Activation of Indian hedgehog promotes chondrocyte hypertrophy and upregulation of MMP-13 in human osteoarthritic cartilage.
Activation of Indian hedgehog promotes chondrocyte hypertrophy and upregulation of MMP-13 in human osteoarthritic cartilage.
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DOI：
10.1016/j.joca.2012.03.010
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发表时间：
2012-07
影响因子：
7
通讯作者：
Wei, L.
中科院分区：
文献类型：
--
作者：
Wei, F.;Zhou, J.;Wei, X.;Zhang, J.;Fleming, B. C.;Terek, R.;Pei, M.;Chen, Q.;Liu, T.;Wei, L.
本研究的目的是1）确定骨关节炎（OA）和Ihh表达之间的相关性，和2）建立Ihh对人OA软骨中软骨细胞肥大和MMP-13的标志物表达的影响。在全膝关节置换术期间获得OA软骨和滑液样本。正常软骨样品从关节内肿瘤切除获得，正常滑液样品从健康志愿者和接受前交叉韧带重建的患者的对侧未受伤的膝关节获得。使用Mankin评分对OA进行分级。免疫印迹法检测滑膜液中Ihh的表达。采用真实的实时荧光定量PCR检测Ihh、X型胶原和MMP-13 mRNA的表达。免疫组化法检测软骨组织中X型胶原和MMP-13的蛋白表达。使用图像分析测量软骨细胞大小。与正常对照样本相比，OA软骨中Ihh表达增加2.6倍，OA滑液中Ihh表达增加37%。Ihh的表达增加与OA的严重程度和软骨细胞肥大的标志物X型胶原和MMP-13的表达以及软骨细胞大小相关。随着OA严重程度的增加，软骨细胞更加球形。Mankin评分与细胞大小（r2= 0.80）和Ihh强度（r2 = 0.89）之间存在显著相关性。外源性Ihh诱导培养的软骨细胞中X型胶原的表达增加6.8倍，MMP-13 mRNA的表达增加2.8倍。相反，通过siRNA和Hh抑制剂环巴胺敲低Ihh具有相反的效果。Ihh表达与OA进展和软骨细胞形态学变化以及与OA软骨中观察到的软骨细胞肥大和软骨降解一致的基因表达相关。因此，Ihh可能是预防OA进展的潜在治疗靶点。
The objectives of this study were to 1) determine the correlation between osteoarthritis (OA) and Ihh expression, and 2) establish the effects of Ihh on expression of markers of chondrocyte hypertrophy and MMP-13 in human OA cartilage. OA cartilage and synovial fluid samples were obtained during total knee arthroplasty. Normal cartilage samples were obtained from intra-articular tumor resections, and normal synovial fluid samples were obtained from healthy volunteers and the contralateral uninjured knee of patients undergoing anterior cruciate ligament reconstruction. OA was graded using the Mankin score. Expression of Ihh in synovial fluid was determined by western blot. Ihh, type X collagen and MMP-13 mRNA were determined by real time PCR. Protein expression of type X collagen and MMP-13 in cartilage samples were analyzed with immunohistochemistry. Chondrocyte size was measured using image analysis. Ihh expression was increased 2.6 fold in OA cartilage and 37% in OA synovial fluid when compared to normal control samples. Increased expression of Ihh was associated with the severity of OA and expression of markers of chondrocyte hypertrophy: type X collagen and MMP-13, and chondocyte size. Chondrocytes were more spherical with increasing severity of OA. There was a significant correlation between Mankin score and cell size (r2= 0.80) and Ihh intensity (r2 = 0.89). Exogenous Ihh induced a 6.8 fold increase of type X collagen and 2.8 fold increase of MMP-13 mRNA expression in cultured chondrocytes. Conversely, knockdown of Ihh by siRNA and Hh inhibitor Cyclopamine had the opposite effect. Ihh expression correlates with OA progression and changes in chondrocyte morphology and gene expression consistent with chondrocyte hypertrophy and cartilage degradation seen in OA cartilage. Thus, Ihh may be a potential therapeutic target to prevent OA progression.
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