Establishment of a transgenic mouse model with coagulation factor V mutation, but with no obvious changes of bone tissue

Wang Yu-ying; Zhang Yan; Qi Jin-wei; Yin Zhao-guang

doi:10.3969/j.issn.2095-4344.2016.46.003

Abstract：BACKGROUND:Blood coagulation factor V gene mutation in non-traumatic femoral head necrosis has been shown to have an higher incidence than that in healthy and secondary non-traumatic femoral head necrosis, and the incidence of thrombosis is positively related. Inactivated blood coagulation factor V can accelerate the activation of prothrombin and the generation of thrombin. Mutations at arg-306, arg-506 and arg-679 wil result in the blood clots and hypercoagulable state. Here, this study is designed to investigate the influence of R506Q/R679Q on osteonecrosis. OBJECTIVE:To establish the mouse model of mutations of Gln506Arg and Gln679Arg in coagulation Factor V (Factor VR506Q/R679Q). METHODS:Factor VR506Q/R679Q point mutation target vector was constructed by molecular cloning technology, the linearization vector was transfected into embryonic stem cel s, and then G418-resistant cel s were screened and used for microinjection. The target blastocysts were transplanted to the fal opian tube of estrus mice to obtain the Chimera mice carrying bilateral LoxP gene, fol owed by mated with CMV-cre transgenic mice, and then the mice with Factor VR506Q/R679Q point mutations were obtained. After genotype identification by PCR, hematoxylin-eosin staining results and percentage of empty lacunae were compared between the mutant and wild-type mice, and rat bone tissue and bone mass were analyzed. RESULTS AND CONCLUSION:There were no obvious abnormalities in the embryonic and postnatal development, percentage of empty lacunae and bone mass of Factor VR506Q/R679Q point mutation mice when compared with the wild-type mice. These results suggest that the mouse model with Factor VR506Q/R679Q point mutation is established successful y, but there is no significant change in the bone tissue. The fol owing research should focus on the effect of external stimulus on the incidence of osteonecrosis in a mutant mouse.