Rabbit's Fracture Healing Of Upper Limb In Two-Dimensional Electrophoresis Analysis Of Proteomics

OBJECTIVE Using two-dimensional electrophoresis technology to obtain Two-dimensional electrophoresis map of proteins which callus protein in different stages (2w,4w,8w,12w)of fracture healing and normal bone protein . Observating dynamicly the differences of callus tissue and normal bone in protein species and protein contents, finding differentially expressed proteins. For the subsequent mass spectrometry analysis of proteins,identified specific proteins in fracture healing.METHODS Select 40 New Zealand white rabbits about 2-2.2kg, were divided into 4 groups randomly, which 10 in each group. Establish the fracture model of the rabbit's radial bone, and kill all the animals in 2w, 4w, 8w, 12w later animals respectively . the interception of callus tissue and contralateral. Collected the radius callus tissue and the contralateral normal bone tissue as a control group. Lysis buffer used to extract total protein of bone tissues in each experimental group and the control group. Detected the concentration of protein in each group. Proteins were separated by SDS-PAGE gel electrophoresis and two-dimensional gel electrophoresis。Then it was stained with Coomassie brilliant blue to obtain high-resolution protein mapping。Scanned images by computer software, finally found that the protein differences in the experimental group and control group。RESULTS Two-dimensional gel electrophoresis profiles of the experimental groups and control groups hava high resolution and reproducibility。Protein spots are different in different periods of fracture healing:2w(843±29),4w(727±36),8w(785±30),12w(737±42)。Protein spots in the control group:(710±25)。To the control group as the reference object, The matching protein points of the experimental group have the content of the protein more than 2 or less than 0.5 which it is different proteins。The quantity of the differents protein in all experimental groups is:22(2W),26(4W),24(8W),21(12W)。CONCLUSIONS This experiment established a high-resolution and repeatability of two-dimensional electrophoresis map in the rabbit's fracture healing. Found a number of different proteins associated with the fracture healing. IT is important to further analyzed and identified specific proteins related to fracture healing,and it is useful to clarify the mechanisms of fracture healing of the foundation...