| 淫羊藿总黄酮对去势大鼠骨折原始骨痂发生的影响及其机制 |
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Received:January 18, 2017
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| 作者 | Author | 单位 | Unit | E-Mail |
| 史晓林 |
SHI Xiao-lin |
浙江中医药大学附属第二医院, 浙江 杭州 310017 |
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| 梁博程 |
LIANG Bo-cheng |
浙江中医药大学附属第二医院, 浙江 杭州 310017
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 吴鹏 |
WU Peng |
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 王博 |
WANG Bo |
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 施振宇 |
SHI Zhen-yu |
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 孔令成 |
KONG Ling-cheng |
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 姚建亮 |
YAO Jian-liang |
浙江中医药大学第二临床医学院, 浙江 杭州 310053 |
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| 黄俊俊 |
HUANG Jun-jun |
浙江中医药大学附属第二医院, 浙江 杭州 310017 |
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| 李春雯 |
LI Chun-wen |
浙江中医药大学基础医学院, 浙江 杭州 310053 |
College of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
xlshi-2002@163.com |
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| 期刊信息：《中国骨伤》2017年30卷，第8期，第743-750页 |
| DOI：10.3969/j.issn.1003-0034.2017.08.013 |
| 基金项目:国家自然科学基金面上项目（编号：81373878）；高校博士点基金联合项目（博导类）（编号：20133322110005）；浙江省自然科学基金一般项目（编号：LY14H270003） |
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目的：探讨淫羊藿总黄酮对去势大鼠骨质疏松性骨折愈合中骨痂形成的影响。
方法：选取40只6~8周龄，体重209~246 g的雌性SD大鼠，采用钢锯截断加克氏针髓内固定法，建立去势大鼠股骨中段骨质疏松性骨折模型。按随机数字表法随机分为淫羊藿总黄酮组和对照组，每组20只。淫羊藿总黄酮组给予淫羊藿总黄酮150 mg·kg-1·d-1，对照组给予等量生理盐水口服。在干预6周后，使用双能X线骨密度仪检测股骨BMD，使用Micro CT系统测量骨痂的骨体积、骨体积分数、骨小梁厚度、骨小梁数量及骨小梁分离度等参数。采用HE染色技术观察两组骨痂愈合进程中的骨痂组织形态学差异，采用免疫组织化学染色技术检测骨痂处酪蛋白激酶2相互作用蛋白1（casein kinase 2 interacting protein 1，CKIP-1）的表达量。采用RT PCR技术检测两组骨痂样本中骨特异性转录因子2（runt-related transcription factor 2，Runx2）表达量，采用3点弯曲力学方法测量骨痂处最大载荷值。
结果：干预6周后，淫羊藿总黄酮组骨密度为（129.4±3.1） mg/cm2，与对照组的（117.3±3.3） mg/cm2比较差异有统计学意义（t=-4.628 8，P<0.001）；Micro CT扫描结果显示，淫羊藿总黄酮组和对照组在骨痂骨小梁数量、骨痂体积、骨痂体积/总体积、骨小梁分离度和骨小梁厚度方面比较差异有统计学意义（P<0.05）。200倍和400倍光镜下骨痂HE染色切片，见骨小梁微结构参数与骨痂Micro CT扫描结果趋势一致，同时发现淫羊藿总黄酮组软骨细胞骨化水平明显高于对照组。骨痂中Runx2因子RT PCR检测结果显示，两组的Runx2 mRNA相对表达量差异有统计学意义（P<0.001）。400倍镜下观察骨痂中CKIP-1蛋白免疫组化检测结果显示，淫羊藿总黄酮组和对照组CKIP-1蛋白阳性表达数分别为9.30±1.16，40.50±1.08，差异有统计学意义（t=-62.26，P<0.001）。骨痂生物力学结果显示，淫羊藿总黄酮组和对照组最大载荷分别为（98.37±9.64）、（68.45±6.07） N，差异有统计学意义（t=6.433 4，P<0.001）。
结论：淫羊藿总黄酮可以促进去势大鼠骨质疏松性骨折愈合过程中成骨细胞骨形成能力和软骨骨化能力，促进原始骨痂的形成，增强骨折愈合强度，而其促进骨愈合和增强骨折愈合强度可能是通过调节CKIP-1蛋白来实现的。 |
| [关键词]:淫羊藿总黄酮 原始骨痂 骨形成 软骨内成骨 大鼠 |
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| Effect and mechanism of total flavone of epimedium on primary callus formation in ovariectomized rats with fractures |
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Abstract:
Objective: To explore the effects and related mechanisms of total flavone of epimedium treatment(TFE)on primary callus for mation in ovariectomized rats.
Methods: Forty male SD rats weighted from 209 to 246 g and aged 6 to 8 weeks were selected. Six weeks after ovariectomy a femur fracture model with middiaphyseal segment fracture was established,estimated and randomly divided into TFE group (150 mg·kg-1·d-1) and control group(received saline). HE staining was used to evaluate the morphologic difference of primary callus during the bone callus healing between these two groups. The relative expression of Runt-related transcription factor 2(Runx2) mRNA in the callus was identified by real time polymerase chain reaction. Immunohistochemical technique was used to observe the Casein kinase 2 interacting protein 1(CKIP-1) protein level in the callus of the two groups. Maximum fracture load was tested by three point bend test.
Results: The BMD,primary callus volume,trabecular member(Tb.N) and trabecular thickness(Tb.Th) were higher in TFE group than that in control group(P<0.001). The Tb.N and Tb.Th of primary callus were higher in TFE group than control group (P=0.001). The volume and bone volume/tissue volume of primary callus were in TFE group than control group(P<0.01). The trabecular separation(Tb.Sp) of primary callus were in control group higher than TFE group(P<0.01). The HE staining of the 6 week slices showed that the degree of cartilage ossification in callus of the TFE group was significantly higher than that in control group under high magnification. Real time PCR revealed that the comparative expression of Runx2 mRNA in control group was higher than that in TFE group(P<0.001);the positive number of CKIP-1 was less in TFE group than that in control group (P<0.001). TFE could increase the maximum load of the primary callus (P<0.001).
Conclusion: TFE can promote the cartiage ossification of callus in ovariectomized rats,enhancing the bone strength and bone quality in the process of fracture healing via the CKIP-1/Runx2 pathway. |
| KEYWORDS:Total flavone of epimedium Primary callus Bone formation Endochondral ossification Rats |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 史晓林,梁博程,吴鹏,王博,施振宇,孔令成,姚建亮,黄俊俊,李春雯.淫羊藿总黄酮对去势大鼠骨折原始骨痂发生的影响及其机制[J].中国骨伤,2017,30(8):743~750 |
| 英文格式: | SHI Xiao-lin,LIANG Bo-cheng,WU Peng,WANG Bo,SHI Zhen-yu,KONG Ling-cheng,YAO Jian-liang,HUANG Jun-jun,LI Chun-wen.Effect and mechanism of total flavone of epimedium on primary callus formation in ovariectomized rats with fractures[J].zhongguo gu shang / China J Orthop Trauma ,2017,30(8):743~750 |
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