CTSS Modulates Stress-Related Carotid Artery Thrombosis in a Mouse FeCl3 Model

Shengnan Xu; Limei Piao; Ying Wan; Zhe Huang; Xiangkun Meng; Aiko Inoue; Hailong Wang; Xueling Yue; Xianglan Jin; Guo-Ping Shi; Masafumi Kuzuya; Xian Wu Cheng

doi:10.1161/ATVBAHA.122.318455

CTSS Modulates Stress-Related Carotid Artery Thrombosis in a Mouse FeCl₃ Model

Arterioscler Thromb Vasc Biol. 2023 Jul;43(7):e238-e253. doi: 10.1161/ATVBAHA.122.318455. Epub 2023 Apr 27.

Authors

Shengnan Xu^{1

2}, Limei Piao^{1

2}, Ying Wan^{1

2}, Zhe Huang³, Xiangkun Meng⁴, Aiko Inoue^{5

6}, Hailong Wang^{1

2}, Xueling Yue^{1

2}, Xianglan Jin^{1

2}, Guo-Ping Shi⁷, Masafumi Kuzuya^{5

6

8}, Xian Wu Cheng^{1

2}

Affiliations

¹ Department of Cardiology and Hypertension (S.X., L.P., Y.W., H.W., X.Y., X.J., X.W.C.), Yanbian University Hospital, Yanji, China.
² Jilin Provincial Key Laboratory of Stress and Cardiovascular Disease (S.X., L.P., Y.W., H.W., X.Y., X.J., X.W.C.), Yanbian University Hospital, Yanji, China.
³ Department of Neurology, University of Occupational and Environmental Health, Kitakyushu, Fukuoka, Japan (Z.H.).
⁴ Department of Vascular Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China (X.M.).
⁵ Community Healthcare & Geriatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan (A.I., M.K.).
⁶ Institute of Innovation for Future Society, Nagoya University, Aichi-ken, Japan (A.I., M.K.).
⁷ Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA (G.-P.S.).
⁸ Meitetsu Hospital, Nagoya, Japan (M.K.).

PMID: 37128920
DOI: 10.1161/ATVBAHA.122.318455

Abstract

Background: Exposure to chronic psychological stress is a risk factor for metabolic cardiovascular disease. Given the important role of lysosomal CTSS (cathepsin S) in human pathobiology, we examined the role of CTSS in stress-related thrombosis, focusing on inflammation, oxidative stress, and apoptosis.

Methods: Six-week-old wild-type mice (CTSS^+/+) and CTSS-deficient mice (CTSS^-/-) randomly assigned to nonstress and 2-week immobilization stress groups underwent iron chloride3 (FeCl₃)-induced carotid thrombosis surgery for morphological and biochemical studies.

Results: On day 14 poststress/surgery, stress had increased the lengths and weights of thrombi in the CTSS^+/+ mice, plus harmful changes in the levels of PAI-1 (plasminogen activation inhibitor-1), ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 13 motifs), and vWF (von Willebrand factor) and arterial tissue CTSS expression. Compared to the nonstressed CTSS^+/+ mice, the stressed CTSS^-/- mice had decreased levels of PAI-1, vWF, TNF (tumor necrosis factor)-α, interleukin-1β, toll-like receptor-4, cleaved-caspase 3, cytochrome c, p16^INK4A, gp91^phox, p22^phox, ICAM-1 (intercellular adhesion molecule-1), MCP-1 (monocyte chemoattractant protein-1), MyD88 (myeloid differentiation primary response 88), and MMP (matrix metalloproteinase)-2/-9 and increased levels of ADAMTS13, SOD (superoxide dismutase)-1/-2, eNOS (endothelial NO synthase), p-Akt (phospho-protein kinase B), Bcl-2 (B-cell lymphoma-2), p-GSK3α/β (phospho-glycogen synthase kinases alpha and beta), and p-Erk1/2 (phospho-extracellular signal-regulated kinase 1 and 2) mRNAs and/or proteins. CTSS deletion also reduced the arterial thrombus area and endothelial loss. A pharmacological inhibition of CTSS exerted a vasculoprotective action. In vitro, CTSS silencing and overexpression, respectively, reduced and increased the stressed serum and oxidative stress-induced apoptosis of human umbilical vein endothelial cells, and they altered apoptosis-related proteins.

Conclusions: CTSS inhibition appeared to improve the stress-related thrombosis in mice that underwent FeCl₃-induction surgery, possibly by reducing vascular inflammation, oxidative stress, and apoptosis. CTSS could thus become a candidate therapeutic target for chronic psychological stress-related thrombotic events in metabolic cardiovascular disease.

Keywords: apoptosis; cathepsin; endothelial cell; oxidative stress; thrombosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cardiovascular Diseases*
Carotid Artery Thrombosis*
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Inflammation / pathology
Mice
Plasminogen Activator Inhibitor 1 / genetics
Thrombosis* / etiology
Thrombosis* / metabolism
von Willebrand Factor / metabolism

Substances

ferric chloride
cathepsin S
von Willebrand Factor
Plasminogen Activator Inhibitor 1