Objective To investigate the effect and mechanism of Yaobitong Capsules (腰痹通胶囊, YBT) on deep vein thrombosis (DVT). Methods Multi-scale evaluation of the effect of YBT was conducted using a rat lower limb deep vein ligation model, a zebrafish blood stasis model induced by phenylhydrazine, and an oxygen glucose deprivation (OGD)-induced human umbilical vein endothelial cell (HUVEC) injury model. In the rat lower limb deep vein ligation model, the ratio of thrombus weight to length and hematoxylin-eosin (HE) staining were used to evaluate the pathological changes of thrombus. Six coagulation parameters, plasma tissue factor (TF), endothelin-1 (ET-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), hypoxia inducible factor-1α (HIF-1α) and other indicators were detected. In the zebrafish blood stasis model, the stasis of red blood cells and platelet aggregation were assessed via o-dianisidine staining and confocal microscopy, respectively. qRT-PCR was employed to measure the expressions of coagulation-related genes (fga, fgb, fgg, f2). In the OGD induced HUVEC cell injury model, qRT-PCR, Western blotting and immunofluorescence were used to detect the expressions of TF, VCAM-1, ET-1, HIF-1α, as well as the phosphorylation and nuclear translocation of nuclear factor-κB (NF-κB) p65. Results In the rat model of deep vein ligation of the lower limbs, YBT could reduce the ratio of thrombus weight to length, improve histopathological changes (P < 0.05), prolong activated partial thromboplastin time (APTT) (P < 0.001), reduce fibrinogen (FIB), D-dimer, fibrinogen and fibrin degradation products (FDP), TF, ET-1, sVCAM-1, IL-6, IL-1β, TNF -α, MDA and HIF-1α levels (P < 0.01, 0.001), and increase SOD activity (P < 0.05, 0.01). In the zebrafish blood stasis model, YBT could significantly increase cardiac erythrocyte staining intensity and circulating platelet number (P < 0.001), and down-regulate the expressions of coagulation related genes (P < 0.01, 0.001). In the OGD induced HUVEC cell injury model, YBT could down-regulate the mRNA expressions of TF, VCAM-1, ET-1 and HIF-1α (P < 0.05, 0.01, 0.001), down-regulate the protein expressions of TF and HIF-1α (P < 0.05, 0.01, 0.001), inhibit the phosphorylation and nuclear translocation of NF-κB p65 (P < 0.05, 0.01). Conclusion YBT may effectively inhibit DVT by inhibiting HIF-1α/NF-κB p65/TF axis (down-regulating HIF-1α expression, inhibiting NF-κB p65 phosphorylation and nuclear translocation, thereby reducing TF overexpression), alleviating the hypoxia inflammation cascade, protecting endothelial function, and improving hypercoagulability.

Yaobitong Capsules  /  deep vein thrombosis  /  coagulation  /  hypoxia  /  inflammation  /  HIF-1α/NF-κB p65/TF axis