西伯利亚接骨木树皮提取物促进大鼠骨折愈合的机制研究

The mechanism of bark extract of Sambucus sibirica Nakai promoting fracture healing in rats

目的初步探讨新疆西伯利亚接骨木树皮不同提取物促进大鼠骨折愈合的作用机制。方法建立大鼠股骨开放性骨折模型,将建模成功的96只大鼠随即分为8组:模型对照组(蒸馏水,0.2 mL/kg体质量),阳性药物组(中华跌打丸0.54 g/kg体质量),水提取物高、中、低(0.340、0.170、0.085 g/kg体质量)剂量组,醇提取物高、中、低(0.314、0.157、0.078 g/kg体质量)剂量组,另取12只未建模的大鼠为空白对照组(蒸馏水,0.2 mL/kg体质量),分别在药物干预14、28、42 d时取血样和股骨样本,采用酶联免疫法测定大鼠血清中碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、骨钙素(BGP);骨保护素(OPG)、Ⅰ型胶原羧基端肽(PICP)水平,采用苏木精-伊红(HE)染色观察大鼠股骨骨组织形态学变化。结果与模型组比较,西伯利亚接骨木醇提高、中剂量组,水提高剂量组和阳性药物组大鼠血清ALP、ACP、BGP、PICP水平增高,OPG水平降低,差异有统计学意义(P<0.05)。与阳性药物组比较,14 d时西伯利亚接骨木醇提高剂量组ALP、ACP、BGP、PICP水平增高,28 d和42 d时西伯利亚接骨木醇提高剂量组ACP水平增高,差异有统计学意义(P<0.05)。与阳性药物组,14、28和42 d时西伯利亚接骨木醇提中、低剂量组,水提中、低剂量组大鼠血清OPG水平均增高,差异有统计学意义(P<0.01)。光镜观察显示:接骨木醇提、水提高剂量组和阳性药物组新生骨小梁增多,可见成骨细胞和破骨细胞。接骨木醇提和水提中、低剂量组及模型组骨折端骨外膜可见少量新生骨小梁,类骨质较少。结论新疆西伯利亚接骨木和跌打丸治疗的大鼠骨代谢情况更佳,促进成骨细胞和破骨细胞的合成,增强成骨细胞和破骨细胞的活性,从而促进骨折愈合。

Objective To explore the mechanism of different bark extracts of Sambucus sibirica Nakai in Xinjiang promoting fracture healing in rats.Methods96 rats were randomly divided into 8 groups:model control group(distilled water,0.2 mL/kg body mass),positive drug group(0.54 g/kg body mass),water extract high,medium and low(0.340 g/kg body mass,0.170 g/kg body mass and 0.085 g/kg body mass),alcohol extract high,medium and low(0.314 g/kg body mass,0.157 g/kg body mass,0.078 g/kg body mass),another 12 normal rats were taken as the blank control group(distilled water,0.2 mL/kg body mass).The open fracture model of femur was established.After drug intervention,the blood samples and femur samples were taken at 14 days,28 days and 42 days.The level of alkaline phosphatase(ALP),acid phosphatase(ACP),osteocalcin(BGP),osteoprotegerin(OPG)and propeptide carboxy-terminal procollagentypeⅠ(PICP)was measured by enzyme-linked immunosorbent assay(ELISA),and the histomorphologyof bone was observed by hematoxylin eosin(HE)staining.ResultsCompared with the model group,the level of serum ALP,ACP,BGP and PICP was increased and the level of OPG was decreased in the alcohol extract high and medium dose group and water extract high dose group of S.sibirica Nakai and positive drug group(P<0.05).Compared with the positive drug group,the level of ALP,ACP,BGP and PICP in the extract high dose group was increased at day 14,and the level of ACP in the extract high dose group was increased at day 28 and day 42,with statistically significant differences(P<0.05).The serum OPG level in the middle and low dose groups was significantly higher than those in the positive drug group(P<0.01).The number of new bone trabeculae was increased in the alcohol extract and water extract high dose group and the positive drug group.A small amount of new bone trabeculae and osteoid were found in the periosteum of the fracture end in the middle and low dose groups and the model group.ConclusionThe extract of S.sibirica Nakai and Dieda pill have better effects on the bone metabolism in rats,promote the synthesis of osteoblasts and osteoclasts,enhance the activity of osteoblasts and osteoclasts,and thus promote fracture healing.