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. 2024 Jan 16;14:1438. doi: 10.1038/s41598-024-51996-7

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Sustained ketosis downregulates CCR2 content and inhibit its downstream signals. (A) Pro MMP9 levels at week 1 in AAA tissue of SD and KDp rats (11.3 ± 2 × 10³ vs. 9.5 ± 2.1 × 10³; p = ns). (B) Active MMP9 levels in AAA tissue of SD and KDp rats (2.1 ± 1.4 × 10³ vs. 0.9 ± 0.7 × 10³; p = 0.04). (C) Total MMP2 levels in AAA tissue of SD and KDp rats (7 ± 0.7 × 10³ vs. 1.8 ± 1 × 10³; p < 0.001). Pro and active MMP9 and total MMP2 levels were measured by integrated optical density (IOD). (D) Representative zymogram from AAA tissue homogenates at week 1 demonstrating pro and active MMP-9 and total MMP-2 levels in SD and KDp rats (E) MMP9/TIMP1 complex levels in AAA tissue of SD and KDp rats (6.3 ± 5 × 10² vs. 15 ± 8.7 × 10²; p = 0.02). ELISA of AAA tissue homogenates in SD and KDp rats provided levels of (G) Total MMP9 (5 ± 1.1 × 10^–1 vs. 6 ± 0.6 × 10^–1; p = ns), (H) SD AAA tissue harvested at day 6 (week 1) stained with VVG at 10× and 5× magnification respectively. TIMP 1 (5.9 ± 1.6 × 10³ vs. 3.4 ± 3 × 10³; p = 0.03). (I) KD AAA tissue harvested at day 6 (week 1) stained with VVG at 10× and 5× magnification respectively. (J) AAA VVG-staining quantification for SD and KDp at week 1 (7 ± 2 vs. 26 ± 18 respectively; p = 0.03). (K) TGFβ-1 protein content expressed as a ratio to Caveolin 1 content in AAA tissue from SD and KDp rats at week 1 (2.2 ± 0.8 × 10^–1 vs. 5.7 ± 3.4 × 10^–1; p = 0.2). (L) TGFβ-1 protein content expressed as a ratio to Caveolin 1 content in AAA tissue from SD and KDp rats at week 2 (0.3 ± 0.1 vs. 0.5 ± 0.1; p = ns). (M and N) Trichrome staining of abdominal aortas (cross-sectional) with 5× magnification and 10× magnification in SD and KDp rats to visualize collagen deposition in animal aortic tissue. (O) AAA collagen staining quantification for SD and KDp at week 2 (20 ± 4 vs. 32 ± 6; p = 0.006). (P) Collagen 1 protein content expressed as a ratio to GAPDH content in AAA tissue at week 2 for SD and KDp rats (1.5 ± 1.0 × 10^–2 vs. 10 ± 8.2 × 10^–2, p = 0.03). (Q) α-SMA protein content expressed as a ratio to GAPDH content in AAA tissue of SD and KDp rats (3.4 ± 1 vs. 3.8 ± 0.7; p = ns). (R) NF Kappa β protein content expressed as a ratio to Caveolin I content in AAA tissue of SD and KDp rats (0.06 ± 0.03 vs. 0.03 ± 0.01; p = ns). (S) Representative Western blots of collagen 1, NF Kappa β, α-SMA, TGFβ-1, GAPDH and Caveolin 1 in AAA tissue. Data presented as mean ± standard deviation. ns > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001 using Student’s t test.