血三七促骨折愈合活性成分及作用机制研究

Fracture is a common clinical traumatic disease,however,there is no better effective drugs to heal it at present.Polygonum amplexicaule D. Don var.sinense Forbes et Hemsl., a traditional folk medicine in the West of Hubei Province ("Xuesanqi" in Chinese) has been long used to treat fractures and traumatic injury, and it is considered being the best drug to heal fracture.In addition, its distribution is very wide. However,it is not reported about the mechanism of action of Xuesanqi promoting fracture healing,and it is not clear which active ingredients are responsible for so up to now. To solve these problems,the previous investigation on the ethyl acetate fraction of "Xuesanqi" revealed that the fraction promoted the proliferation and differentiation of osteoblast MC3T3-E1. Bioassay-guided fractionation led to the purification of 8 pure compounds from the active fraction. Further bioassay tests on 8 chemical compounds showed the presence of the prelimilary structure-activity relationships (SARs) between chemical structures and promoting fracture healing activity. .This project is mainly designed to find more active ingredients based on promoting osteoblast MC3T3-E1 proliferation screening method, investigate on their SARs, and discover noval leading compounds. Furthermore, detection of the expression level of protein and cytokine about ALP, BGP, BMP, TGF-β1, VEGF and FGF will be performed to elucidate the mechanism of promoting fracture healing by making use of the techniques of cell and molecular biology based on the osteoblast MC3T3-E1 and fracture animal models. Initiation of this project will help to discover new resources of drugs for promoting fracture healing and provide scientific evidence for development of new drugs.

骨折是临床上常见的创伤性疾病，目前尚无疗效较好的药物。血三七是鄂西土家族民间治疗骨折的"要药"，治疗骨折及跌打损伤效果显著,且资源十分丰富。但其活性成分不明确，也尚未见对其促骨折愈合作用机制研究的报道。针对这些问题，申请人前期研究发现其乙酸乙酯部位具有较好地促成骨细胞MC3T3-E1增殖和分化作用，且对该活性部位进行了初步的分离纯化，得到8个单体化合物，它们的结构和活性显示了初步的构效关系。.本项目拟基于促成骨细胞MC3T3-E1增殖筛选模型，进一步发现更多的活性成分，并进行构效关系研究，发现先导化合物。然后以MC3T3-E1细胞和骨折动物为模型，通过使用细胞与分子生物学方法，检测ALP 、BGP、BMP、TGF-β1、VEGF、FGF等蛋白及细胞因子表达水平，阐明其促骨折愈合作用机制。本项目的开展将为发现促骨折愈合新的药物资源奠定基础，为开发新药提供科学依据。

骨折是临床上常见的创伤性疾病，目前尚无疗效较好的药物。血三七是鄂西土家族民间治疗骨折的“要药”，治疗骨折及跌打损伤效果显著,且资源十分丰富。但其活性成分不明确，也尚未见对其促骨折愈合作用机制研究的相关报道。针对这些问题，申请人前期研究发现其乙酸乙酯部位具有较好地促成骨细胞MC3T3-E1增殖和分化作用，在该项的支柱下，对该活性部位和正丁醇部位进行了系统的分离纯化，得到70多个单体化合物，鉴定了其中的59个，有12个新化合物。. 对分离的化合物进行构效关系研究，发现该植物具有的抗肿瘤活性，且抗肿瘤活性物质为总黄酮，其中两种黄酮苷具有抗乳腺癌的活性。以MC3T3-E1细胞和骨折动物为模型，通过使用细胞与分子生物学方法，检测ALP 、PGE2、VEGF、BMP-2、TGF-β等相关因子的表达和蛋白表达情况，证实了血三七具有促骨折愈合的作用且初步阐明了其机理。