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活血通络方加不同引经药对股骨头坏死兔成骨相关因子的影响
Hits: 2321   Download times: 1599   Received:June 07, 2012    
作者Author单位UnitE-Mail
王荣田 WANG Rong-tian 中国中医科学院望京医院, 北京 100102 Wangjing Hospital of China Academy of Chinese Medical Sciences, Beijing 100102, China  
孔祥英 KONG Xiang-ying 中国中医科学院中药研究所, 北京 100700 Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China  
田能 TIAN Neng 中国中医科学院中药研究所, 北京 100700 Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China  
林娜 LIN Na 中国中医科学院中药研究所, 北京 100700 Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China linna888@163.com 
陈卫衡 CHEN Wei-heng 中国中医科学院望京医院, 北京 100102 Wangjing Hospital of China Academy of Chinese Medical Sciences, Beijing 100102, China drchenweiheng@163.com 
期刊信息:《中国骨伤》2013年26卷,第7期,第591-596页
DOI:10.3969/j.issn.1003-0034.2013.07.015
基金项目:国家自然科学基金面上资助项目(编号:30672770)


目的: 观察活血通络方配伍不同引经药对股骨头坏死兔血清骨钙素(BGP)、骨形态发生蛋白-2(BPM-2)含量及股骨头BMP-2 mRNA表达的影响,探讨其防治股骨头坏死的作用机制。

方法: 健康雄性日本大耳白兔98只,体重2.2~2.8 kg.84只兔经液氮冷冻左侧股骨头法建立股骨头坏死模型后,随机分为6组,每组14只:模型组,活血通络方组,以及活血通络方分别加牛膝、独活、细辛、桔梗组(分别简称活骨、牛膝、独活、细辛、桔梗组).另外14只设为伪手术组。造模后药物组分别给予相应的中药,模型和伪手术组给予生理盐水灌胃,所有动物皮下注射重组人粒细胞集落刺激因子(30 μg·kg-1·d-1,连续7 d).实验2、4周时分别取材,放射免疫法和酶联免疫法分别检测血清BGP和BMP-2含量,左侧股骨头行苏木精-伊红染色观察组织病理学改变,原位杂交法检测BMP-2 mRNA表达。

结果: 与伪手术组相比,模型组股骨头空骨陷窝率显着增多,2周时血清BGP升高,4周时血清中BMP-2含量及股骨头BMP-2 mRNA 降低。与模型组相比,活骨组空骨陷窝率降低,2、4周时血清BGP、BMP-2含量及股骨头BMP-2 mRNA的表达均升高。与活骨组比较:牛膝组4周时空骨陷窝率显着减少,血清BGP、BMP-2及股骨头BMP-2 mRNA升高;桔梗组空骨陷窝有降低趋势,且2、4时周血清BGP降低,2周时血清BMP-2降低,4周时股骨头BMP-2 mRNA降低;独活组2周时血清BMP-2含量和股骨头BMP-2 mRNA的表达升高;细辛组各项指标未见明显改变。

结论: 引经药牛膝可以进一步提高活血通络方上调血清BGP、BMP-2含量和股骨头内BMP-2 mRNA表达的作用,这可能是其促进骨成有效防治股骨头坏死的机制之一。
[关键词]:股骨头坏死  骨钙素  骨形态发生蛋白受体,Ⅱ型  中草药
 
Effects of promoting blood circulation to remove meridian obstruction combined with medicinal guides on related osteogenesis factors in rabbits with femoral head necrosis
Abstract:

Objective: To observe effects of blood circulation promoting compounds combined with medicinal guides on content of bone glaprotein(BGP),bone morphogenetic protein-2(BPM-2) and expression of BMP-2 mRNA in rabbits with femoral head necrosis,and explore its mechanism.

Methods: Ninety-eight healthy Spragur-Dawley male rabbits were collected and weighted 2.2 to 2.8 kg. Eighty-four rabbits were built femoral head necrosis model by freezing left femoral head in liquid nitrogen,then randomly divided into 6 groups,14 in each group. The 6 groups included model group,promoting blood circulation to remove meridian obstruction group,promoting blood circulation to remove meridian obstruction combined with achyranthes bidentata group,radix angelicae pubescentis,asarum group,and platycodon grandiflorum group,other 14 rabbits were sham operation group. While drug groups were administrated corresponding Chinese herb after molding,model group and shamp operation group were given saline. Recombinant human granulocyte-colony stimulating factor (30 μg·kg-1·d-1)were injected into all rabbits for 7 days. Samples were taken on the second and fourth week,the content of BGP and BMP-2 were detected by enzyme-linked immunosorbent assay(ELSA) and radioimmunoassay(RIA),histopathological changes of left femoral head were observed by Hematoxylin and Eeosin staining(HE),and expression of BMP-2 mRNA were tested by fluorescence in situ hybridization.

Results: Compared with sham operation group,the rate of empty lacunae femoral head were obviously increased in model group,and the content of BGP were increased on the second week,and BMP-2 and BMP-2 mRNA were decreased on the fourth week. Compared with model group,the content of BGP,BMP-2 and BMP-2 mRNA were higher both of the second and fourth week in promoting blood circulation to remove meridian obstruction group. The rate of empty lacunae femoral head were lower in achyranthes bidentata group,BGP,BMP-2 and BMP-2 mRNA were higher on the fourth week. The rate of empty lacunae femoral head were lower in platycodon grandiflorum group,and BGP were decreased on the second and fourth week,BMP-2 were lower on the second week,while BMP-2 mRNA were decreased on the fourth week;the content of BMP-2 and BMP-2 mRNA were increased in radix angelicae pubescentis group on the second week; while there was no change in asarum group.

Conclusion: Radix angelicae pubescentis can increase the content of BGP,BMP-2 and expression of BMP-2 mRNA,which is an effective mechanism of preventing femoral head necrosis.
KEYWORDS:Femur head necrosis  Osteocalcin  Bone morphogenetic protein receptors,typeⅡ  Drugs,Chinese herbal
 
引用本文,请按以下格式著录参考文献:
中文格式:王荣田,孔祥英,田能,林娜,陈卫衡.活血通络方加不同引经药对股骨头坏死兔成骨相关因子的影响[J].中国骨伤,2013,26(7):591~596
英文格式:WANG Rong-tian,KONG Xiang-ying,TIAN Neng,LIN Na,CHEN Wei-heng.Effects of promoting blood circulation to remove meridian obstruction combined with medicinal guides on related osteogenesis factors in rabbits with femoral head necrosis[J].zhongguo gu shang / China J Orthop Trauma ,2013,26(7):591~596
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