ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages

Background: Carbonic anhydrase I (CA1) has been reported to be a diagnostic and therapeutic target for atherosclerosis (AS). This study aimed to verify the essential role of CA1 in AS progression in CA1-overexpressing mice. Methods: A ApoE [−/−] CA1-overexpressing knock-in mouse model was constructe...

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Main Authors: Jinbao Zong, Changyuan Wang, Hongji Zhou, Yu Song, Kehua Fang, Xiaotian Chang
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Atherosclerosis Plus
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667089525000070
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author Jinbao Zong
Changyuan Wang
Hongji Zhou
Yu Song
Kehua Fang
Xiaotian Chang
author_facet Jinbao Zong
Changyuan Wang
Hongji Zhou
Yu Song
Kehua Fang
Xiaotian Chang
author_sort Jinbao Zong
collection DOAJ
description Background: Carbonic anhydrase I (CA1) has been reported to be a diagnostic and therapeutic target for atherosclerosis (AS). This study aimed to verify the essential role of CA1 in AS progression in CA1-overexpressing mice. Methods: A ApoE [−/−] CA1-overexpressing knock-in mouse model was constructed via CRISPR/Cas9-mediated genome engineering. AS was then induced in these transgenic mice via the administration of a high-fat diet, and a second group simultaneously received treatment with methazolamide (MTZ), a carbonic anhydrase inhibitor. Results: Compared with ApoE [−/−] mice without CA1 overexpression, CA1-overexpressing mice had a greater average body weight, regardless of whether their treatment with MTZ or their AS induction status. Sudan IV, hematoxylin and eosin and Oil Red O staining revealed more plaques and fat deposits in the cardiac aortas of CA1-overexpressing mice than in those of ordinary ApoE−/− mice when AS was induced. Moreover, the atherogenic index; low-density lipoprotein, total cholesterol and triglyceride levels were significantly elevated, and high-density lipoprotein levels were declined in the peripheral blood of CA1-overexpressing mice than in that of ordinary ApoE [−/−] mice, regardless of whether these animals were induced to AS. Immunohistochemistry, Von Kossa staining and fluorescence immunohistochemistry revealed increases in CA1 expression, calcium deposition and M1 macrophages in the aortic tissues of CA1-overexpressing mice with AS. MTZ treatment significantly suppressed AS pathologies in the above experiments. Conclusion: These findings revealed aggravated AS in ApoE [−/−] CA1-overexpressing mice and suggest that CA1 aggravates AS by increasing M1-type macrophages, a proinflammatory macrophage subtype.
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spelling doaj-art-6cf41d11277b46eeb1a968e5899cbe852025-08-20T02:54:39ZengElsevierAtherosclerosis Plus2667-08952025-06-016061910.1016/j.athplu.2025.03.003ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophagesJinbao Zong0Changyuan Wang1Hongji Zhou2Yu Song3Kehua Fang4Xiaotian Chang5Clinical Laboratory and Qingdao Key Laboratory of Immunodiagnosis, Qingdao Hiser Hospital Affiliated of Qingdao University (Qingdao Traditional Chinese Medicine Hospital), Renmin Road 4, Qingdao, 266000, PR ChinaDepartment of Dermatology, Qingdao Hospital, University of Health and Rehabilitation Sciences(Qingdao Municipal Hospital), Dengyun Road 369, Qingdao, 266000, PR ChinaMedical Research Center, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, 266000, PR China; Department of Cardiology, Fushun Municipal Central Hospital, Xincheng Road 5, Fushun, Liaoning, 113006, PR ChinaMedical Research Center, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, 266000, PR ChinaClinical Laboratory, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, Shandong, 266000, PR China; Corresponding author. Clinical Laboratory, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, Shandong, 266000, PR ChinaMedical Research Center, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, 266000, PR China; Corresponding author. Medical Research Center, The Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, 266000, PR China.Background: Carbonic anhydrase I (CA1) has been reported to be a diagnostic and therapeutic target for atherosclerosis (AS). This study aimed to verify the essential role of CA1 in AS progression in CA1-overexpressing mice. Methods: A ApoE [−/−] CA1-overexpressing knock-in mouse model was constructed via CRISPR/Cas9-mediated genome engineering. AS was then induced in these transgenic mice via the administration of a high-fat diet, and a second group simultaneously received treatment with methazolamide (MTZ), a carbonic anhydrase inhibitor. Results: Compared with ApoE [−/−] mice without CA1 overexpression, CA1-overexpressing mice had a greater average body weight, regardless of whether their treatment with MTZ or their AS induction status. Sudan IV, hematoxylin and eosin and Oil Red O staining revealed more plaques and fat deposits in the cardiac aortas of CA1-overexpressing mice than in those of ordinary ApoE−/− mice when AS was induced. Moreover, the atherogenic index; low-density lipoprotein, total cholesterol and triglyceride levels were significantly elevated, and high-density lipoprotein levels were declined in the peripheral blood of CA1-overexpressing mice than in that of ordinary ApoE [−/−] mice, regardless of whether these animals were induced to AS. Immunohistochemistry, Von Kossa staining and fluorescence immunohistochemistry revealed increases in CA1 expression, calcium deposition and M1 macrophages in the aortic tissues of CA1-overexpressing mice with AS. MTZ treatment significantly suppressed AS pathologies in the above experiments. Conclusion: These findings revealed aggravated AS in ApoE [−/−] CA1-overexpressing mice and suggest that CA1 aggravates AS by increasing M1-type macrophages, a proinflammatory macrophage subtype.http://www.sciencedirect.com/science/article/pii/S2667089525000070Atherosclerosis (AS)Carbonic anhydrase I (CA1)CalcificationMethazolamide (MTZ)M1-type macrophages
spellingShingle Jinbao Zong
Changyuan Wang
Hongji Zhou
Yu Song
Kehua Fang
Xiaotian Chang
ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
Atherosclerosis Plus
Atherosclerosis (AS)
Carbonic anhydrase I (CA1)
Calcification
Methazolamide (MTZ)
M1-type macrophages
title ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
title_full ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
title_fullStr ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
title_full_unstemmed ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
title_short ApoE [−/−] CA1-overexpressing knock-in mice aggravated atherosclerosis by increasing M1 macrophages
title_sort apoe ca1 overexpressing knock in mice aggravated atherosclerosis by increasing m1 macrophages
topic Atherosclerosis (AS)
Carbonic anhydrase I (CA1)
Calcification
Methazolamide (MTZ)
M1-type macrophages
url http://www.sciencedirect.com/science/article/pii/S2667089525000070
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