中医骨折3期治疗对家兔骨折各种骨痂组织VEGF及VEGF mRNA表达的影响

Effects on VEGF and VEGF mRNA expression in issues of callus in treating fracture of rabbit with three-period treatment theory of bone fracture in traditional Chinese medicine

目的:研究中医骨折3期治则对骨痂VEGF及VEGF-mRNA表达的影响,探讨中药治疗骨折的分期治疗的合理性。方法:动物实验采用双因素一元析因设计方案。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段处,造成3 mm完全骨质缺损,作为骨折愈合模型。采用计算机产生随机数字的方法,将家兔随机平均分为4组:中药3期治疗组、中药1期治疗组、阳性药物对照组以及模型对照组,分别给予3期治疗、1期治疗、阳性药物治疗以及等量蒸馏水。分别于术后3,6,9,14,28,42,56 d每组随机处死5只家兔,取左侧桡骨标本作为观察对象,通过免疫组化和原位杂交方法检查骨折端血肿、纤维骨痂和软骨骨痂中VEGF及VEGF mRNA表达。结果:各中药治疗组对于术后骨折端血肿、纤维骨痂和软骨骨痂VEGF和VEGF mRNA表达均有促进作用,大部分时间点3期治疗组在3种骨痂组织中VEGF及VEGF mRNA表达均有高于其他3组的趋势。结论:中药可以促进骨折端各种骨痂组织VEGF及VEGF mRNA的表达。中药对骨折愈合不同阶段各种组织有不同的作用,针对骨折愈合阶段施以不同方剂的3期组较单一方剂的1期组和阳性药组对骨痂VEGF及VEGF mRNA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 traditional Chinese medicine （TCM） on VEGF and VEGF mRNA expression in the issues of callus of rabbits. And to explore the rationality of phasing method in TCM in treating fracture. Method： one hundred and forty male and healthy rabbits were made 3 mm wide bone defection at lower one third part of both radius as fracture healing model. Then those rabbits were divided into four groups randomly, which are three-period treatment group （TYG）, one-period treatment group （OTG）, positive medicine treatment group （PTG） and model control group （MCG）. Those rabbits in TTG were treated with three-period treatment. Those in OTG were treated with one-period treatment. Those in PTG were feed by guzhecuoshangsan, a Chinese patent medicine which is used to treat bone fracture. Those 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 were selected from every group randomly and were killed by aeroembolism respectively. Their radius were taken out and the left one was taken as the research object. Immunohistochemistry stain and in situ hybridization stain were performed to examinate the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus. Result： All TCM treatment groups can enhance the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus at different time points in fracture healing. The VEGF and VEGF mRNA expression in the three issues of TTG had the tendency of higher than that of the other groups at the most time points after operation. Conclusion： TCM can promote the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus. Different Chinese medicines play various roles on VEGF and VEGF mRNA expression at different stage of fracture healing. Treating fracture in three-period treatment has more predominant effect on the expression of VEGF and VEGF mRNA in the haematoma, fibrous caUus and soft callus than that of treating fracture with single prescription or drug. It is necessary to treat fracture in stages.