| Objective: Based on the theory of“Stasis removal,regeneration and Bone Union”,this paper discusses the changes and trends of VEGF expression in callus tissues and serum of experimental animals after N-butylphthalide treatment,and whether there is a dose-effect relationship between n-butylphthalide and fracture healing in experimental animals,which provides some experimental and theoretical basis for the clinical application of n-butylphthalide in fracture treatment.Methods: One hundred and fifty male rats were randomly divided into model control group,low dose group(10 mg/kg),middle dose group(25 mg/kg),high dose group(50 mg/kg)and very high dose group(100 mg/kg),the model of middle femoral fracture was made in all rats.The rats in each group were given intragastrically when their vital signs were stable.The model control group was treated with vegetable oil,and the experimental group(low dose group,middle dose group,high dose group,very high dose group)was treated with n-butylphthalide at the same dose for 28 days,0.3 ml/100 g twice daily;Blood samples were taken from 10 rats in each group on the 7th,14 th and 28 th days after operation to examine the contents of calcium,phosphorus,calcium-phosphorus product,Alkaline phosphatase and Vascular endothelial growth factor(VEGF)in serum,and the Vascular endothelial growth factor of VEGF were measured,the animals were killed after x-ray examination.He staining and immunohistochemistry were used to evaluate the growth of callus and angiogenesis in local samples of fracture..Results:1.The results of X-ray examination: X-ray showed that the healing process of fracture and the remodeling speed of callus in the experimental group were better than those in the control group,and the volume of callus was larger.2.Results: on the 7th and 14 th day after operation,the concentrations of serum calcium in the middle-dose group,high-dose group and very high-dose group were lower than those in the model control group and low-dose group(P<0.05),and on the 28 th day after operation,the serum calcium concentrations in the middle-dose group,high-dose group and very high-dose group were lower than those in the model control group and low-dose group(P<0.05),there was no significant difference between the model control group and the experimental group,and the low dose group had higher concentration(P<0.05).3.Results: 7 days after operation,the concentrations of serum phosphorus ion in middle-dose group,high-dose group and very high-dose group were higher than those in model control group and low-dose group(P<0.05),the concentrations in the middle-dose group,high-dose group and very high-dose group were higher than those in the model control group(P<0.05),and there was no significant difference among the three groups at 28 days after operation(P>0.05).4.Results of calcium and phosphorus product: 7 days after operation: compared with the model control group,there were significant differences in calcium and phosphorus product between the experimental group(P>0.05),14 days after operation: there were no significant differences among the groups(P>0.05);28 days after operation: compared with the low-dose group,the middle-dose group,the high-dose group and the very high-dose group had significant difference(P>0.05).5.Results: there was no significant difference in serum Alkaline phosphatase level among the groups 7 days after operation(P>0.05),14 days and 28 days after operation,the concentration of middle-dose group,high-dose group and very high-dose group was higher than that of model control group(P<0.05).6.The results of serum VEGF concentration in each group: 7,14,28 days after operation,compared with the model control group,the experimental group had significant difference(P<0.05),the concentrations in the middle dose group,high dose group and very high dose group were higher than those in the low dose group(P<0.05).7.Results of callus HE staining: granulation tissue was found in the femoral fracture of the model control group and the experimental group,and there were more microvessels in the experimental group than in the control group.On the 14 th day after operation,a lot of new microvessels were found in the callus of the experimental group,while the number of microvessels in the control group was higher than that in the same group on the 7th day after operation.On the 28 th day after operation,the number of microvessels in callus of experimental group and control group decreased.8.Results of VEGF expression in callus: the content of VEGF in callus of experimental group was higher than that of model control group(P<0.05),and reached the peak at 14 days after operation.9.Results of expression of CD31 in callus: the content of CD31 in callus of experimental group was higher than that of control group,and reached the peak at 14 days after operation.Conclusion: N-Butylphthalide can increase the expression of VEGF levels in serum and callus,accelerate the generation of microvascularity,improve local blood supply,and thus promote fracture healing. |
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