Stromal "activation" markers do not confer pathogenic activity in tendinopathy
Stromal "activation" markers do not confer pathogenic activity in tendinopathy
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DOI：
10.1002/tsm2.204
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发表时间：
2021-03-01
影响因子：
1.2
通讯作者：
Millar, Neal L.
中科院分区：
文献类型：
--
作者：
Crowe, Lindsay A. N.;Melchor, Emma Garcia;Millar, Neal L.
肌腱病是一种非常普遍的肌肉骨骼病理学，与重复性微创伤导致的增量损伤相关。我们试图探索人类肌腱病模型中基质“激活”特征的生理意义。从接受肩部手术的患者中收集撕裂的冈上肌腱和匹配的完整肩胛下肌腱活检，而从接受前交叉韧带（ACL）重建的患者中收集健康肌腱。使用 qRT-PCR 和免疫组织化学在转录物/蛋白质水平上分析基质活化标记物的表达。基质活化标记物的基因表达通过 siRNA 介导的敲低而沉默，或通过 IL-1 β 刺激诱导。在人肌腱组织中鉴定出“激活”标记物平足蛋白、VCAM-1 和 CD248 的表达。 Podoplanin 和 VCAM-1 表达在肌腱病组织中显着增加。正常和肌腱病肌腱细胞中足足蛋白和 VCAM-1 的敲低对基质基因 COL1A1、COL3A1、TNC 或 DCN 的表达没有任何显着影响。同样，在 podoplanin/VCAM-1 敲低培养物中，未观察到炎症介质 IL-6、IL-8 和 CCL2 释放的变化。我们的数据表明，基质“激活”标记物的沉默表达不会影响肌腱细胞的内在炎症特征或基质调节行为。我们认为术语“激活”更合适地反映为肌腱细胞行为的改变，其诱导基质微环境和整体组织结构的变化，而不是通过潜在的任意表型特征进行识别。
Tendinopathy is a highly prevalent musculoskeletal pathology associated with incremental injury as result of repetitive microtrauma. We sought to explore the physiological significance of stromal "activation" signatures in a human model of tendinopathy. Torn supraspinatus tendon and matched intact subscapularis tendon biopsies were collected from patients undergoing shoulder surgery while healthy tendon was collected from patients undergoing anterior cruciate ligament (ACL) reconstruction. Expression of stromal activation markers was analyzed at transcript/protein level using qRT-PCR and immunohistochemistry. Gene expression of stromal activation markers was silenced by siRNA-mediated knockdown or induced by IL-1 beta stimulation. Expression of "activation" markers podoplanin, VCAM-1 and CD248 was identified in human tendon tissue. Podoplanin and VCAM-1 expression were significantly increased in tendinopathic tissue. Knockdown of podoplanin and VCAM-1 in normal and tendinopathic tenocytes did not have any significant effect on expression of matrix genes COL1A1, COL3A1, TNC, or DCN. Similarly, no changes in release of inflammatory mediators IL-6, IL-8, and CCL2 were observed in podoplanin/VCAM-1 knockdown cultures. Our data suggest that silencing expression of stromal "activation" markers does not affect the intrinsic inflammatory profile or matrix regulatory behavior of tenocytes. We propose that the term "activation" is more appropriately reflected by alterations in tenocyte behavior that induce changes in the stromal microenvironment and overall tissue architecture rather than identification through potentially arbitrary phenotypic traits.