补肾活血方对沙鼠增龄过程中腰椎软骨终板钙化干预作用的初步研究
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作者Author单位AddressE-Mail
朱立国 ZHU Li-guo 中国中医科学院脊柱二科, 北京 100102 Department of Pharmacology, Beijing 100102, China  
张平 ZHANG Ping 中国中医科学院望京医院病理科, 北京 100102 Wangjing Hospital of China Academy of Traditional Chinese Medical Scinences, Beijing 100102, China pinglele@sina.com 
宋庆慧 SONG Qing-hui 中国中医科学院望京医院药理实验室, 北京 100102  
张万强 ZHANG Wan-qiang 中国中医科学院望京医院药理实验室, 北京 100102  
朱宏伟 ZHU Hong-wei 中国中医科学院望京医院病理科, 北京 100102 Wangjing Hospital of China Academy of Traditional Chinese Medical Scinences, Beijing 100102, China  
杨丽平 YANG Li-ping 中国中医科学院望京医院药理实验室, 北京 100102  
朱岩 ZHU Yan 中国中医科学院望京医院病理科, 北京 100102 Wangjing Hospital of China Academy of Traditional Chinese Medical Scinences, Beijing 100102, China  
王源 WANG Yuan 中国中医科学院望京医院药理实验室, 北京 100102  
李玲慧 LI Ling-hui 中国中医科学院骨伤综合科, 北京 100102  
期刊信息:《中国骨伤》2017年,第30卷,第10期,第926-932页
DOI:10.3969/j.issn.1003-0034.2017.10.010
基金项目:国家中医药管理局国家中医临床研究基地业务建设科研专项课题(编号:JDZX2015275);国家自然科学基金面上项目(编号:81674005);北京市自然科学基金青年项目(编号:7164313);中国博士后科学基金面上项目(编号:2016M591364)。
中文摘要:

目的:探讨补肾活血方对腰椎软骨终板钙化的干预作用。

方法:选取30只2月龄和7月龄健康雄性长爪沙鼠,分别喂养至2月龄(50~60 g)和12月龄(60~80 g)建立老龄沙鼠模型,采用随机数字表方法分为5组:正常组(n=6),模型组(n=6,给予生理盐水4 ml/kg,灌胃30 d),补肾活血方低剂量组(n=6,给予补肾活血方1.9×10-3 ml/g灌胃30 d),补肾活血方中剂量组(n=6,给予补肾活血方3.8×10-3 ml/g灌胃30 d),补肾活血方高剂量组(n=6,给予补肾活血方7.6×10-3 ml/g灌胃30 d),干预组从7月龄开始连续给药1.36 g,30 d.正常组2月龄及其他组12个月龄时处死动物,取腰椎间盘组织,HE染色体视法分析腰椎体软骨终板组织形态学、血管芽面积、非钙化/钙化层比值,兔单克隆抗体免疫组化染色方法测定软骨终板X型胶原、BMPs的表达。

结果:软骨终板血管芽相对面积测量显示,与模型组相比,补肾活血方中剂量组,正常组增高(P<0.05),大、小剂量组虽各有不同程度的增加,但差异均无统计学意义(P>0.05);软骨终板非钙化层/钙化层厚度比值测量显示,与模型组相比,补肾活血方中剂量、正常组增加,差异具有统计学意义(P<0.05),而补肾活血方大、小剂量组虽各有不同程度的增加,但差异均无统计学意义(P>0.05).免疫组化染色软骨终板X型胶原的表达与模型组相比,正常组、补肾活血方低、中、高剂量组均降低,差异具有统计学意义(P<0.01);BMPs的表达,与模型组相比,正常组、补肾活血方中剂量组升高,差异具有统计学意义(P<0.01),大、小剂量组虽各有不同程度的升高,但差异均无统计学意义(P>0.05).

结论:补肾活血方中剂量可延缓增龄过程中椎间盘软骨终板钙化,提示可作为早期椎间盘退变的预防用药。
【关键词】补肾活血方  椎间盘退变  软骨终板钙化  沙鼠
 
Preliminary study of intervention in effect of Bushen Huoxue recipe on calcification of lumbar vertebra cartilage endplate of the aging gerbils
ABSTRACT  

Objective:To study the effect of Bushen Huoxue decoction on calcification of cartilage endplate in lumbar vertebrae.

Methods:Six healthy male gerbils with 2-month-old were selected as normal control group,and 24 7-month-old healthy male gerbils were fed to 12-month-old to establish the aged gerbil model. Thirty gerbils were randomly divided into five groups as follow:the normal control group (n=6),model group (n=6,normal saline 4 ml/kg,intragastric 30 d),Bushen Huoxue low dose group(n=6,1.9×10-3 ml/g given Bushen Huoxue recipe orally,30 d),Bushen Huoxue middle dose group(n=6,3.8×10-3 ml/g given Bushen Huoxue recipe orally,30 d),Bushen Huoxue high dose group(n=6,7.6×10-3 ml/g given Bushen Huoxue recipe orally,30 d),the intervention group administered for 1.36 g from 7-month-old age,30 d. The animals were sacrificed at the age of 2 months in the normal control group and 12 months of age in the other groups. The morphology of the lumbar vertebral cartilage endplate,the area of vascular bud,the ratio of non-calcified/calcified layer were analysis by HE chromosome visual method. The expression of type X collagen and BMPs in cartilage endplates were detected by rabbit monoclonal immunohistochemical staining.

Results:The relative area of the vascular buds cartilage endplate measurements showed that compared with the model group,middle dose group and normal control group increased (P<0.05),high and low dose groups all had different degrees of increase,but no statistical significance(P>0.05). The ratio of cartilage endplate thickness of non-calcified/calcified showed that compared with the model group,Bushen Huoxue middle dose,normal control group increased,with statistical significance(P<0.05),and high and low dose groups all had different degrees of increase,but there were no statistical significance(P>0.05). Compared with the model group,the expression of type X collagen in the cartilage endplate of the normal group,the Bushen Huoxue low,middle and high dose groups decreased,and had statistical significance(P<0.01);compared with the model group,the expression of BMPs in the normal group,Bushen Huoxue middle dose group increased,with statistically significant(P<0.01),while the high and low dose groups increased in different degrees,but there was no statistical significance(P>0.05).

Conclusion:Bushen Huoxue prescription can delay the calcification of cartilage endplate in the process of aging,suggesting that it can be used as a preventive medicine for early disc degeneration.
KEY WORDS  Bushen Huoxue recipe  Intervertebral disc degeneration  Cartilage endplate calcification  Gerbils
 
引用本文,请按以下格式著录参考文献:
中文格式:朱立国,张平,宋庆慧,张万强,朱宏伟,杨丽平,朱岩,王源,李玲慧.补肾活血方对沙鼠增龄过程中腰椎软骨终板钙化干预作用的初步研究[J].中国骨伤,2017,30(10):926~932
英文格式:ZHU Li-guo,ZHANG Ping,SONG Qing-hui,ZHANG Wan-qiang,ZHU Hong-wei,YANG Li-ping,ZHU Yan,WANG Yuan,LI Ling-hui.Preliminary study of intervention in effect of Bushen Huoxue recipe on calcification of lumbar vertebra cartilage endplate of the aging gerbils[J].zhongguo gu shang / China J Orthop Trauma ,2017,30(10):926~932
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