外敷中药复方对促进骨折愈合的体外实验研究

Creating a Herbal Formula for Topical Treatment of Bone Fractures

摘要:

骨折损伤至愈合是一个包括多种细胞和组织代谢的复杂过程，可被多种因素干扰。各种原因导致的骨折延迟愈合、不愈合约占骨折病人的5－7％。寻找有效的方法加速损伤愈合一直是骨科研究的热点。传统医学运用中药治疗骨折损伤，历史悠久，疗效显著。但是在有关方剂的药效和机理方面一直缺乏系统的研究。本课题旨在利用现代生物模型，验证中药方剂对骨折愈合的影响，为中医接骨理论提供科学依据和筛选新型方剂。根据中医骨伤的组方原则，本研究从抗炎镇痛，活血化瘀和接骨续筋三类中药中各选出两味最常用的中药，采用均匀设计法将六位中药按照不同剂量配比，组成五条方剂。选择炎症因子(NO)释放、人体静脉上皮细胞(HUVEC-12)和成骨细胞(UMR-106)增殖试验分别反映抗炎镇痛，活血化瘀和接骨续筋的功效，测试各方剂对骨折愈合的影响。结果表明，五组复方体外试验均表现出单重或多重促进骨折愈合的作用。其中五号复方（大黄30克，栀子，三七，红花各9克，接骨木30克，续断15克）显示出全面的抑制NO释放，促进HUVEC-12和UMR-106增殖的活性。研究结果证明了中药复方能从多个方面加速骨折愈合，其主效应包括抗炎镇痛，促进骨折局部的血液循环和新骨再生。研究提供了以大黄、栀子为解毒君药，以三七、红花活血化瘀作臣药，续断、接骨木为佐使的基本外敷复方，实际应用时可根据辩证观察作适当增减，达到最佳疗效。

Abstract:

Fracture healing is a process that involves the proliferation and differentiation of cells and tissues and can be stimulated by drugs. Traditional medicine has a reputation in the therapy on bone fracture. It is believed that herbal formulae promotes bone repair through multiple functions of “anti-inflammation”, “promoting blood circulation” and “stimulating bone formation”. This study aims to engineer a herbal formula under the guidance of Chinese medicine theories, then define the effective dosage ratio between the herbs and demonstrate the biological effects of the formulae on different parameters of healing using relevant in vitro assays. Six herbs which are frequently used for fractures were combined into five formulae according to the “Uniform Design” method. Assays of nitric oxide (NO) inhibition in mouse macrophages, human umbilical vein endothelial cells (HUVEC) and rat osteoblasts proliferation were used to reflect the “anti-inflammation inflammatory”, “circulation promotion” and “bone formation stimulation” functions. The results showed that all the formulae expressed significant activities of NO inhibition, and HUVEC cell and osteoblasts proliferation. Formula 5 with the largest doses (containing 30g Radix et Rhizoma Rhei, 9g Fructus Gardeniae, 9g Radix et Rhizoma Notoginseng, 9g Flos Carthami, 30g Ramulus Sambucus Willamsii, 15g Radix Dipsaci) demonstrated the best results in all the three assays.