目的:研究中医骨折三期治则对家兔桡骨骨折愈合过程中内外骨膜及骨髓组织VEGF及VEGFmRNA表达的影响,探讨中药治疗骨折的分期治疗的合理性。方法:140只雄性新西兰白兔,无菌状态下手术,在家兔双侧桡骨下1/3段,造成3mm完全骨质缺损的骨折愈合模型。采用计算机产生随机数字的方法,将家兔随机均分为4组:中药三期治疗组、中药一期治疗组、阳性药物对照组以及模型对照组,分别给予三期治疗、一期治疗、阳性药物治疗以及等量蒸馏水。于术后3、6、9、14、28、42、56d每组随机处死5只家兔,取左侧桡骨标本作为观察对象,采用免疫组化和原位杂交方法检查外骨膜、内骨膜和骨髓组织中VEGF及VEGFmRNA表达,通过图像分析测量各种组织中VEGF及VEGFmRNA表达强度。结果:各中药治疗组对于术后骨折端外骨膜、内骨膜和骨髓组织VEGF和VEGFmRNA表达均有促进作用,三期治疗组有高于其他三组的趋势(P〈0.05)。结论:中药分期治疗对骨折愈合过程中外骨膜、内骨膜和骨髓组织VEGF及VEGFmRNA表达的促进作用更加明显,骨折分期治疗是必要的。
Objective:To observe the effects of the three period treatment theory of bone fracture in TCM(Traditional Chi- nese Medicine) on VEGF and VEGF mRNA expression in the issues of outer periosteum,endosteum and bone marrow of rab- bits,and to explore the rationality of phasing method in TCM in treating fracture. Methods:3 mm bone defection were made at lower one third part of both radius in 140 male healthy rabbits. The rabbits were randomly divided into four groups,including three period treatment group(TTG),one period treatment group(OTG),positive medicine treatment group (PTG) and model control group(MCG). The rabbits in TTG were treated with three period treatment,rabbits in OTG were treated with one period treatment,rabbits in PTG were feed by Guzhe Cuoshangsan(a Chinese patent medicine which was used to treat bone fracture), rabbits in model control group were given no prescription or drug but distilled water as same dose as that of other groups. At day 3,6,9,14,28,42 and 56,five rabbits from every group were randomly selected and were killed by aeroembolism. The left ra- diuses were taken out as the research object. Immunohistochemistry stain and in situ hybridization stain were performed to ex- aminate the VEGF and VEGF mRNA expression in the outer periosteum,endosteum and bone marrow. Results:The VEGF and VEGF mRNA expression of all TCM treatment groups were enhanced in the outer periosteum,endosteum and bone marrow at different time points in fracture healing. The VEGF and VEGF mRNA expression in the three tissues of TTG had the tendency of higher than that of the other groups at the most time points after operation. Conclusion:Treating fracture in stages has more predominant effect on the expression of VEGF and VEGF mRNA in the outer periosteum,endosteum and bone marrow than that of treating fracture with single prescription or drug.