激素性股骨头坏死过程中血管形成与细胞凋亡的相关性及破瘀通络补肾法干预作用

the pathogenesis key of avascular necrosis of the femoral head is ischemia, but the specific pathogenesis is not too clear.Research states clearly that the steroid-induced avascular necrosis of the femoral head(SANFH) exist over the osteoblast and bone cell apoptosis,being the result of joint action of cell apoptosis and necrosis ;promoting the formation of blood vessels and improving blood supply femoral head are helpful for the restoration of the necrosis.But the relation of the formation of blood vessels and cell apoptosis as well as the research on the series of protein CD34 etc of control the formation of blood vessels have not been seen.This is reported as the breakthrough point in the project,the steroid and endotoxin are used to establish rabbit SANFH model，the model rabbits are differently intervened with "PO YU TONG LUO"and "BU SHEN TONG LUO" function,the formation of blood vessels,new bone formation,CD34、CYR61、CTGF、VEGF expression and VEGFmRNA expression and apoptosis and bcl-2 expression as well as the relationships and the influence of traditional chinese medicine are detected and observed in the process of avascular necrosis of the femoral head by histology, TUNEL and molecular biology technology etc. The Purpose is to further study the pathogenesis of SANFH from the relationship of the formation of blood vessels and cell apoptosis , and explore the mechanism of traditional chinese medicine for SANFH ,and provide the basis for the treatment of SANFH and steroid-induced bone disorders with traditional Chinese medicine .

股骨头坏死的病机关键是缺血，但具体发病机理尚不太清楚。研究表明激素诱导的股骨头缺血性坏死（SANFH）存在过度成骨细胞和骨细胞凋亡，是细胞凋亡和坏死共同作用的结果；促进血管形成，改善股骨头血液供应，有利于坏死的修复。但血管形成与细胞凋亡是否相关以及对调控血管形成的系列蛋白CD34等的研究尚未见报道。本项目就是以此为切入点，采用激素加内毒素建立兔SANFH模型，并以破瘀通络法和补肾通络法组方的中药辨证进行干预，通过组织学、原位末端标记法(TUNEL)和分子生物学等技术，检测和观察股骨头坏死过程中血管形成、新骨形成,CD34、CYR61、CTGF、VEGF表达和VEGFmRNA表达与细胞凋亡和bcl-2表达的变化、相互关系以及中药的影响，目的在于从血管形成与细胞凋亡相关角度，进一步研究SANFH的发病机制，探讨中药治疗SANFH的作用机理，为中药治疗股骨头坏死和激素引起的骨疾病提供依据。

激素性股骨头坏死是临床常见病，难治病。前期研究表明激素诱导的股骨头缺血性坏死是缺血和细胞凋亡共同作用的结果，促进股骨头坏死区域的血管重建及再生有利于坏死的修复。但是血管的重建再生与细胞凋亡是否相关以及调控血管形成的相关因子的研究未见报道。. 本研究采用内毒素联合激素建立典型的兔股骨头坏死模型通过建立兔股骨头缺血坏死模型，后根据血脂的水平辩证为血瘀型和肾虚型，以破瘀通络法及补肾通络法组方的中药辩证干预。分别在造模后4周、8周和12周取出标本通过组织学、免疫组化、原位末端标记（TUNEL）及逆转录-聚合酶链反应（RT-PCR）等观察股骨头中坏死区域结构形态的改变，股骨头血管内皮细胞标记抗原(CD34)、富含半胱氨酸61（CYR61）、结蹄组织生长因子（CTGF）、血管内皮生长因子（VEGF）和bcl-2表达的变化，细胞凋亡情况以及VEGF mRNA表达的变化等来进一步研究激素性股骨头坏死的发生机制以及中药防治激素性股骨头坏死的作用机理。. 实验结果表明：1、骨蚀宁组和健骨生两组空骨陷窝数百分比明显降低。2、骨蚀宁组与健骨生两组股骨头CD34、CYR61、CTGF和血VEGF表达偏高。3、骨蚀宁组与健骨生组细胞凋亡率偏低。4、骨蚀宁组与健骨生两组bcl-2表达偏高。4、骨蚀宁组和健骨生两组VEGF mRNA表达偏高。. 总之，辩证运用中药骨蚀宁胶囊Ⅰ号和Ⅱ防治激素性股骨头坏死的实验研究取得了显著的治疗结果，其作用是多途径的，不仅能调控相关因子表达促进血管的再生，又能抑制细胞凋亡，促进坏死组织结构的重建和修复。本课题的研究必将为中药治疗股骨头坏死和激素引起的骨疾病提供依据。