十味接骨药对家兔骨折骨细胞生长机理实验研究.docVIP

十味接骨药对家兔骨折骨细胞生长机理实验研究 　　【摘要】 目的 研究十味接骨药对家兔骨折骨细胞生长修复过程中骨痂中TGF-β?1,羟脯氨酸、钙、磷;血清中碱性磷酸酶、钙、磷的变化和作用机理。方法 实验用健康日本大白兔81只5~6月龄造成双侧前肢0.7 cm横断性骨缺损,保留尺骨完整,并分成三组,对照组和实验组,空白组不用药,实验组给十味接骨药,对照组给骨折挫伤散。2周、5周、8周分别取材,观察十味接骨药对骨折修复的影响。结果 实验组骨痂中TGF-β?1、HRP先升高后递减;血中骨痂中Ca、P峰值提前出现,血中ALP随骨生长逐渐升高。与对照组相比较统计学?*P 　　【关键词】 骨折愈合;骨细胞生长;十味接骨药 　　 　　Therapeutic Effect of SHI-WEI-JIE-GU-YAO on Experimental Fracture and Its Mechanism 　　 　　TIAN Qing-shan, SHUI Guo-xing,LI Kun,et al. 　　Jiamusi College, Heilongjiang University of Chinese Medicine, Jiamusi, Heilongjiang 154002,China 　　 　　【Abstract】 Objective To study the effect of SHI-WEI-JIE-GU-YAO in the healing process of fracture. Methods Eighty-one rabbits, aged 5-6 months, had been made experimental fractures at the middle shaft of the humerus bilaterally and divided into 3 groups randomly: the blank control group, fed with common forage; the experimental group, fed with SHI-WEI-JIE-GU-YAO additionally; the drug control group, fed with GU-ZHE-CUO-SHANG-SAN. The specimens were taken out 2, 5, and 8 weeks after operation. The contents of TGF-β?1、hydroxyproline、calcium、phosphorus in callus and alkaline phosphatase、calcium、phosphorus in serum were tested respectively. Results All the data showed that TGF-β?1, hydroxyproline in calluses of experimental group were raised firstly and descended afterwards, the peak value of calcium and phosphorus appeared in advance compared with other groups (P 　　1.6 观察指标与方法 　　1.6.1 血样分析 三组动物按三个时间段进行P-ALP、S-Ca、S-P分析,将兔放兔盒中固定,选耳后静脉清晰部位拔除局部绒毛,75%乙醇消毒,摩擦静脉使之扩张,用6号针头在耳缘静脉末稍刺破血管,待血渗出后用注射器抽取血液2 ml放不加抗凝剂试管中,用80-Z离心机离心制成血清,用奥林巴斯AU-2700全自动生化分析仪分析血清钙(Ca)、磷(P)、碱性磷酸酶(ALP)。 　　1.6.2 骨痂分析 羟脯氨酸(HRP)、钙(Ca)、磷(P)。将新鲜??痂用丙酮脱脂脱水12 h真空干燥称重,用10%三氯乙酸脱钙脱磷,用甲基百香酚蓝比色法测提取液中的钙;用孔雀绿直接显色法测提取液中的磷,剩余骨痂用蒸馏水冲洗,滤纸吸去多余的水分称重,用样本碱水解法测骨痂中的羟脯氨酸。具体做法:①骨痂标本丙酮脱脂脱水,用TH2-88-1型多用恒温台式振荡器振荡3 h后,静置12 h,使脱水完全取出样本置于真空干燥皿中,用SHB-Ⅲ循环水式多用真空泵真空干燥后,用MettierH54AR十万分之一分析电子天平称重,将干燥样本浸在3 ml,10%三氯乙酸中提取样本中的钙、磷24 h取出样本待用。按试剂盒说明方法测钙、磷;②用甲基百香酚蓝比色法测提取液中的钙(Ca)含量,空白管、标准管,