Cathepsin K deficiency prevented stress-related thrombosis in a mouse FeCl3 model
Xueying Jin, Xueling Yue, Zhe Huang, Xiangkun Meng, Shengnan Xu, Yuna Wu, Ying Wan, Aiko Inoue, Megumi Narisawa, Lina Hu, Guo‐Ping Shi, Hiroyuki Umegaki, Toyoaki Murohara, Yanna Lei, Masafumi Kuzuya, Xian Wu Cheng
Abstract
Abstract Background Exposure to chronic psychological stress (CPS) is a risk factor for thrombotic cardiocerebrovascular diseases (CCVDs). The expression and activity of the cysteine cathepsin K (CTSK) are upregulated in stressed cardiovascular tissues, and we investigated whether CTSK is involved in chronic stress-related thrombosis, focusing on stress serum-induced endothelial apoptosis. Methods and results Eight-week-old wild-type male mice (CTSK +/+ ) randomly divided to non-stress and 3-week restraint stress groups received a left carotid artery iron chloride3 (FeCl 3 )-induced thrombosis injury for biological and morphological evaluations at specific timepoints. On day 21 post-stress/injury, the stress had enhanced the arterial thrombi weights and lengths, in addition to harmful alterations of plasma ADAMTS13, von Willebrand factor, and plasminogen activation inhibitor-1, plus injured-artery endothelial loss and CTSK protein/mRNA expression. The stressed CTSK +/+ mice had increased levels of injured arterial cleaved Notch1, Hes1, cleaved caspase8, matrix metalloproteinase-9/-2, angiotensin type 1 receptor, galactin3, p16 IN4A , p22phox, gp91 phox , intracellular adhesion molecule-1, TNF-α, MCP-1, and TLR-4 proteins and/or genes. Pharmacological and genetic inhibitions of CTSK ameliorated the stress-induced thrombus formation and the observed molecular and morphological changes. In cultured HUVECs, CTSK overexpression and silencing respectively increased and mitigated stressed-serum- and H 2 O 2 -induced apoptosis associated with apoptosis-related protein changes. Recombinant human CTSK degraded γ-secretase substrate in a dose-dependent manor and activated Notch1 and Hes1 expression upregulation. Conclusions CTSK appeared to contribute to stress-related thrombosis in mice subjected to FeCl 3 stress, possibly via the modulation of vascular inflammation, oxidative production and apoptosis, suggesting that CTSK could be an effective therapeutic target for CPS-related thrombotic events in patients with CCVDs.