-specific expression of LOX-1 receptor alleviates the progression of atherosclerosis by clearing Ox-LDL from circulation.
Insights
Introducing lectin-like oxidized low density lipoprotein receptor (LOX-1) into the liver clears circulating oxidized low-density lipoprotein (Ox-LDL). This approach reduces atherosclerosis progression by preventing Ox-LDL deposition in blood vessels.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Atherosclerosis Research
Background:
- Oxidized low-density lipoprotein (Ox-LDL) is a key driver of atherosclerosis.
- Current therapies targeting Ox-LDL elimination are limited.
- Understanding mechanisms to clear Ox-LDL is crucial for developing new treatments.
Purpose of the Study:
- To investigate the potential of ectopically expressed lectin-like oxidized low density lipoprotein receptor (LOX-1) in the liver for eliminating circulating Ox-LDL.
- To determine if liver-specific LOX-1 expression can prevent Ox-LDL deposition in vascular walls and alleviate atherosclerosis progression.
Main Methods:
- Apolipoprotein E-deficient (ApoE-/-) mice were used and divided into control, eGFP, and LOX-1 groups.
- Adeno-associated virus serotype 8 (AAV8) vectors were employed for liver-specific gene delivery.
- Assays included immunofluorescence, western blot, ELISA, blood lipid analysis, Oil red O staining, and immunohistochemistry to assess LOX-1 expression, Ox-LDL levels, and atherosclerotic lesion development.
Main Results:
- Stable ectopic expression of LOX-1 in the liver was achieved within 4 weeks.
- Liver LOX-1 effectively phagocytosed and degraded Ox-LDL, reducing circulating levels without significantly altering blood lipids.
- Reduced VCAM-1 expression and macrophage migration in atherosclerotic plaques were observed, indicating alleviated atherosclerosis.
Conclusions:
- Ectopic liver-specific expression of LOX-1 receptor alleviates atherosclerosis progression.
- This is achieved by enhancing the metabolic clearance of circulating Ox-LDL via hepatocytes.
- Upregulation of hepatobiliary sterol transporters (ABCG5/G8) may contribute to this clearance mechanism.
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