Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis
Objective To investigate the mechanism by which angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) promotes phenotypic switch of vascular smooth muscle cells (VSMCs) and vascular fibrosis through abnormal expression of circadian clock protein BMAL1. Methods Twelve male SD rats (6~8 weeks old,...
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Editorial Office of Journal of Army Medical University
2025-06-01
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| Series: | 陆军军医大学学报 |
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| Online Access: | https://aammt.tmmu.edu.cn/html/202412142.html |
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| author | XUE Lingxia LONG Yaolin FENG Jiayan |
| author_facet | XUE Lingxia LONG Yaolin FENG Jiayan |
| author_sort | XUE Lingxia |
| collection | DOAJ |
| description | Objective To investigate the mechanism by which angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) promotes phenotypic switch of vascular smooth muscle cells (VSMCs) and vascular fibrosis through abnormal expression of circadian clock protein BMAL1. Methods Twelve male SD rats (6~8 weeks old, weighing 180~220 g) were randomly divided into (n=6) a control group and an AT1-AA-positive group [established by active immunization of SD rats with AT1R extracellular loop Ⅱ peptide (AT1R-ECLⅡ)]. HE and Masson stainings were used to observe structural changes and fibrosis in the thoracic aorta (n=3). Western blotting was performed to detect the expression of Collagen I, phenotypic switch-related proteins (SM22, α-SMA, OPN and MMP2) in vascular tissues and primary VSMCs (n=4), as well as the expression of BMAL1 at CT0, CT4, CT8, CT12, CT16, and CT20. Transwell and scratch assays were used to assess the proliferation and migration of VSMCs (n=3). si-RNA was employed to knock down Bmal1, followed by detection of BMAL1, Collagen I, and phenotypic conversion-related protein expression (n=3). Additionally, AT1-AA-positive AT1R-knockout (AT1R-KO) rats were constructed to measure BMAL1 expression in thoracic aortic tissues (n=4). Results The AT1-AA-positive rats had significantly thickened thoracic aortic vessel wall [(140±9)% vs (120±5)%, P<0.05], badly arranged VSMCs, obvious blue Masson staining, and up-regulated Collagen I expression (P<0.05). In the thoracic aorta of AT1-AA-positive rats and AT1-AA-treated VSMCs, the expression of contractile phenotype-related proteins (α-SMA, SM22) was decreased (P<0.05), while the expression of synthetic phenotype-related proteins (OPN, MMP2) was increased (P<0.05). AT1-AA enhanced the scratch healing ability and migration ability of VSMCs. Furthermore, both mRNA and protein levels of Bmal1 were significantly up-regulated at CT12 (P<0.05), and the rhythmicity of Bmal1 was lost. Knockdown of Bmal1 partially ameliorated AT1-AA-induced phenotypic switch of VSMCs. Compared with AT1-AA-positive WT rats, AT1-AA-positive AT1R-KO rats showed significantly reduced BMAL1 expression in the thoracic aorta (1.35±0.06 vs 0.86±0.07，P<0.001). At the cellular level, AT1-AA-induced phenotypic switch and high Collagen I expression in VSMCs were partially improved in AT1R-KO VSMCs. Conclusion AT1-AA promotes VSMCs phenotypic conversion and vascular fibrosis through the AT1R-Bmal1 axis.
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| format | Article |
| id | doaj-art-5fbde8d3df3648d18a22f3181af7d50f |
| institution | OA Journals |
| issn | 2097-0927 |
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| publishDate | 2025-06-01 |
| publisher | Editorial Office of Journal of Army Medical University |
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| series | 陆军军医大学学报 |
| spelling | doaj-art-5fbde8d3df3648d18a22f3181af7d50f2025-08-20T02:22:45ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272025-06-0147111155116410.16016/j.2097-0927.202412142Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosisXUE Lingxia0LONG Yaolin1FENG Jiayan2School of Basic Medical Sciences， Shanxi Medical University， Taiyuan， Shanxi， ChinaSchool of Basic Medical Sciences， Shanxi Medical University， Taiyuan， Shanxi， ChinaSchool of Basic Medical Sciences， Shanxi Medical University， Taiyuan， Shanxi， China Objective To investigate the mechanism by which angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) promotes phenotypic switch of vascular smooth muscle cells (VSMCs) and vascular fibrosis through abnormal expression of circadian clock protein BMAL1. Methods Twelve male SD rats (6~8 weeks old, weighing 180~220 g) were randomly divided into (n=6) a control group and an AT1-AA-positive group [established by active immunization of SD rats with AT1R extracellular loop Ⅱ peptide (AT1R-ECLⅡ)]. HE and Masson stainings were used to observe structural changes and fibrosis in the thoracic aorta (n=3). Western blotting was performed to detect the expression of Collagen I, phenotypic switch-related proteins (SM22, α-SMA, OPN and MMP2) in vascular tissues and primary VSMCs (n=4), as well as the expression of BMAL1 at CT0, CT4, CT8, CT12, CT16, and CT20. Transwell and scratch assays were used to assess the proliferation and migration of VSMCs (n=3). si-RNA was employed to knock down Bmal1, followed by detection of BMAL1, Collagen I, and phenotypic conversion-related protein expression (n=3). Additionally, AT1-AA-positive AT1R-knockout (AT1R-KO) rats were constructed to measure BMAL1 expression in thoracic aortic tissues (n=4). Results The AT1-AA-positive rats had significantly thickened thoracic aortic vessel wall [(140±9)% vs (120±5)%, P<0.05], badly arranged VSMCs, obvious blue Masson staining, and up-regulated Collagen I expression (P<0.05). In the thoracic aorta of AT1-AA-positive rats and AT1-AA-treated VSMCs, the expression of contractile phenotype-related proteins (α-SMA, SM22) was decreased (P<0.05), while the expression of synthetic phenotype-related proteins (OPN, MMP2) was increased (P<0.05). AT1-AA enhanced the scratch healing ability and migration ability of VSMCs. Furthermore, both mRNA and protein levels of Bmal1 were significantly up-regulated at CT12 (P<0.05), and the rhythmicity of Bmal1 was lost. Knockdown of Bmal1 partially ameliorated AT1-AA-induced phenotypic switch of VSMCs. Compared with AT1-AA-positive WT rats, AT1-AA-positive AT1R-KO rats showed significantly reduced BMAL1 expression in the thoracic aorta (1.35±0.06 vs 0.86±0.07，P<0.001). At the cellular level, AT1-AA-induced phenotypic switch and high Collagen I expression in VSMCs were partially improved in AT1R-KO VSMCs. Conclusion AT1-AA promotes VSMCs phenotypic conversion and vascular fibrosis through the AT1R-Bmal1 axis. https://aammt.tmmu.edu.cn/html/202412142.htmlangiotensin ⅱ-1 type 1 receptor autoantibodybmal1angiotensin ⅱ-1 type 1 receptorvascular smooth muscle cellsphenotypic transitionvascular fibrosis |
| spellingShingle | XUE Lingxia LONG Yaolin FENG Jiayan Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis 陆军军医大学学报 angiotensin ⅱ-1 type 1 receptor autoantibody bmal1 angiotensin ⅱ-1 type 1 receptor vascular smooth muscle cells phenotypic transition vascular fibrosis |
| title | Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| title_full | Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| title_fullStr | Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| title_full_unstemmed | Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| title_short | Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| title_sort | angiotensin ii type 1 receptor autoantibody at1r bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis |
| topic | angiotensin ⅱ-1 type 1 receptor autoantibody bmal1 angiotensin ⅱ-1 type 1 receptor vascular smooth muscle cells phenotypic transition vascular fibrosis |
| url | https://aammt.tmmu.edu.cn/html/202412142.html |
| work_keys_str_mv | AT xuelingxia angiotensiniitype1receptorautoantibodyat1rbmal1axispromotesphenotypictransitionofvascularsmoothmusclecellsandvascularfibrosis AT longyaolin angiotensiniitype1receptorautoantibodyat1rbmal1axispromotesphenotypictransitionofvascularsmoothmusclecellsandvascularfibrosis AT fengjiayan angiotensiniitype1receptorautoantibodyat1rbmal1axispromotesphenotypictransitionofvascularsmoothmusclecellsandvascularfibrosis |