Identification of molecular markers for pre-thrombotic state: validation in the rabbits with tibia fracture or lung cancer

Currently there is no reports on identifying molecular markers for pre-thrombotic state in animal experimental settings. It's necessary to explore the potential molecular markers for pre-thrombotic state to provide evidence for the pharmacotherapy of venous thromboembolism (VTE).

PT, TT, APTT, FIB, D-D and AT-III seem to be sensitive molecular markers for identifying the pre-thrombotic state. CD31, VEGF, ANG and vWF play an important role in the progression of the pre-thrombotic state.

A total of 36 male rabbits were randomly divided into Sham, Tibia fracture, Lung cancer, Tibia fracture and lung cancer group. Pro-thrombin time (PT), thrombin time (TT), activated partial pro-thrombin time (APTT), fibrinogen (FIB), D-dimer (D-D) and anti-thrombin III (AT-III) at baseline, 1, 7, 14 and 21 days after operation were detected. Immuno-histochemical staining of tumor tissue sections was performed to observe the expression of CD31, VEGF and vWF. Western blot and qPCR were used to detect the expression levels of ANG and VEGF gene in tumor tissues. Furthermore, we analyzed the tumor tissue with HE staining, immunochemistry, qPCR and Western blot. all the data were statistically analyzed with GraphPad Prism 6.

There were 0, 1, 2, 4 cases of VTE in sham, tibia fracture, lung cancer, tibia fracture and lung cancer group respectively. The level of PT, APTT, TT and AT-III were significantly decreased after operation (all P<0.05), whereas the level of FIB and D-D were significantly increased after operation (all P<0.05). The level of PT, APTT, TT and AT-III were gradually decreased in tibia fracture, lung cancer, tibia fracture and lung cancer group respectively (all P<0.05), while the level of FIB and D-D were gradually increased in tibia fracture, lung cancer, tibia fracture and lung cancer group respectively (all P<0.05). The medullary lymphocytes increased significantly in the lung cancer group, tibia fracture and lung cancer group (all P<0.05), and the number of medullary lymphocytes in tibia fracture and lung cancer group is significantly higher than that of lung cancer group (P<0.001). The expression of CD31, VEGF and vWF in Tibia fracture and lung cancer group is significantly higher than that of lung cancer group (all P<0.001). The expression ANG and VEGF in tibia fracture and lung cancer group is significantly higher than that of lung cancer group (all P<0.01). Conclusions: PT, TT, APTT, FIB, D-D and AT-III seem to be sensitive molecular markers for identifying the pre-thrombotic state. CD31, VEGF, ANG and vWF play an important role in the progression of the pre-thrombotic state.