骨折三期治疗对骨生长因子FGF-2表达影响的实验研究

Experimental study of effects of the three-phase treatment in TCM for fracture on FGF-2 expression during fracture healing in rats

目的 :观察骨折三期治疗对FGF 2在骨折愈合过程中表达的影响。方法 :16 8只雄性SD大鼠随机取 6只作为正常对照组 ,其余SD大鼠双胫骨上 1/ 3段前部的中点造成 2mm× 1 5mm的缺损。术后随机平均分为模型对照组、一期治疗组和三期治疗组 ,每组于不同的时间点分别处死 6只 ,取材 ,常规组织学检查 ,免疫组化法检测标本中的FGF 2 ,应用计算机图像分析系统进行定量分析。结果 :①缺损区内血肿细胞、炎性细胞、间充质细胞、成骨细胞、新形成骨细胞FGF 2表达呈强阳性 ,成熟骨细胞阴性或微弱阳性 ;②骨折后三组FGF 2表达阳性细胞数和平均光密度值均即明显增高 ,但是 ,中药治疗组显著高于模型对照组 (P <0 0 5 ) ,三期治疗组有高于一期治疗组的趋势 ,FGF 2表达高峰出现时间中药治疗组显著早于模型组 ,三期治疗组较一期治疗组提前了 80 %。结论 :中药治疗能显著促进骨折愈合过程中FGF 2的表达 ,三期治疗明显强于一期治疗 ,因此对骨折进行分期辨证治疗是非常必要的。

Objective: To observe the influence of three-phase treatment in TCM for fracture on FGF-2 expression during fracture healing in rats and to probe the effect of three-phase treatment for fracture in TCM and its mordern medical mechanism. Methods: Among 168 male SD rats,6 rats were taken randomly as normal group.In another rats,2 mm×1 5 mm defect model were made at the middle of the anterior part of the up-shaft of tibia.At first,6 male SD rats Postoperatively,162 male SD rats were divided randomly into three groups:the model control group,the one-phase treatment group and the three-phase treatment group,and then every group was divided randomly into 9 small groups,6 rats in each small group.The rats in one-phase treatment group underwent TCM in 1 phase,those in three-treatment group in 3 phases and those in model control group saline at the same dose.On 1,3,5,7,9,12,15,21,28 days after operation,a small group in each group were killed respectively.Specimens were prepared and examinated histologically in routine.The expression of FGF-2 was detected by immunohistochemical staining and quantitatively analysed by computer image analysis system. Results: ①In different time points after operation,there was positive expression of FGF-2 in cells in hematom,inflammatory cells,mesenchymal cells,osteoblasts and immature osteocytes in defect area while mature osteocytes showed negative or slightly positive.②The mean optical density of the expression of FGF-2 obviously went up postoperatively,gradually increasing with development of fracture repair,reaching quickly peak during the formation of primary callus,and then rapidly decreasing,and returning by degrees to the normal level with remodelling of callus.However,mean optical density in the two group treated in TCM was significantly higher than that in the model control group( P <0 05).Further,mean optical density in the three-phase treatment group should a highter tendence than that in the one-phase treatment group.The occurring time of peak of FGF-2 expression in the three-phase treatment went ahead of that of one-phase treatment at 80%.Change tendence of positive cell number was generally consistent with that of mean optical density. Conclusion: TCM can greatly promote FGF-2 expression in fracture healing,and the three-phase treatment is notablely stronger than the one-phase treatment in accelerating FGF-2 expression.Its very necessary to treat fracture with phases.