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刺猬信号在大鼠颞下颌关节软骨中的表达及其与佐剂诱导性骨关节炎的关系。

Hedgehog signal expression in articular cartilage of rat temporomandibular joint and association with adjuvant-induced osteoarthritis.

作者信息

Xu Ting, Xu Gaoli, Gu Zhiyuan, Wu Huiling

机构信息

Department of Stomatology, First Affiliated Hospital, Zhejiang University, Hangzhou, China.

School of Stomatology, Zhejiang University, Hangzhou, China.

出版信息

J Oral Pathol Med. 2017 Apr;46(4):284-291. doi: 10.1111/jop.12497. Epub 2016 Sep 20.

Abstract

OBJECTIVE

The aim of this study was to investigate the changes in hedgehog (Hh) expression and its possible effects on cartilage degeneration in adjuvant-induced temporomandibular joint osteoarthritis (TMJOA) of rats.

METHODS

Forty-eight male Sprague-Dawley rats were randomly divided into experimental osteoarthritis (OA) and sham control groups. The bilateral TMJs of six rats from each group were harvested at three, seven, 14, and 28 days. Histological changes in condylar cartilage were assessed by hematoxylin and eosin, toluidine blue, and safranin O staining. The expression of Hh signal-related proteins including Indian hedgehog (Ihh), patched-1 (Ptch1), smoothened (Smo), glioma-associated oncogene homologue1 (Gli1) in cartilage was assessed by immunohistochemistry and Western blot. The protein expression of matrix metalloproteinase-13 (MMP-13), type X collagen, and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5) in cartilage was evaluated by Western blot.

RESULTS

The histological analysis showed marked cartilage degeneration in adjuvant-induced OA groups, including reduced cartilage cellular density, thinner and degraded cartilage, and decreased proteoglycan content in the extracellular matrix. Compared with matched control groups, the expression of Ihh, Ptch1, Smo, and Gli1 in the OA groups was higher in a time-dependent manner. The protein levels of MMP-13, type X collagen, and ADAMTS-5 were substantially increased in OA cartilage compared with those in matched control rats.

CONCLUSION

These results indicate that the activation of Ihh signaling may be correlated with pathological changes of condylar cartilage in adjuvant-induced TMJOA.

摘要

目的

本研究旨在探讨大鼠佐剂诱导性颞下颌关节骨关节炎(TMJOA)中刺猬信号通路(Hh)表达的变化及其对软骨退变的可能影响。

方法

48只雄性Sprague-Dawley大鼠随机分为实验性骨关节炎(OA)组和假手术对照组。每组6只大鼠在第3、7、14和28天处死,双侧颞下颌关节取材。采用苏木精-伊红染色、甲苯胺蓝染色和番红O染色评估髁突软骨的组织学变化。采用免疫组织化学和蛋白质印迹法评估软骨中刺猬信号相关蛋白,包括印度刺猬因子(Ihh)、patched-1(Ptch1)、 smoothened(Smo)、神经胶质瘤相关癌基因同源物1(Gli1)的表达。采用蛋白质印迹法评估软骨中基质金属蛋白酶-13(MMP-13)、X型胶原和含血小板反应蛋白基序的解聚素和金属蛋白酶5(ADAMTS-5)的蛋白表达。

结果

组织学分析显示,佐剂诱导的OA组有明显的软骨退变,包括软骨细胞密度降低、软骨变薄和退变,以及细胞外基质中蛋白聚糖含量减少。与相应对照组相比,OA组中Ihh、Ptch1、Smo和Gli1的表达呈时间依赖性升高。与相应对照大鼠相比,OA软骨中MMP-13、X型胶原和ADAMTS-5的蛋白水平显著升高。

结论

这些结果表明,Ihh信号通路的激活可能与佐剂诱导的TMJOA中髁突软骨的病理变化相关。

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