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首页> 外文期刊>Osteoarthritis and cartilage >Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage
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Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage

机译：Connexin43 Hemichannels在生物力学刺激的颞下颌关节软骨中介导软骨细胞和基质之间的小分子交换

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摘要

Objective: Connexin (Cx) 43 hemichannels play a role in mechanotransduction. This study was undertaken in order to determine if Cx43 hemichannels were activated in rat temporomandibular joint (TMJ) chondrocytes under mechanical stimulation. Methods: Sprague-Dawley rats were stimulated dental-mechanically. Cx43 expression in rat TMJ cartilage was determined with immunohistochemistry and real-time PCR, and Cx43 hemichannel opening was evaluated by the extra- and intracellular levels of prostaglandin E2 (PGE2). Both primary rat chondrocytes and ATDC5 cells were treated with fluid flow shear stress (FFSS) to induce hemichannel opening. The Cx43 expression level was then determined by real-time PCR or Western blotting, and the extent of Cx43 hemichannel opening was evaluated by measuring both PGE2 release and cellular dye uptake. Results: Cx43 expression and intra- and extracellular PGE2 levels were increased in mechanically-stimulated rat TMJ cartilage compared to the unstimulated control. The FFSS treatment increased Cx43 expression and induced Cx43 hemichannel opening in primary rat chondrocytes and ATDC5 cells indicated by enhanced PGE2 release and dye uptake. Furthermore, the Cx43 hemichannel opening could be blocked by the addition of 18β-glycyrrhetinic acid, a Cx channel inhibitor, Cx43-targeting siRNA, or by withdrawal of FFSS stimulation. The migration of cytosolic Cx43 protein to the plasma membrane in ATDC5 cells was still significant after 8h post 2-h FFSS treatment, and the Cx43 protein level was still high at 48h, which returned to control levels at 72h after treatment. Conclusion: Cx43 hemichannels are activated and mediate small molecule exchange between TMJ chondrocytes and matrix under mechanical stimulation.
机译：目的：Connexin（CX）43 Hemichannels在机械调整中发挥作用。进行该研究，以确定CX43血管笼是否在机械刺激下以大鼠颞下颌关节（TMJ）软骨细胞激活。方法：Sprague-Dawley大鼠牙科机械刺激。大鼠TMJ软骨中的CX43表达用免疫组织化学和实时PCR测定，并通过前列腺素E2（PGE2）的超高和细胞内水平评估CX43血管通道。用流体流动剪切应力（FFS）处理原代大鼠软骨细胞和ATDC5细胞以诱导血管通道开口。然后通过实时PCR或蛋白质印迹测定CX43表达水平，通过测量PGE2释放和细胞染料吸收来评估CX43血管通道开度的程度。结果：与未刺激的对照相比，在机械刺激的大鼠TMJ软骨中增加CX43表达和细胞外PGE2水平。 FFSS治疗增加CX43表达，并诱导CX43在原代大鼠软骨细胞和ATDC5细胞中开口，由增强的PGE2释放和染料吸收表示。此外，CX43血管通道开口可以通过添加18β-甘草酸，Cx通道抑制剂，CX43靶向siRNA或戒断FFSS刺激而阻塞。在2-H FFSS处理8小时后，在ATDC5细胞中迁移细胞溶质CX43蛋白在ATDC5细胞中的血浆膜仍然显着，并且在48h处仍然高48小时，在治疗后72h返回控制水平。结论：在机械刺激下激活CX43血管笼，介导TMJ软骨细胞和基质之间的小分子交换。

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  • 来源
    《Osteoarthritis and cartilage》 |2014年第6期|共9页
  • 作者

    ZhangJ.; ZhangH.Y.; ZhangM.; QiuZ.Y.; WuY.P.; CallawayD.A.; JiangJ.X.; LuL.; JingL.; YangT.; WangM.Q.;

  • 作者单位

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    College of Life Science Shaanxi Normal University Xi'an 710062 China;

    Institute of Orthopaedics Xijing Hospital Fourth Military Medical University 15 Changlexi Road;

    Department of Biochemistry University of Texas Health Science Center 7703 Floyd Curl Drive San;

    Department of Biochemistry University of Texas Health Science Center 7703 Floyd Curl Drive San;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

    State Key Laboratory of Military Stomatology Department of Oral Anatomy and Physiology School of;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 骨科学（运动系疾病、矫形外科学）;
  • 关键词

    Animal model; Chondrocyte; Connexin43; Hemichannel; Mechanical loading; Osteoarthritis;

    机译：动物模型;软骨细胞;Connexin43;Hemichannel;机械载荷;骨关节炎;

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