骨折愈合过程中生长因子表达与中药阿胶复方的干预效应

背景:阿胶强骨口服液可有效治疗骨折,但其具体的药理学机制仍有待研究。在骨折愈合中,血管内皮生长因子和成纤维细胞生长因子2是促进血管内生以及骨的合成代谢的重要耦联因子。目的:观察阿胶强骨口服液对SD大鼠胫骨骨折愈合过程中骨痂中血管内皮生长因子和成纤维细胞生长因子2表达的影响,探讨阿胶强骨口服液治疗骨折的疗效机制。设计:完全随机对照实验。单位:华中科技大学同济医学院附属协和医院骨伤科研究室。材料:选用90只雌性3月龄SD大鼠,体质量(368±40)g,阿胶强骨口服液(主要成分为阿胶,新疆华世丹药业股份有限公司生产);阳性对照药接骨七厘片(主要成分为当归、乳香、没药、大黄、血竭、骨碎补、自然铜,珠海金沙(湖南)制药有限公司生产)。方法:实验于2005-07/12在华中科技大学同济医学院中西结合骨代谢实验室(省级实验室)完成,采用三点弯曲方法造成大鼠右胫骨中段闭合性骨折后,采用完全随机法分为3组:阿胶强骨口服液组(n=30):按人鼠表面积比率换算等效计量法计算后,每只每次给予阿胶强骨口服液2mL灌胃给药,2次/d;接骨七厘片组(n=30):用蒸馏水配制成225g/L灌胃,2次/d;生理盐水组(n=30):给予同等容量,同等频率的生理盐水灌胃。分别在实验第4,7,14,21,28天采用免疫组织化学方法检测大鼠右胫骨骨痂区血管内皮生长因子和成纤维细胞生长因子2表达的变化,计算平均吸光度值及阳性细胞数(5个视野细胞)。主要观察指标:各组大鼠右胫骨骨痂区血管内皮生长因子和成纤维细胞生长因子2表达(平均吸光度值及阳性细胞数)。结果:纳入大鼠90只均进入结果分析。①血管内皮生长因子表达检测结果:实验开始后4,7,14d,阿胶强骨口服液组大鼠及接骨七厘片组平均吸光度值均高于生理盐水组(P<0.05～0.01),阳性细胞数变化幅度与平均吸光度值一致。②成纤维细胞生长因子2表达检测结果:实验开始后4,7d,阿胶强骨口服液组及骨七厘片组成纤维细胞生长因子2平均吸光度值,均高于生理盐水组(P<0.05～0.01),阳性细胞数变化幅度与平均吸光度值一致。结论:阿胶强骨口服液在骨愈合早中期可通过调节血管内皮生长因子和成纤维细胞生长因子2的表达,促进前成骨细胞和成软骨细胞的有丝分裂、血管内生以及骨的合成代谢达到促骨折愈合的作用。

Interventional effects of donkey-hide glue reinforcing bone oral solution on the expression of growth factors during fracture healing

BACKGROUND:Donkey-hide glue reinforcing bone oral solution (DGRBOS) is effective on preventing and treating fracture, but the mechanism of harmacology is still not clear. Vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) are important cytokines, which can promote blood vessel growth and osseous anabolism during fracture healing.OBJECTIVE: To investigate the effects of DGRBOS on expression of VEGF and FGF-2 in the process of fracture healing of SD rats' fracture of tibia in bony callus, and explore the mechanism of DGRBOS in the treatment of fracture.DESIGN: A completely randomized controlled study.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: Ninety female Sprague-Dawley (SD) rats, with a mean weight of (368±40) g, aged about 3 months, wereprovided by Center of Animal Experiment, Tongji Medical College, Huazhong University of Science and Technology.Experimental drug: DGRBOS was donated by Xinjiang Huashidan Pharmaceutical Co., Ltd. The qili linking bone pill,positive control drug, was purchased from Hunan Pharmaceutical Co., Ltd.METHODS: This experiment was carried out in the Laboratory for Bone Metabolism of Integration of Chinese and Western Medicine (Laboratory of Provincial Level), Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June to November 2005. Transverse midshaft fractures were produced on the right tibia in these rats using three-point bending technique, and 90 rats were randomly divided into three experimental groups:the DGRBOS group (n =30): according to medicine conversional method between human being and animal, 2 mL DGRBOS was fed to every mouse by intragastric administration at 2 vices/day; the qili linking bone pill group (positive control group, n =30): the pills dissolved in distilled water at 225 g/L, and consequently were administered intragastrically into mice at 2 mL/vice and 2 vices/day; normal saline group (negative control group, n =30): normal saline was administered intragastrically into mice at the coordinative capacity and same frequences. Selecting 4, 7, 14,21 and 28 days during the experiment, the immunohistochemistry method was adopted to detect the change of expression of VEGF and FGF-2 through computing value of mean optical degree (MOD) and number of the positive cells.