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• Original Article •
Experimental MRI study of targeting Rho-associated protein kinase 1 to detect plaques in atherosclerosis
YANG Yawen  XIA Min  SONG Mengxing  MA Zhanlong 

Cite this article as: YANG Y W, XIA M, SONG M X, et al. Experimental MRI study of targeting Rho-associated protein kinase 1 to detect plaques in atherosclerosis[J]. Chin J Magn Reson Imaging, 2024, 15(4): 106-112. DOI:10.12015/issn.1674-8034.2024.04.017.


[Abstract] Objective To observe the expression of Rho-kinase (ROCK) 1 in atherosclerotic plaques, to synthesize a ROCK1-targeted probe and characterize it, then explore its ability to visualize atherosclerotic plaques.Materials and Methods The ROCK1 antibody was coupled with ultra-small superparamagnetic iron oxide nanoparticles to prepare the targeting probe (Fe3O4@PEG-ROCK1), which was characterized and analyzed. Apolipoprotein E knockout (ApoE-/-) mice were fed with high-fat diet and five mice were selected randomly at 10, 16, 22, 28, and 34 weeks, respectively, to measure weight. The expression and activity of ROCK1 were observed by ROCK1 immunostaining and western blot. ApoE-/- mice fed for 34 weeks were divided into two groups, one group (n=10) was injected with Fe3O4@PEG, the other group (n=10) was injected with Fe3O4@PEG-ROCK1. Magnetic resonance imaging was performed before and 8 h and 16 h after injection of the nanoprobe, and the signal intensity of plaques was calculated by Image J software. Abdominal aortic specimens were analyzed by pathology.Results Fe3O4@PEG and Fe3O4@PEG-ROCK1 were uniformly dispersed in aqueous solution, and the hydrated particle sizes were (27.06±1.52) nm and (30.52±2.95) nm, respectively. The Zeta potential was (-35.18±0.31) mV and (-16.60±3.26) mV, respectively. Fe3O4@PEG-ROCK1 could reduce the phagocytosis and clearance of macrophages, was non-toxic within a certain concentration range, and maintains immune activity. The saturation magnetization (0.0868 T) and T2 relaxation rate (162.3 mM-1s-1) indicated that Fe3O4@PEG-ROCK1 had good magnetic sensitivity. With advancing atherosclerosis, the expression of ROCK1 increased (r=0.959, P<0.001). The activity of ROCK1 in abdominal aorta of ApoE-/- mice was higher than that of C57BL/6 mice (0.30±0.02 vs. 0.24±0.02, P<0.01). The results of magnetic resonance imaging showed that compared with plain scan (the plaque signal of Fe3O4@PEG group and Fe3O4@PEG-ROCK1 group were 8.25±1.39 and 7.81±3.22, respectively). After the injection of the probe, the plaque signal of the two groups decreased. Compared with the Fe3O4@PEG group, the plaque signal of the target probe Fe3O4@PEG-ROCK1 group decreased more significantly (8 h, 5.37±1.79 vs. 3.91±2.26, P=0.001; 16 h, 6.68±2.39 vs. 4.61±2.80, P=0.001). Prussian blue staining revealed the deposition of nanoprobes within the plaques, corresponding to the positive areas of immunohistochemistry.Conclusions ROCK1 is highly expressed and active in atherosclerotic plaques. Fe3O4@PEG-ROCK1 may be used as an effective magnetic resonance contrast-enhancing agent for the non-invasive detection of atherosclerotic plaques to promote the visual detection of plaques.
[Keywords] mouse；atherosclerosis；Rho-kinase；nanoprobe；molecular imaging；magnetic resonance imaging

YANG Yawen   XIA Min   SONG Mengxing   MA Zhanlong*  

Department of Radiology, the First Affiliated Hospital of Nanjing Medical University (People's Hospital of Jiangsu Province), Nanjing 210029, China

Corresponding author: MA Z L, E-mail: mazhanlong@126.com

Conflicts of interest   None.

Received  2023-06-01
Accepted  2024-04-07
DOI: 10.12015/issn.1674-8034.2024.04.017
Cite this article as: YANG Y W, XIA M, SONG M X, et al. Experimental MRI study of targeting Rho-associated protein kinase 1 to detect plaques in atherosclerosis[J]. Chin J Magn Reson Imaging, 2024, 15(4): 106-112. DOI:10.12015/issn.1674-8034.2024.04.017.

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