天津医药

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Wnt/β-catenin信号通路与骨关节炎发病机制的研究

王超华1,邱明才2,刘军3,李凤翱1,4,卢飚1,4,任凯晶5,张子博1,4,朱梅1   

  1. 1. 天津医科大学总医院内分泌科
    2. 天津医科大学总医院
    3. 天津市人民医院
    4.
    5. 天津医院关节外科
  • 收稿日期:2010-12-02 修回日期:2011-02-17 出版日期:2011-06-15 发布日期:2011-06-15
  • 通讯作者: 朱梅

Wnt/β-catenin signaling in the pathogenesis of osteoarthritis

  • Received:2010-12-02 Revised:2011-02-17 Published:2011-06-15 Online:2011-06-15

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摘要： 摘要 目的 观察行膝关节置换术的骨关节炎(osteoarthritis，OA)患者软骨组织Wnt/β-catenin 信号传导通路相关蛋白及基因的表达情况。方法 关节软骨组织经石蜡包埋切片后分别行HE、番红O/固绿和阿辛蓝/橙黄G三种染色。根据改良的Mankin关节软骨病理评分标准进行分级评定，确定关节软骨的损伤程度。应用免疫组化的方法检测软骨细胞中β-catenin及MMP-13蛋白的表达情况。提取行膝关节置换术OA患者的关节软骨总RNA，应用Real Time RT-PCR的方法，选用B2M作为内参，检测BMP-2，COL-10及MMP-13基因的表达情况，并比较各病变组上述基因的表达差异。结果 1. 改良的Mankin病理评分结果：对照组：8例，均正常。实验组：轻度病变组15例，中度病变组27例。2.β-catenin免疫组化结果：对照组全部为阴性。实验组：轻度病变组9例阳性，阳性率60.0%；中度病变组7例阳性，阳性率25.9%。MMP-13免疫组化结果：对照组1例阳性，阳性率12.5%。实验组：轻度病变组14例阳性，阳性率93.3%；中度病变组21例阳性，阳性率77.8%。3.Real Time RT-PCR结果：轻度病变组BMP-2的表达水平为中度病变组的3.01倍，COL-10为3.89倍，MMP-13为4.08倍，均具有统计学差异（P<0.05）。结论 1.OA患者关节软骨细胞中β-catenin及MMP-13蛋白的表达水平增加，后者增加更为明显。2.OA患者关节软骨细胞分化的标志性基因BMP-2、COL-10、MMP-13的表达水平显著增加。3.β-catenin在关节软骨细胞中高表达可能是关节软骨破坏并导致OA发展的重要原因。

关键词: 骨关节炎, 软骨细胞, β-catenin, 骨形态发生蛋白-2, X型胶原, 基质金属蛋白酶-13

Abstract: Abstract Objectives The expression of proteins and genes of Wnt/β-catenin signaling in the articular cartilage of human OA samples from patients undergoing total knee arthroplasty.Methods Knee joint samples were dissected and embeded in paraffin. The tissue sections were stained with HE , Safranin O / Fast Green (SO/FG) and Alcian blue / orange G(AB/OG),and graded using a modified version of the Mankin scale. Immunohistochemistry staining of β-catenin and MMP-13 were performed in all samples. Total RNA extracted from articular cartilage tissue was prepared using Trizol according to the manufacture’s protocol. Real time RT-PCR was performed to test BMP-2, COL-10 and MMP-13 expression. Results 1.All samples were studied and the distribution of averaged grades allowed for stratification of human samples into three groups :control ,mild OA and moderate OA. 2.Immunohistochemistry of all samples showed a significant up-regulation of β-catenin and MMP-13 in both the mild and moderate groups compared with the normal control.3.Expression of chondrocyte marker genes such as BMP-2, COL-10 and MMP-13 were significantly increased in moderate OA group. Conclusions 1.The protein level of β-catenin and MMP-13 was increased in OA patients.2.Expression of chondrocyte maturation marker genes, such as BMP-2, COL-10, and MMP-13 were significantly increased in articular cartilage of OA patients. 3.Overexpression of β-catenin in articular chondrocytes leads to premature chondrocytes differentiation and the development of OA. These results establish a strong association between human OA and β-catenin expression.

Key words: osteoarthritis, articular chondrocytes, β-catenin, bone morphogenetic protein -2, type X collagen, matrix metalloproteinase-13