| To establish bone-formation of animal model is caused by the cataclasis traction of type2diabetes rats, and to discuss the PPARy, changes after being suffered from fracture during this healing procedure of AP2in type2diabetic rats and the relationship with diabetes cataclasis between healing and obstruction.Methods:To prepare twenty super clean and healthy SD rats are in eight weeks age, making them in two different groups with same numbers by randomly,one group is named experiment, the other is contrast, each group with ten rats.The rats will be feed with high amounts of sugars and fats which is produced specially in experiment group,while the rats will be raised with common feed in the contrast group.After eight weeks,each rat will be evoked by abdominal cavity Injection with STZ(35mg/kg) in experiment group, while other rats will be injected by citrate with the same quantity and same way in contrast group.Two more weeks later, to select SD rats whose blood glucose is higher than16.7mmol/L in empty stomach state, then continue to carry on the following experiment and establish different bone-formation of animal models which are caused by traction of fracturing left tibia in each groups at same time. Then begin to lengthen the left tibia from the first day after the operation,0.3mm/day and twice per day, with the same way to do it till the fourteenth day.But should be stopped lengthening from fifteenth days and to put the rats to death with injection of anesthetics enough which can make them die also in empty state, then to examine the left tibia with X-ray radiography immediately and collect the samples of blood and left tibia under sterile condition.To check cholesterol total, triglycerides, insulin and blood sugar of serum in the eighth, tenth and twelfth weeks after operation, which were collected when put the rat to death. As for the sample of left tibia, we need to take out for HE section-staining, analysis of the number of callus formation of new bone area and bone marrow cavity unit area of fat cells.Western blot and Real-time fluorescence Quantitative detection System for nucleic acid amplification (Real-time PCR Detecting System policy, QPCR) examination.Results:At8weeks, we found that the experimental group rats’ serum triglyceride, total cholesterol, insulin increased significantly compared with control group (P<0.01). After10weeks and12weeks, we found that the experimental group compared with control group in serum triglycerides, total cholesterol, and fasting glucose increased significantly (P<0.01), while serum insulin, no significant difference (P>0.05). Bone tissue section of HE staining showed that the experimental group compared with control group the micro column bone (MCF) to shorten and disordered arrangement and most shallow dye in the initial matrix forefront; Callus formation of new bone area was significantly reduced, while the number of fat cells per unit area in the medullary cavity was increased (P<0.01). X-ray radiography examination showed that the experimental group compared with the control group the formation of callus between the fractures decreased significantly. Western blot and QPCR test results show that the expression of the PPAR gamma and AP2in experimental group compared with control group in callus tissues was increased (P<0.01).Conclusion:Type2diabetes rats fracture traction callus formation disorder. The expression of PPAR gamma and AP2in bone tissue is increase, which may be one of the mechanism lead to fracture healing barrier of type2diabetes. |
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