背景:慢性心理应激(CPS)是血栓性心脑血管疾病(CCVD)的危险因素。半胱氨酸组织蛋白酶K(CTSK)的表达和活性在应激心血管组织中上调,我们调查了CTSK是否与慢性应激相关的血栓形成有关,重点研究应激血清诱导的内皮细胞凋亡。
结果:8周龄野生型雄性小鼠(CTSK+/+)随机分为非应激和3周束缚应激组,在特定时间点接受左颈动脉氯化铁3(FeCl3)诱导的血栓形成损伤,以进行生物学和形态学评估。在压力/受伤后的第21天,压力增加了动脉血栓的重量和长度,除了血浆ADAMTS13、血管性血友病因子的有害改变,和纤溶酶原激活抑制剂-1,加上损伤动脉内皮丢失和CTSK蛋白/mRNA表达。应激CTSK+/+小鼠损伤动脉裂解Notch1、Hes1、caspase8、基质金属蛋白酶-9/-2、血管紧张素1型受体水平升高,半乳糖苷3,p16IN4A,p22phox,gp91phox,细胞内粘附分子-1,TNF-α,MCP-1和TLR-4蛋白和/或基因。CTSK的药理和遗传抑制改善了应激诱导的血栓形成以及观察到的分子和形态变化。在培养的HUVEC中,CTSK过表达和沉默分别增加和减轻了与凋亡相关蛋白变化相关的应激血清和H2O2诱导的凋亡。重组人CTSK以剂量依赖性方式降解γ-分泌酶底物并激活Notch1和Hes1表达上调。
结论:CTSK似乎有助于FeCl3应激小鼠的应激相关血栓形成,可能是通过血管炎症的调节,氧化产生和凋亡,提示CTSK可能是CCVD患者CPS相关血栓事件的有效治疗靶点.
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.
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 (FeCl3)-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, p16IN4A, p22phox, gp91phox, 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 H2O2-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 FeCl3 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.